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1. gt gt gt gt LEG ER o 1 S jalalala al 182555 DIT UN UE EE CHARGE 2 Imm TUBE SENSOR quum a amp PUMP MOTOR MOTOR i amp D d Torr DETECT CARD a E 0 4 i AUTOCLONER POWER AUTOCLONE d CARD MECHANISM d BREED CONNECTION CAMERA POSITION CONTROL MECHANISM e J2 TWO MOTORS AUTOCLONE OPTION P2 KIR3 TOUCH SCREEN TWO POTENTIOMETERS MOTOR Bi DATA TAKER CONTROL CHARGE INTERFACE ettet CARD END i Re FLTEST ALIGN Py 42 LED LAMP Hy e ON SERIAL CONNECTOR D vO V 8 PINHOLE DRIVER UPPER BACKPLANE 9 4 90 GATED SENSOR STREAM LEFT RIGHT ADJ H 256K CAMERA INTERFACE MEMORY CONTROL pi MEMBRANE ITT SWITCHES R PRESSURE lt AMPLIFIER AND ANALOG SIGNAL yo e REGULATOR z PROCESSING CARD PNEUMATIC J3 INTERFACE SHERTH A 2 3 KV lee CAMERA PRESSURE HV POWER 9 INTERFACE J1 m J5 REGULATOR SUPPLY 2 SCOPE CAMERA SWITCH 5 J SCORE SYNG DC POWER
2. ABORT ON FROM SORT Shift Registers ORTU SORT PULSES DELAY CARD Sort info clocks through EXT ABORT ON To Sort Output Card at 1 4 drop rate T judgement Event Detected SS R94 l time Sort Envelop NoSort gt Initiate length info Generate Sort Envelop of Latch and gt e New correct length and Polarity decode Right gt Sort delayed Extend sort Left 10 gt Current info Sort I F and Scaler Card newest l to count ry S B E 1 From I F and Look at the four quarter drops 1 Right 9 Gate Sort Scaler Card behind and ahead of the current event i gt Directi _ 2 Terminate gt Direction Counter Stop STROBE I 801 Left E Value reached Digital Enable sort Crystal unless stop Drive State Restart on count set Phase Generator counter extend and stop value Adder a Reached received sequencer 5 from I F and Scaler 2 To I F and Scaler Card am 2 START Enable Counter X Y Decode Digital Compare g Drop Sort envelop commands Enough drops Counter timer from CPU drops STOP 10 mHz Divide to Xtal Clock 2 desired H Test To LED Card Frequency 5 2 mHz Clock Divide by 5 to give 20 times desired drop frequency Filter AGC Se L 20 CLO
3. SHAPER 7003163A FLOW CELL LASER BEAM S 633 MIRROR BEAM STEERING BEAM TRANSLATOR ASSEMBLY Lasers Refer to Table 2 3 1 and Table 2 3 2 for a summary of lasers including technical specifications PN 4237003 2 3 1 INSTRUMENT DESCRIPTION OPTICAL COMPONENTS Table 2 3 1 Laser Summary Air Cooled Water Cooled Air cooled Laser Argon 190 1305 Enterprise Green HeNe Wavelengths nm 488 633 325 1 351 364 1 351 364 488 543 5 2 457 2 457 351 364 3 465 3 465 4 472 4 472 5 476 5 476 6 488 6 488 7 496 7 496 8 501 8 501 9 514 9 514 Maximum Power 15 10 20 1 200 1 200 150 2 5 mW 2 350 2 350 50 3 150 3 150 4 200 4 200 5 600 5 600 6 1500 6 1500 7 600 7 600 8 400 8 400 9 2000 9 2000 Tips Quartz Quartz Quartz All All All Quartz Dyes FITC APC INDO1 FITC FITC FITC PE PE RD1 CY5 PE RD1 PE RD1 PE RD1 ECD PI PI PI PI Cychrome AO AO AO AO PC5 ECD ECD ECD ECD INDO1 INDO1 INDO1 Thiozole Or Thiozole Or Thiozole Or Thiozole Or 2 3 2 42370036 Table 2 3 2 Laser Technical Specifications INSTRUMENT DESCRIPTION OPTICAL COMPONENTS Laser Specifications Cyonics Output power 15 mW of 488 nm Air Cooled mode purity gt 99 Argon t Beam diameter at 1 e2 0 65 mm Beam divergence 0 95 mrad Polarization ratio Amplitude no
4. 130 INAWNOSISSY dV 93499 V681 00 Wadd 901 INI 901 1NI oxnv oxnv oxnv LXAW 5301193139 8 INVHVd 0119373 V INVHVd peFe dsip sjeubis passeood 104 951049 129 as UA g INVYWd 10 v Wed 12995 sxnv 104 euDis passaooJd esoouo oJ uonisod ensn s yead c u98J9s 8doos uo g WYYYd pallea si 391049 N3349S ead Dot yesHajul ZXNY 101 S391049 NIYIN uonisod jensn FUN U SAV1dSId 9919 9009 uo v WYYYd D si 391049 pue 10 1 LXNY 210 S391049 S3910H9 1VN9IS d3883908d ZXNY ANY LXNV 3998 501140 SAV IdSIG 1140 3998 3905 SAW1dSIG 3998 34095 Wad exnv 3995 Wed TOHLNOD 1091009 SAVIdSIC div 0319 901 L XnV V INVuVd 909 Js3u0mi N9ISSV 1VN3IS 3998 wd NOD 95309115 SAV1dSIG 66 6 1901 GA 10 LINd Z LNd yOu XNy ureeg saddn 10 ZLNd 399108 399108 1 pegay jeddn ggp uofuy yids 7 e Xnv L Xnv LXNY se 9 656 ZLINd Buiubissy 9155 ups cl sue ejeuiuui 3 10 c LW 91099 8 SLINd S 104 yNejeq INVuvd 9410994 Wad v LINd 104 pejoes S INON JI 7 92Jn0S 1 esoouo uay Papae 5914 e
5. This is the FALS OUT FALS HV function J14 P LOG AMP J2 AUX 1 DC RESTORE SEH LOG OUT PROGRAMMABLE J3 DC RESTORE gt GAIN AMPLIFIER CRESTS AUX 1 LINEAR INT OUT J9 PMT2 PEAK 4 e J8 AUX2 PEAK 14 9 J6 AUX2 LOG HH Be P 1 J5 AUX2 INT PMT2 PEAK SEENEN d 1 l J10 OUT MN AUX2 DECODE COMMANDS SET INTEGRATOR SPEED BANDWIDTH FROM BACKPLANE CPU AND CONTROL CARD FUNCTIONS P1 BACKPLANE CONNECTOR 7003041A 2 10 4 PN 4237003C INSTRUMENT DESCRIPTION CARD DESCRIPTION NON GATED AMP NON SWITCH CONFIGURATION EXCEPT ANALYZER 2 11 CARD DESCRIPTION NON GATED AMP NON SWITCH CONFIGURATION EXCEPT ANALYZER Scat CV Amp Card The Scat CV Amp card accepts FALS and 90LS inputs The FALS signal passes through variable attenuator known as the FALS HV Control This signal goes to a variable gain linear amplifier and a log amp See Figure 2 11 1 Figure 2 11 1 Block Diagram Scat CV Amp Card DIGITALLY FALS IN CONTROLLED ji ATTENUATOR INTEGRATOR DC RESTORE This is the FALS HV function LOG AMP ge FALS LOG OUT PROGRAMMABLE Seege S J13 GAIN AMPLIFIER FALS LIN OUT L gt J3 J4 J 5 9015 INTEGRATOR DC RESTORE LOG AMP
6. 7003115 Table 8 2 27 Transformer Drawer Assembly Detail of Parts See Figure 8 2 27 Item Part Number Description 1 6856911 Transformer drawer assembly 2 4506029 K1 and K relay 2 pole 120 Vac 3 4508007 and K4 relay 100 140 Vac 10A 3 32 Vdc input 4 5609034 T3 transformer 100 220 240 118 Vac 50 60 Hz 5 5609035 T1 and T2 transformer 100 120 240 115 Vrms PN 4237003 8 2 27 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 28 Computer Box Assembly See Table 8 2 28 4567 8 o Lae eee 7003113A 8 Table 8 2 28 Computer Box Assembly Detail of Parts See Figure 8 2 28 Part Number Description Item Part Number Description 1 6857479 Computer box assembly 8 N A Spare 2 7260017 Power supply 220 W 9 6028212 Cable from slot 4 3 2016416 Hard drive non removable 10 2016390 2 channel RS 232 Serial card 4 2016298 Floppy drive 5 25 in 11 2016413 V RAM VGA card 5 2016328 Floppy drive 3 50 in 12 N A Spare 6 2016424 Disk Controller card 13 6704665 Lister AB card 7 2016378 Memory Expansion card 2 8 2 28 PN 4237003C Figure 8 2 29 Beam Splitter Dichroic See Table 8 2 29 5 7003131A Table 8 2 29 Beam Splitter Dichroic Detail of Parts See Figure 8 2 29 PARTS LISTS IL
7. AlddflS SNIHOLIMS asyl OI Ol uaindwoo NOILVLSHHOM ER 38 imi 2 1201 p 21 S 190 a SJ0j9euuoo2 BREES 1 JOHLNOO uasvl Lo 33S SAHOLIMS SNVYSsWAW 8 E Jojoeuuoo o N33HOS HONOL J9Y4431NI 1 Ova avno ATddNS ATddNS ATddNS Ld n sAemuyjed 10300 e a z SNEILINN 2 5 7 PN 4237003C INSTRUMENT DESCRIPTION CONTROL AND DISPLAY SUBSYSTEMS Sensor 1 0 Card The Sensor I O card Provides an interface between the upper backplane and the Motor Control card and the Autoclone Sorting Option Control card with the Autoclone Sorting Option Has an analog to digital converter that is fed by a four channel multiplexer Two of the channels read voltages that represent sheath and sample pressure from the Pneumatic Interface card The other two channels are connected to the Motor Control card but are not used Contains the Watchdog circuit which is a protective circuit that monitors Multibus CPU card activity The circuit is a timer that must be periodically reset by the Multibus CPU card If the Multibus CPU card fails to reset the time the Watchdog circuit sen
8. J18VN3 AV T3d LINd 39unos 39unos L Xnv N9SISSV INAWNOSISSY dV Q31V9 681600 KE 901 INI 901 INI exnv oxnv oxnv LXnv LXnv SNOLLO313S 8 INVHVd SNOILOFTAS V Wed pefejdsip sjeubis passaoo4d 391049 15919 uaui g 10 v Nvuvd 199 85 Sxnv 104 euDis passaooJd esoouo Uonisod jensn GIN UI 9915 9909 uo g DIE si a2I0u2 N3342S yead 10 ZXNY 101 S391049 NIYIN uonisod jensn yeed FUN Ul SAW IdSId 9919 9009 uo WYYYd si 391049 pue 10 jesGaqu LXNY 20 S391049 S3910H9 IYN9IS 04 5400 4 ZXNY ANY LXNV ERR SNOILdO SAW1dSId 1140 l 9398 ena vdd 09S SAV IdSIQ IERCH 34095 xV3d e xnv 3998 8 Wed 103109 71031409 SAV 1dSIa day 03199 907 L Xnv V Wgd 909 Js3u0mi N9ISSV 1VNBIS N33u9S id NOD 9S30ufr13 SAV IdSIQ 66 6 mE cl sue ejeuiuui 3 10 yead G Wd 81098 8 SLINd S 104 nejeg V WV 9410994 Mad PLINd L 104 pej99 es S INON JI 7 99110 51 00 uay pae 55914 anos XNY pue 19995 OU euin aures aui je eures eui 1104 099109 dvd 8 91IJ SMOJE SIU ureeg 1901 QA 10 you LXNy ureeg 10 180 1 94
9. HELIUM CADMIUM HeNe ARGON LASER 1 LASER 3 LASER 2 99000005 nana ooo oo TOP VIEW 5965010B 3 7 8 4237003C Figure 3 7 8 1305 Mounting WASHER MOUNTING UPRIGHT Ped Ki 0st e MOUNTING AL PLATE SCREW INSTALLATION PROCEDURES OPTIONAL LASER INSTALLATION 1305 LASER NUT MOUNTING UPRIGHT E LASER CABLE MOUNTING BRACKET PLATE 7003211A Figure 3 7 9 Water Cooled Optical Arrangement TOP VIEW HeNe ARGON LASER 2 WATER COOLED LASER 1 ARGON LASER V LASER LASER TARGETS BEAMS PN 4237003C 633 MIRROR 488 MIRROR BEAM UV MIRROR Reflects 633 NM Reflects 488 NM REDUCER Reflects 325 and transmits and transmits TO 365 NM 320 TO 365 NM 320 TO 365 NM PN 3814286 AND 488 NM PN 3814223 PN 3814222 59650098 3 7 9 3 INSTALLATION PROCEDURES OPTIONAL LASER INSTALLATION Figure 3 7 10 Water Cooled Laser Arrangement pa mea RAIL WATER COOLED HeNe ARGON ARGON LASER LASER 2 LASER 1 0009000009 Oooo ooo oo TOP VIEW 5965 08 WARNING Risk of personal injury The HeCd and water cooled Argon laser will be operated in the UV mode which can injure you when exposed Use extreme care to avoid exposure Carefully note the probable beam paths before firing the laser and block the beam
10. 20 40 60 Microsecond Delay 2 slot 7 J1 PMT Gated Amp slot 6 J8 20 40 60 Microsecond Delay 2 slot 7 J4 PMT Gated Amp slot 6 J9 20 40 60 Microsecond Delay 2 slot 7 J7 PMT Gated Amp slot 6 J10 20 40 60 Microsecond Delay 2 slot 7 J10 PMT Gated Amp slot 6 J11 20 40 60 Microsecond Delay 2 slot 7 J3 PMT Gated Amp slot 6 J13 20 40 60 Microsecond Delay 2 slot 7 J6 PMT Gated Amp slot 6 J14 20 40 60 Microsecond Delay 2 slot 7 J9 PMT Gated Amp slot 6 J17 20 40 60 Microsecond Delay 2 slot 7 J12 PMT Gated Amp slot 6 J18 20 40 60 Microsecond Delay 2 slot 8 J1 PMT Gated Amp slot 6 J12 7 Microsecond Delay 2 slot 8 J3 PMT Gated Amp slot 6 J15 7 Microsecond Delay 2 slot 8 J5 PMT Gated Amp slot 6 J16 7 Microsecond Delay 2 slot 8 J7 PMT Gated Amp slot 6 J19 Bundle these cords and continue 7 Microsecond Delay 1 slot 9 J8 20 40 60 Microsecond Delay 1 slot 10 J11 7 Microsecond Delay 1 slot 9 J6 20 40 60 Microsecond Delay 1 slot 10 J8 7 Microsecond Delay 1 slot 9 J4 20 40 60 Microsecond Delay 1 slot 10 J5 7 Microsecond Delay 1 slot 9 J2 20 40 60 Microsecond Delay 1 slot 10 J2 20 40 60 Microsecond Delay 1 slot 10 Scat Aux Gated Amp slot 11 J6 20 40 60 Microsecond Delay 1 slot 10 Scat Aux Gated Amp slot 11 J7 20 40 60 Microsecond Delay 1 slot 10 Scat Aux Gated Amp slot 11 J8 20 40 60 Microsecond Delay 1 slot 10 J10 Scat Aux Gated Amp slot 11 J9
11. AND CONTROL CARD FUNCTIONS P1 BACKPLANE CONNECTOR Peak Scatter Mux SW R Card DIGITALLY J10 CONTROLLED FALS IN ATTENUATOR INTEGRATOR 4 DC RESTORE This is the FALS HV function LOG AMP y 12 FALS LOG OUT pee 71 J4 MUX1 OUT 1 PROGRAMMABLE gt J13 ud GAIN AMPLIFIER DC RESTORE FALE GUT Ly J14 J1 LOG AMP jo 90LS INTEGRATOR _ DC RESTORE T T PMT1 Loo Lj IN PROGRAMMABLE 18 GAIN AMPLIFIER DC RESTORE 5 BE LINEAR OUT oo J8 PMT4 PEAK L e J7 NOT USED e is ee Ge PARAM A OUT J5 AUX1 INT DECODE COMMANDS SET INTEGRATOR SPEED BANDWIDTH FROM BACKPLANE CPU 7003046A The Peak Scatter Mux SW R card accepts input from the Peak FALS detector and processes the signal to provide variable attenuator and programmable step gain control The signal is not integrated See Figure 2 10 4 Additionally this card accepts input from an auxiliary source PMT The input and develops log and gain controlled linear outputs card integrates the 2 10 3 2 INSTRUMENT DESCRIPTION CARD DESCRIPTION ALL CONFIGURATIONS Figure 2 10 4 Block Diagram Peak Scatter Mux SW R Card PADS DIGITALLY J11 7 CONTROLLED PROGRAMMABLE J13 _ DC RESTORE gt m ATTENUATOR C RESTO
12. Card Name Description Minimum Revision Level Pulse Pileup Det TOF New card N A Sort Delay R3 New card N A Sort Oscillator R2 New card N A Peak ADC Existing card B Peak Scatter Sensor Existing card B Note This is on the Scatter Sensor Gated Amp Cont Existing card B Scat Aux Gated Amp Existing card D PMT Gated Amp Existing card D Quad PSH Existing card F Note For any pot or cable problem order the sort sub assembly The panel sub assembly is the front panel with the cutout for the new control panel PN 4237003C 3 13 5 INSTALLATION PROCEDURES SORT PERFORMANCE ESP UPGRADE 3 13 6 PN 4237003 INSTALLATION PROCEDURES 3 ENTERPRISE LASER UPGRADE 3 14 ENTERPRISE LASER UPGRADE Purpose This procedure provides information for attaching and adjusting the optics assembly used with the Coherent Enterprise laser Installation and adjustment information for the laser is contained in the laser manual PN 7232520 6 The laser light exiting the Enterprise laser contains both the 488 nm and the UV 351 nm wavelengths The Enterprise laser is operated in light regulation relative to the UV There is no separate adjustment for the visible output The Elite optical system must separate the two colors provide attenuation for the visible beam provide a means to focus the UV beam and provide beam translation capability The beams from the laser strike the beam splitter which passes the 488nm wavelength and refl
13. 8 2 4 4237003C PARTS LISTS 8 ILLUSTRATED PARTS Figure 8 2 5 Pneumatics Drawer See Table 8 2 5 6 SIDE VIEW 7003093A 4 3 7 6 5 Table 8 2 5 Pneumatics Drawer Detail of Parts See Figure 8 2 5 Item Part Number Description 1 6232490 Sample regulator 2 6232491 Sheath regulator 3 5120109 Drip chamber switch 4 5120109 Waste filter switch 5 5120109 Fluidics pressure switch 6 5120178 Pressure switch 7 6232096 Solenoid valve 8 6211015 Pilot actuator 6232413 Nipple 10 32 9 6857576 Double acting pinch valve 10 6232143 0 3 micron filter PN 4237003C 8 2 5 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 6 Pneumatics Panel Rear View See Table 8 2 6 SS 7003103A Gi Gi Table 8 2 6 Pneumatics Panel Rear View Detail of Parts See Figure 8 2 6 Item Part Number Description 1 6856985 Upper bubble chamber 2 6801536 Check valve 1 3 6856984 Lower bubble chamber 4 6232418 Large PVC barbed fitting 5 6232104 Large metal barbed fitting 6 6232417 Fitting hose barb 0 062 i d PVC 7 1005697 Small metal barbed fitting 8 2306046 Sheath pressure transducer 9 2306051 Transducer mounting bracket 10 6216129 T fitting brass 0 187 i d 11 6214106 Check valve for tubing 0 156 i d 8 2 6 PN 4237003C Figure 8 2 7 Dual CRT and Connector Panel See Table 8 2 7 13 7003114A
14. N DISCRIMINATOR LEVEL 0 5 DISCRIMINATOR SATISFIED ACTIVE LOW 0 TWO CELLS DETECTED ONE CELL DETECTED TIME 10 CELL DETECTION WITH PPU wn e gt DISCRIMINATOR LEVEL DISCRIMINATOR SATISFIED ACTIVE LOW EXTRA PULSE TWO CELLS DETECTED TWO CELLS DETECTED ADDED BY PPU TIME 7003044 The system determines when it has seen a cell by monitoring discriminator crossings pulse in the parameter being used for the discriminator represents one even cell when it passes through the discriminator twice One pass is on the rising edge and the second pass is on the falling edge Cells that are touching or that pass through the laser beam side by side give rise to a single long pulse that does not return to zero between cells In this case the pulse may not dip below the discriminator between cells and the system does not recognize the second cell as a distinct cell PN 4237003C Transistor Bracket Card INSTRUMENT DESCRIPTION SORT SUBSYSTEM The final output power transistors for the crystal drive and sort pulse amplifiers are located on the Transistor Bracket card Note All the connections from the card to the sort hardware are routed through this card Power Supplies Figure 2 8 12 shows the location of the power supplies and Table 2 8 2 defines each power supplys function Table 2 8 2 Power Supply Functions Power Supply Function 3000 V Deflection Power Sup
15. 0181 03199 Ld 91018 555 ET SEE 900 9 nos 0151 H HOLIMS NS UE 1018 5555 lt 5 25 900 9 usb 051 ENS IF v 101S E 5 6 060 9 5 0151 8 4 1018 1018 344 241016 7280 9 051 H 1051109 div 039 1 101S 3 9 11 3 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE 3 9 12 Table 3 9 3 Adjustments 7 Microsecond 20 Microsecond 40 Microsecond 60 Microsecond PMT Slot Delay Adjustment Slot Delay Adjustment Delay Adjustment Delay Adjustment FAS 9 R105 10 R95 R99 R103 9 R107 10 R96 R100 R104 AUX1 9 R112 10 R97 R101 R105 AUX2 9 R116 10 R98 R102 R106 PMT2 8 R105 7 R95 R99 R103 PMT3 8 R107 7 R96 R100 R104 4 8 R112 7 R97 R101 R105 PMT5 8 R116 7 R98 R102 R106 Applies only if 5th PMT is installed PMT1 Adjustment See Table 3 9 3 and Figure 3 9 4 for additional information on adjustments 1 9 At the Cytometers Sort Screen a Turn the Strobe off b Reset the frequency to 32 kHz Open the detector compartment and remove the following filters 525 BP 575 610 675 640 DL At the Scope Screen set the traces a Put PMT2 INT on Trace 1 bottom b Put PMT1 INT on Trace 2 top At the Option Screen set Align LED to Pulsed 10096 and 2 kHz At the Main Screen adjust the pulses a Adjust P
16. Joyerauab aye6 ou P aounos 1 815 uau 51 AL uoneururejuoo Jeubis S1v4 Aejap ayy 4 18u980H YIM Ing spem eouQ l dad sjeubis 4 sees L d SIUL feraq mopuM MOPUIM ILIM 12815 jene11e00u a 1 syGuajanem 1904 09421 066 ULY 1 5 pe 0400 dwy Deeg 10 4409 paye Su sassed 19414 21010910 WU 066 14 389 pue 4045 ureaq jeddn amp eme ose A 1929 99 0 H Ka Jo um Ajjensn 105195 1949 uomeururejuo2 4 A 154980H inq 2461 1su290H INIL ugesot sou saos tg im mag samo WN an uasa lt Id Woy 8 soueselon 2 gt 38 1999 101513 01551 9 d 1su2990H c 056 E 1999405 dwy 9 2 4 4 2 6 7 PN 4237003C INSTRUMENT DESCRIPTION ACQUISITION SUBSYSTEM Because of the overlap of the emission bands both PMTs see the emissions from both dyes In most cases the filters and subtraction color compensation cannot compensate for the overlap The Gated Amp solves the problem The following sequence provides a summary of what happens when a single cell passes through the laser beam 1 As the cell passes
17. PN 4237003C Beam Steering Optics When an Elite is configured with multiple lasers beam steering optics are used to allow the beams to be combined or precisely separated at the flow cell Dichroic mirrors pass and or reflect specific wavebands the mirrors from the optical component of the beam steering assemblies Figure 2 3 3 illustrates an alignment mirror 2 3 7 2 INSTRUMENT DESCRIPTION OPTICAL COMPONENTS Figure 2 3 3 Alignment Mirror VERTICAL ROTATION KNOB HORIZONTAL ROTATION KNOB FINE HEIGHT ADJUSTMENT SETS HEU ADJUSTMENT SETSCREW SETSCREW 5965012A Beam Translators A beam translator displaces the laser beam without changing its orientation This is the same effect as moving the laser but the beam translator simplifies the process because the movement is more controlled Beam Expander Reducer beam expander reducer Figure 2 3 4 balances the characteristics of small air cooled lasers with those of large water cooled or HeCd lasers This enables both laser types to be focused at the same distance from the beam shaper Reverse the beam expander reducer if you want to perform the opposite function If the expander reducer is turned so the expander side is used you can reverse it by turning it around to the reducer side 2 3 8 PN 4237003C INSTRUMENT DESCRIPTION OPTICAL COMPONENTS Figure 2 3 4 Beam Expander Reducer VERTICAL TRANSLATION LATERAL TRANSLATION F
18. No Do other valves operate 165 Is the pinch tubing stuck closed 59 or clogged Measure power supply No voltages at Pneumatic Intfc Card connection Replace tubing Are voltages correct No Yes Turn Sheath OFF Is voltage 24V present Yes Remove sheath line from flow cell Repair Power on solenoid Momentarily turn Sheath On Supply Does liguid spout from the disconnected tube No Yes Check cable connections Check Wiring between Replace Solenoid Replace Sheath Unclog or Repllace Pneumatic Solenoid and Pneumatic Filter Replace Flow Intfc Card Infc Card Cell Tip 7003082A PN 4237003CB 7 3 3 TROUBLESHOOTING TROUBLESHOOTING FLUIDIC PROBLEMS 7 3 4 4237003CB 7 4 TROUBLESHOOTING TROUBLESHOOTING PRESSURE CONTROL PROBLEMS TROUBLESHOOTING PRESSURE CONTROL PROBLEMS Purpose Do this procedure to troubleshoot sheath and sample pressures Tools Supplies Needed Pressure gauge Procedure PN 4237003C Sheath Pressure Problems Troubleshooting this system can be difficult because of the feedback from the in line liquid pressure sensor This can be turned off with the jumper on the Pneumatic Interface card to troubleshoot Figure 7 4 1 shows how to troubleshoot for sheath flow Note You should find that little if any adjustment is needed In this configuration the system uses the liquid pressure reading to adjust the sheath air pressure to maintain constant
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20. HOLINIWIEIGIO 399141 930 Bulsn Jaye a Xoq AV 13d Aejap 0 9esn 08 110 30 49019 x GL XVIA 42012 uo Buruado mopuim zo ejoq pefejdsip st uonouny juasaid dwe 31avsiq T18vN3 Bursn 134V HLGIM 0 98sn oc 111334 49019 x 61 ve anjeA 490 9 uo MOpUIA payed Jo 490 2019 UMOUS 202 Juans 2951 OL 2 G 98sn 98sn 31e 1019919 9019 INv38 u3ddn AIS TO 38 L 4399181 8399141 diy 0319 v 193138 501 507 901 901 901 901 901 exnv ixnv PLE end zuwa Lud Sov xv3d vad vaa aad vad exnv ixnv PL end zuwa Lud Sov 9919 OUND payey c g e aJnBiJ PN 4237003C 3 9 14 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE 10sues sseJd 0 suyas Josues 194119 s ejds p uoe3 7 au aures 3U je Nd aui 0011991109 993 SMOJE
21. Enter 5 the number of the Epsonx Prn line and press Enter 6 Press 7 REM and press 8 Press Enter Locate a blank line and note the line number Type the line number noted and press 1 to insert and press Enter Edit the CGI CFG file a Type DRIVER C ELITE GSS HPPCL5 SYS G PRINTER and press Enter b Type HPPCL5 PRN and press Enter c Type RESOLUTION 300 and press Enter d Type COLORS 16 and press Enter e Type FF ON and press Enter PN 4237003C PN 42370036 10 11 INSTALLATION PROCEDURES COLOR PRINTER UPGRADE If FF OFF appears without a REM statement before it add the statement a the number of the FF OFF line and press Enter b Press c REM d Press Verify that the For8 5x and press file is setup correctly for the desired paper size 11 in paper PAPER LETTER is required Add if necessary 1 Locate a blank line and note the line number 2 Type 3 Type the line number noted and press 1 to insert and press Enter PAPER LETTER and press Ente 4 Press cu C L then press Enter b For A4 paper PAPER ISOA4 is required Add if necessary 1 Loca 2 Type 3 Type te a blank line and note the line number the line number noted and press 1 to insert and press Enter PAPER ISOAA and press Enter 4 Press ctri C L then press Enter AT
22. Josuas e asn 0 gp ayy SIU pa 1 Aq pasneo 13661 e se qeua uauM 318VSIQ 318VN3 LN 31v9 dq3snv9 d399lH 1 INV38 H3M01 03201 39v 40 134e 0 uonoy 991049 enjoe mous 9919 9909 uo MOQNIM 319 Z 3ounos 9161 1 0190 Jaye 9 NIV9 OIL abueyo 9129 JO 001 0 03005 K 01 934 93 K 01 9NI 6 5 104145 39095 39V NIV9 5 9399191 NIVIN 140 3345 9919 3402 bid 26 6c L 34098 NILLIS ZEN 93sn 08 1003201 aD 318519 3199 001 0 Aag Wa Y3M07 NIY9 N9ISSV 9NILI3S OWL JASN Sz Wad G3LV9 3401834 HLGIM SI 93sn 09 SL10 01 0 9 1138 Av DO wieasia 0381 5 ERR 053 14 JAYS us emt 2019 IO 7041109 dNY 43199 L 5599044 134e 9 49514 anea 9 1 SHOA 01 0 51 0001 0 YOSIG VEUL 1342316 SI 21 0 496614 UYM uasouo 1 19663 isureDe 01 001 934 93 934 9NI 0L 001
23. aser Head crated Length of 201 cm 79 in Width of 53 cm 21 in Height of 48 cm 19 in Weight of 39 kg 86 Ib Dual CRT Installation If the Elite system arrives without the dual CRT assembly attached perform the following additional steps 1 2 After you move the Cytometer to the installation location and Before you attach the computer pedestal to the table top Verify that the system is disconnected from the main power source Remove the inner access panel under the PMTs ATTENTION It is a good idea to have someone assist you with sliding the dual CRT assembly onto its post 3 Slide the dual CRT assembly onto its post as shown in Figure 3 2 2 IMPORTANT Risk of misleading results Cables that are caught or pinched can cause misleading results Ensure that the cables are not caught or pinched 4 Ensure that the connecting cables pass through the covers slot into the instrument Note If the upper bearing was pushed out reinstall it before proceeding Attach the screw and cap to the top of the post as shown in Figure 3 2 2 Install the limit screw and plastic washer at the base of the post as shown Install the cable connectors as shown Be careful not to overtighten the mounting screws PN 4237003C INSTALLATION PROCEDURES 3 INITIAL SETUP IMPORTANT Risk of misleading results A pinched sample line or fiber optic cable can cause misleading results Use care to avoid pinchin
24. 36V 90V FROM SORT OSCILLATOR CRYSTAL DRIVE ADJUST AMPLITUDE m 36V 90V TRANSISTOR HIGH VOLTAGE BRACKET AMPLIFIER 36V 90V EE FROM SORT OSCILLATOR RS FF po DECODE RS COMMANDS FF L ms FF STROBE FROM SORT OSCILLATOR FL TEST FREQUENCY JI ES Es TT 36V 90V see schematic for other control functions HV ON COMMAND TO 2000 3000 V SUPPLY 5 STROBE DRIVE TO LEDS CPU CONTROLLED CURRENT SOURCE 0 100 FL TEST T DRIVE TO FL TEST LED VFCK FROM SORT OSCILLATOR 7003052A 2 8 14 PN 4237003C INSTRUMENT DESCRIPTION 2 SORT SUBSYSTEM Pulse Pileup Det TOF Card The Pulse Pileup Det TOF card card Figure 2 8 9 performs two major functions TOF time of flight provides an output voltage proportional to the length duration of the input pulse This function applies to particle sizing and doublet detection See TOF Function for additional information detects pulses cells or particles that occur closer together than can be detected with current techniques This function helps improve sort purity by aborting events where contamination could otherwise occur See PPU Function for additional information Note When PPU is enabled the PPU parameter 1 generates a positive PPU detect if tha
25. Do not leave the laser unattended if there is a chance than an unauthorized person may attempt to use it Radiation Hazards WARNING Risk of radiation exposure To reduce the risk of exposure to radiation do not use controls or adjustments or perform any procedures other than those specified in this manual In the design and manufacture of the Elite flow cytometer Beckman Coulter has complied with the requirements governing the use and application of a laser as stipulated in regulatory documents issued by the U S Department of Health and Human Services and by the National Center for Devices and Radiological Health CDRH In compliance with these regulatory PN 4237003C PN 42370036 INTRODUCTION SAFETY PRECAUTIONS documents every measure has been taken to ensure the health and safety of users laboratory personnel and service personnel from the possible dangers of laser use The laser is classified as Class I when it is in the system with the protective housing in place CDRH approved labels are placed near or on those covers that when removed might expose laser radiation See Figures 1 2 1 through 1 2 10 for the labels and their locations on the lasers Figure 1 2 1 Laser Warning Labels Coherent Innova 300 Water Cooled Argon Laser Figure 1 2 2 Laser Warning Labels Innova 90 Water Cooled Argon Laser Figure 1 2 3 Laser Warning Labels Helium Cadmium Laser Figure 1 2 4 Laser Warning Labels Omnichrome Helium Cadm
26. 6 7 1 i 3 7003148A Table 8 2 24 Autoclone Sorting Option Beak Waste Catcher Detail of Parts See Figure 8 2 24 Item Part Number Description 1 1022047 Cylinder holder 2 1022048 Shaft Autoclone Sorting Option beak 3 1022049 Plate Autoclone Sorting Option beak 4 6858880 Autoclone Sorting Option beak 5 1018230 Beak actuator bellows 6 1007908 Threaded pin 7 6232037 Air cylinder single acting 8 2 24 4237003 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 25 Autoclone Sorting Option Location and Connections See Table 8 2 25 O f mmen emm 2 7003147 Table 8 2 25 Autoclone Sorting Option Location and Connections Detail of Parts See Figure 8 2 25 Item Part Number Description 1 6028268 Main cable 2 6705191 Autoclone Sorting Option card 3 6704313 Autoclone Sorting Option mechanism PN 4237003C 8 2 25 8 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 26 Autoclone Sorting Option Accessories See Table 8 2 26 7003143A Table 8 2 26 Autoclone Sorting Option Accessories Detail of Parts See Figure 8 2 26 Item Part Number Description 1 6856635 Cover bezel 2 5415377 Modified socket 3 6704311 Deflection plate 8 2 26 PN 4237003C PARTS LISTS 8 ILLUSTRATED PARTS Figure 8 2 27 Transformer Drawer Assembly See Table 8 2 27 1
27. Determine the revision level of the Dual Laser Controller card in the Elite Ifthe revision level is not D or later order a replacement card immediately If the revision level is D or later go to step 5 Remove all covers from the optical bench and laser areas Set the end panel in place temporarily Note The water cooled laser does fit through the padded opening Using Figure 3 7 1 as a guide a Route the interlock cable attached to the panel down to the Multibus area b Attach the relay bracket to the Multibus card cage Wire all the connections as shown in Figure 3 7 2 PN 4237003C INSTALLATION PROCEDURES 3 OPTIONAL LASER INSTALLATION Figure 3 7 1 Path of Interlock Cable to Multibus System 1 LEFT FRONT RIGHT CORNER OF CYTOMETER FRONT VIEW OF LEFT SIDE OF PEDESTAL 7003014 WITH COVER REMOVED PN 4237003C 3 7 3 INSTALLATION PROCEDURES OPTIONAL LASER INSTALLATION Figure 3 7 2 Cable Connections for Laser Installation These are the connections on the end panel The external interlock cable to the particular laser installed are connected here THIS CABLE IS PART OF THE END PANEL ASSEMBLY SERGE RELAY PANEL 4 PIN MATE N LOCK RED 1 mu BLACK 2 mu GREEN gi WHITE 4 a 2 PIN MATE N LOCK YELLOW WIRE TO PIN 18 BLACK WIRE TO PIN 20 Insert these pins into the existing plug on the laser control card on the J4 connector EXISTING CONNECTIONS FOR HEN
28. xvi TOLERANCES AND LIMITS A 1 1 Pressure 1 1 Compressor Module A 1 1 House Air 1 1 Pneumatics 1 1 Switches A 1 1 PN 4237003C PN 42370036 2 Initial Installation Settings 1 1 Argon Laser A 1 1 Camera 1 1 Front and Back Porch Settings for Best Streams A 1 1 Test Points A 1 2 Pneumatic Interface Card A 1 2 Power Supplies A 1 2 Optics A 1 2 Beam Expander A 1 2 Signal Adjustments A 1 2 Beam Translator Upgrade A 1 2 Gated Amp Laser Alignment Time Space Separated Laser Alignment A 1 3 JUMPERS AND SWITCH SETTINGS 2 1 Jumpers for Circuit Cards Used During Installation and Upgrade Procedures A 2 1 3 PMT Sub SW R2 Card A 2 1 HV DAC Card A 2 1 HV DAC Control Card A 2 1 HV DAC Control Card Gated Amp 5 PMT option A 2 1 Lister AT Card on PC Model 486 A 2 1 Mux and Scope Card A 2 1 Sensor Interface Card A 2 1 Jumpers for Cards in Card Cages A 2 2 Multibus Card Cage A 2 2 Non Gated Amp Card Cage A 2 2 Gated Amp Card Cage A 2 3 Data Acquisition Card Cage A 2 3 Jumpers for Line Voltages A 2 4 OPTICAL SETTINGS A 3 1 Collection Optics Upgrade A 3 1 Sort Waveform Verification A 3 1 Crystal Drive Test Procedure A 3 1 Crystal Drive Adjustment Procedure A 3 1 Sort Pulse Amplitude Test A 3 1 Power Supplies A 3 2 FILE PATHS B 1 1 1 LOCATION OF SOFTWARE FILES B 1 1 ABBREVIATIONS ABBREVIATIONS 1 INDEX INDEX 1 TRADEMARKS xvi
29. 7 5 3 7 TROUBLESHOOTING TROUBLESHOOTING THE SORT SUBSYSTEM Table 7 5 1 Troubleshooting Sort Problems Continued Condition Symptom Problem Corrective Action Low frequency Frequency is Pinched sheath tubing Follow sheath tubing from the flow body to required to normally around release the constriction establish 30 kHz but now d Note Tugging gently on the sheath tubing at the breakoff gives high breakoff point before it disappears behind the sort below 20 kHz collection area may alleviate the problem Clogged sheath filter Change the sheath filter Sheath pressure too low Increase sheath pressure to 12 psi Large orifice 140 um flow Verify that the correct tip is installed cell tip installed Low crystal Crystal drive is Small orifice 76 flow Verify that the correct tip is installed drive required usually 60 75 cell tip installed to establish now only 10 or breakoff lassie needed to Clog in flow cell Remove and clean the flow cell Inspect before obtain a high droplet breakoff point replacing Sort streams waver Sort streams waver from side to side with all three streams in sync Sort stream deflection oscillates sometimes it is too narrow other times it is okay Sort collection area door is open or there is no filter in the biohazard filter slot behind the sort collection area Close the sort door and ensure that there is a biohazard filter in th
30. Condition Symptom Problem Corrective Action No droplet Adjusted crystal HV Deflection plates arced 1 Turn off Cytometer save settings first if breakoff drive frequency did causing crystal drive circuit desired not respond to lockup 2 Clean and dry the deflection plate and power system on Blown fuse on Sort Output card Replace the fuse Power supply problem Check the power supply Mist in sort area Mist appears in sort area Wet plates Liquid buildup on top of sort tubes Poor sort purity Satellite drops downstream have not recombined 1 Adjust crystal drive frequency while looking at drops downstream to ensure that no satellite droplets Figure 7 5 2 are present 2 Dry the plates Fanning between side and center streams Constant fan of side streams into center stream Wet plates Liquid buildup on top of sort tubes Poor sort purity and recovery Filament connecting last attached drop is too thick or too thin Adjust crystal drive setting to tighten the side streams Data rate is too high for particular sample type 1 Lower the data rate 2 Filter the sample with nylon mesh Dry the plates Incorrect phase setting Change the phase setting Irregular fanning of side streams into center stream Valves regulators sound like they are constantly cycling clicking Sheath pressure is not constant 1 Find leak in
31. PN 4237003C 3 7 15 INSTALLATION PROCEDURES OPTIONAL LASER INSTALLATION 3 7 16 4237003 INSTALLATION PROCEDURES HOUSE AIR INSTALLATION 3 8 HOUSE AIR INSTALLATION Procedure 1 2 3 4 10 11 12 13 14 PN 4237003C Turn the instrument off Open the pneumatic housing door Remove the four screws securing the bottom side cover and remove the cover Open the bottle tray door and disconnect unit system air quick connect QD22 blue and system vacuum QD23 yellow from compressor Disconnect the white connector not used with this kit Detach the compressor from support shipping brackets if attached and remove compressor from the pneumatic housing Remove the Compressor shipping bracket located below valve bracket Attach gauge and regulator box inside pneumatic housing at the compressor shipping bracket location Note Use the same shipping bracket screws for the installation or the new screws provided Ensure the box system pressure regulator knob is rotated fully counterclockwise Connect QD22 blue and QD23 yellow into corresponding box connectors system air and system vacuum Connect orange and white quick connects to the same color body couplings of the box called house air and house vacuum Attach the two air and vacuum lines to the customer air and vacuum system to be used Split into two lines vacuum and air to suit customers needs Turn unit power ON Ensure the system
32. Quad Peak Sense and Hold PSH Card input The end of DISCSAT triggers a timer DISCSAT to begin the Discsat Extension time selected This line is low while by the user At the end of the extension the the input pulse exceeds gt CLOSE signal is set high This blocks all the the discriminator level PSH circuits from accepting any more input The start of DISCSAT causes the ADC card to set the lt OPEN line to low This enables the PSH cards to charge to 4 input signals peak value Ke 23 y When low the PSH capacitor D charges and holds the voltage La When high the capacitor is Ee SE Es shorted and the stored voltage Pulses are 74 S4 is erased accepted in this Jk 4 SCH The stored values are applied to the time period NS ADC integrated circuit one at a time CLOSE M eo x and converted to digital values These When low the circuit converted values are placed on the allows the pulse to enter Datapath At the end of this process When high the input is grounded gt Sua determined by the number of parameters 10 prevent any input SATENSION Oft OPON sereen being converted the OPEN line goes high to reset the PSH capacitor and the CLOSE line goes low to enable the PSH card to accept a new input Quad Peak Sense and Hold PSH Card Output Stretched Pulse 7003072A 2 13 6 PN 4237003C SIGNAL SELECTION AND DIGITIZATION Figure 2 13
33. V6L0 007 NOILLVIQVH 9345311995 HO LOAYIG OL SHNSOdxA NIMS 3A3 GIOAV 95343334 MOOTHALNI N3dO NAHM NOILLVIQVH H3SV1 3 TI8ISIANI HO QNV NISIA 1 SIHL SI NOLLVIGVH H3SV 1 3 I8ISIANI HO GNV 3 18ISIA 3unsodxa 193330 90 1H31N gay Hulsuas sjaqe 18se 01 2 1 941614 PN 4237003C 1 2 8 INTRODUCTION SAFETY PRECAUTIONS Notify the Customer About Barrier Protection Bring the following warning to the customers attention before advising the customer to perform any service maintenance troubleshooting or service procedure on the instrument WARNING Risk of personal injury or contamination If you do not properly shield yourself while performing service maintenance and troubleshooting procedures residual fluids in the instrument could injure or contaminate you Beckman Coulter recommends that you wear barrier protection such as appropriate safety glasses lab coat and gloves throughout the performance of service maintenance and troubleshooting procedures to avoid contact with cleaners and or residual fluids in the instrument PN 4237003C 1 2 9 INTRODUCTION SAFETY PRECAUTIONS 1 2 10 PN 4237003 CONTENTS 2 INSTRUMENT DESCRIPTION 2 1 1 2 1 INSTRUMENT OVERVIEW 2 1 1 Function 2 1 1 Description 2 1 1 Main Components 2 1 1 Subsystems 2 1 1 P
34. a Loosen the setscrew that secures the camera to the horizontal tube and slightly move the camera Note This step requires you to readjust the primary focus as instructed in step 2 Make small adjustments until optimal zoom focus is achieved Note Optimize the focus at a high zoom setting so the droplets can be clearly seen for sorting Minimal loss of focus at extreme zoom settings is acceptable Zooming the Camera 1 Zoom out as far as possible to get the smallest image 2 Verify that the zoom number in the periscope window is 1 adjust if necessary Note Older cameras with the hose clamp secured periscope may go to 0 7 which is acceptable ATTENTION Do not lose the screws Do not let the belt fall off 3 Adjust the zoom a b i OR Loosen the screws shown in Figure 4 1 1 to release the tension from the pulley Without rotating the pulley rotate the zoom to the 1 or 0 7 position Tighten or replace the belt and the screws Carefully loosen the setscrew that secures the belt pulley to the feedback pot Use a short screwdriver to move the pot shaft inside the pulley Note The pot motor and control card form a closed loop Therefore if you move the pot the motor runs continuously to attempt to re zero the pot Do not allow the motor to run past the limits of the zoom mechanism The proper adjustment technique is to turn the pot shaft slightly to let the motor move in small increments Tighten the set
35. c Set Drop Sorted to 1 Verify operation of abort for two events in one drop On the Cytometer Sort screen press the Coincidence Abort block This block should cycle through three states as you press it Coinc Coinc Coinc Abort Abort Abort Off On PPU Det On Press Coincidence Abort until it is in the Coinc Abort Off state Note Non ESP systems will not display the PPU coincidence option Adjust R109 on the Pulse Generator card to separate the pulses by about 80 microseconds Note At this time the sort count should be counting and the sort rate should be about the same as the data rate Connect your oscilloscope to the alligator clip on the sample insertion rod Adjust Defection to 1596 and adjust your scope to observe the sort pulses Press Coincidence once to switch to the Coincidence On state Note ESP systems can be set to either Ext Sort or Comp Abort Slowly turn R109 to reduce the time between the two pulses Note At about 30 microseconds separation between the pulse rising edges you will see the coincidence counts increase while the sort rate decreases This verifies operation of the abort within one drop function Pulse Pileup Verification This procedure applies to ESP equipped systems only 1 Continue decreasing the pulse separation until the sort rate increases again This occurs as the two pulses begin to blend and an a discriminator crossing does not occur to separate the pulses Press Coincidence Abor
36. p 7003091 8 Table 8 2 1 Elite Cytometer Right Panel Removed Detail of Parts See Figure 8 2 1 Item Part Number Description Item Part Number Description 1 6704753 Camera inside 7 N A Data Acquisition card cage 2 6602642 Flow cell inside 8 6856911 Transformer drawer 3 6856842 Sample collect drawer 9 6856950 Connector panel 4 6856971 Sample station 10 6856505 PMT 4 inside 6857203 Door 5 6856970 Mixer motor assembly 11 6856833 PMT 5 inside 6 N A Gated Amp card cage 12 6856942 PMT plate inside PN 4237003 8 2 1 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 2 Workstation See Table 8 2 2 7003121A Table 8 2 2 Workstation Detail of Parts See Figure 8 2 2 Item Part Number Description 1 2016161 Mouse 2 2016670 Computer Pentium Workstation pc 3 6856936 PC Pedestal 4 2016592 Keyboard 5 2016651 Monitor color 17 in N A 1411141 Chair N A 6914969 Printer color 8 2 2 PN 4237003C PARTS LISTS ILLUSTRATED PARTS Figure 8 2 3 Sheath and Waste Compartment See Table 8 2 3 ET 7 17 Pe 4 15 a a _ lemseresl 7 7003097A Table 8 2 3 Sheath and Waste Compartment Detail of Parts See Figure 8 2 3 Item Part Number Description Part Number Description 1 6856509 Bottle tray door 11 623244
37. 20 40 60 Microsecond Delay 1 slot 10 Scat Aux Gated Amp slot 11 J11 20 40 60 Microsecond Delay 1 slot 10 Scat Aux Gated Amp slot 11 J12 20 40 60 Microsecond Delay 1 slot 10 Scat Aux Gated Amp slot 11 J15 IJ J4 J7 J3 J6 J9 20 40 60 Microsecond Delay 1 slot 10 J12 Scat Aux Gated Amp slot 11 J16 7 Microsecond Delay 1 slot 9 J1 Scat Aux Gated Amp slot 11 J10 7 Microsecond Delay 1 slot 9 J3 Scat Aux Gated Amp slot 11 J13 7 Microsecond Delay 1 slot 9 J5 Scat Aux Gated Amp slot 11 J14 slot 9 slot 9 slot 9 slot 9 7 Microsecond Delay 1 slot 9 J7 Scat Aux Gated Amp slot 11 J17 4 10 7 4 SERVICE AND REPAIR PROCEDURES ELECTRONIC ADJUSTMENTS CABLE AND JUMPER CONFIGURATIONS 4 10 8 Table 4 10 6 Cable Connections Continued From To PMT Gated Amp slot 6 J20 3 PMT Sub SW R1 slot 5 J3 PMT Gated Amp slot 6 J21 3 PMT Sub SW R1 slot 5 J2 Gated Amp slot 6 J22 3 PMT Sub SW R1 slot 5 J1 PMT Gated Amp slot 6 J23 3 PMT Sub SW R2 slot 4 J1 PMT Gated Amp slot 6 J24 3 PMT Sub SW R2 slot 4 J2 PMT Gated Amp slot 6 J25 3 PMT Sub SW R2 slot 4 J3 3 PMT Sub SW R1 slot 5 J5 Dual FL slot 3 J1 3 PMT Sub SW R2 slot 4 J5 Dual FL slot 3 J5 Scat Aux Gated Amp slot 11 J18 Peak Scatter Mux slo
38. 25 SE 22 E S lt L 0Vd AH 6 HO E gir Od elid 8 HO lt Ir gp 5011 NEZ 100 2 ee eee s i FH TA gr 14 9 D 1nozu 77 3 Gir gr or Im e NOTH SHI vir gored lt pr 901 1714 H Fop INFZWd emt IN HA Yd p Ld 901 74 IHI Um MEIN EH HH V oer S0TFIWd m lNFFIWd 96 617 x Cu 901 5106 0015106 1NI 106 2a swal Uu 1NI S106 901 51 NI T D ID SB Bak DUT 901 8703 A9 1V9S z l amp Lamm 0 HO 6 INIST 1 Si O 3405 2 IN31SAS IM pue xm OF A AH XV02 VNSIS omg 1925 yead jnouy sdwy ajqeuayms uoN 8 9 6 950500 L L01S 1015 399 9399 1838 Y3ddN 42370036 2 6 10 INSTRUMENT DESCRIPTION 2 ACQUISITION SUBSYSTEM 4 Switchable Amps with Peak Scatter Function The 4PMT Switchable Amps with Peak Scatter Function Figure 2 6 9 was a factory configuration available beginning February 1992 With this setup e fluorescence PMT signals go to the PMT Sub SW R card which compensates all three signals FL1 FL2 and FL3 e FLI and FL2 are routed to the Dua
39. FL3 LOG FL1 OUT FL2 OUT SWITCH R 2 JUMPER AS BD 1 INT 2 J15 AUX 2 PMT2 3 4 5 LIN LOG OR PK J24 FALS PEAK J9 SLOT 14 SCAT CV SWITCH R 1 DUAL MUX 90LS INT PMT1 INT Jii FALS INT N C Ms SEALS INT LOG J12 FALS IN FALS LOG 90LS IN Amour FALSINT Ju DOSEN OR LOG INT INA DUAL MUX N C ui axi Ja E1 E2 0UT G OR PMTA PK PEAK 2 INT J13 PMT2 LOG J14 SLOT 43 NG DUAL FL FLI PK PEAK J18 SWITCH R J5 PMTS J FL2 PK PMT3 INT J18 pidas PMT3 LOG J17 ii PMT4 INT J19 D PMT4 5 cite PMT4 LOG J20 E1 2 OUT 5 INT J22 LOG J23 FL3 LOG FL1 OUT FL3 PK E1 E2 OUT E3 E4 OUT E5 E6 OUT NOT AVAILBLE TRIGGER SOURCE OR GATE WINDOW J25 SYSTEM SOURCE GATE WINDOW J27 PEAK 5 PK PN 4237003C LOWER CARD CAGE SLOT 7 QUAD PSH 1 JUMPER E2 E4 LOWER CARD CAGE SLOT 48 QUAD PSH 2 JUMPER E2 E4 SLOT 9 ADC PSH CONTROL LOWER CARD CAGE 7003029A INSTRUMENT DESCRIPTION 26 19 INSTRUMENT DESCRIPTION 2 6 20 2 6 20 PN 4237003C INSTRUMENT DESCRIPTION OPTICAL ELEMENTS 2 1 OPTICAL ELEMENTS Overview This section describes optical principles as they relate to the optical components used in the Elite system The different aspec
40. Monitoring the Sort 1 4 2 8 Zoom in on the last attached droplet so that both the droplet position and connecting neck are clearly visible Place two cursors around the last attached droplet to monitor the stream stability a Place one cursor tangent to the bottom of the last attached drop b Place the other cursor tangent to the bottom of the second to the last attached drop Occasionally view the side streams while sorting to ensure they remain clean and stable Watch the droplet in the camera while adjusting the drive to ensure that the connection between the last drop and the second to last remains in tact The connection is optimal when the filament between two drops is very thin yet still connected At this point the side streams should be at their best Ifthe side streams appear to fan slightly make minor adjustments to the crystal drive to improve the streams without affecting the sort PN 4237003C PN 42370036 SERVICE AND REPAIR PROCEDURES SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE Ifthe filament connecting the last attached drop breaks decrease the Crystal Drive If the filament fattens increase the drive while viewing the side streams Ifthe side streams do not remain stable stop the sort and investigate 5 Press DEBUBBLE and or CLEAR two or three times Ifthe drop returns to the same place the sort should be fine Ifthe drop does not return to the same droplet investigate furthe
41. Note For the part numbers and location of these schematic files see Chapter 6 Prism and Window Test card Bitmap and Sort Decision card Interface and Scaler card Sort Delay card Sort Oscillator card Sort Output card Pulse Pileup Det TOF card Prism and Sort Window Test Card If rectilinear sort gates are used the sort process begins at the Prism and Sort Window Test card Figure 2 8 3 which Stores the channel numbers that define the upper and lower limits for the sort regions being used The storage occurs before the sort begins when the Sort Settings are sent from the Workstation Compares the values that the ADC presents on the datapath bus to the appropriate stored value matching parameter during the actual sort The sort condition for a particular window and particular parameter is met when current value gt low channel number current value lt high channel number Sends the indication to the Bitmap and Sort Decision card if the sort condition is met Current channel values are compared to the prism level for each parameter via a similar process 4237003C PN 42370036 Figure 2 8 3 Block Diagram Prism and Sort Window Test Card DATAPATH FROM ADC amp PSH CONTROL Regions to store REGISTERS FOR LEVELS 16 10 BIT WORDS L SStrobe PRISM POSITIVE region select CONTROL
42. PN 42370036 The Generator is now ready to use Amplitude of the test pulses is determined by the value written to port 0204 FFFF will give the largest pulses and 0000 the smallest Pulse width is determined by the four jumpers X1 X2 X3 X4 With the jumpers in the pulses are suitable for testing the electronics in the sense in quartz low bandwidth configuration With the jumpers out the pulses are suitable for testing the sense in air configuration 4 2 15 4 SERVICE AND REPAIR PROCEDURES SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE 4 2 16 PN 4237003C 4 3 PN 4237003C LINE VOLTAGE SELECTION SERVICE AND REPAIR PROCEDURES 4 LINE VOLTAGE SELECTION WARNING Risk of lethal electrical shock Disconnect all power cords including the instrument power cord and the air cooled laser power cord from the rear of the unit to prevent electrical shock Verify that the correct line voltage jumpers have been installed according to Table 4 3 1 Table 4 3 1 Line Voltage Jumpers Location Jumpers 100 Vac input to main system transformer TB9 2B to TB9 4B jumper cable TB9 7B to TB9 9B jumper cable TB9 1B to TB9 2B jumper brass TB9 6B to TB9 7B jumper brass BL eer 115 Vac input to main system transformer TB9 2B to TB9 4B jumper cable Sa TB9 1B to TB9 2B jumper brass TB9 5B to TB9 6B jumper brass 230 Vac input to main system transformer TB9 1B to
43. Stream is deflected or unstable Partially clogged flow cell Press SHEATH and VACUUM If still clogged brush tip with a camel hair brush and press SHEATH and VACUUM If still clogged remove flow cell tip and force liquid or air through tip Low data rate Partially clogged flow cell Press SHEATH and VACUUM If still clogged brush tip with a camel hair brush and press SHEATH and VACUUM If still clogged remove flow cell tip and force liquid or air through it Bubbles Press DEBUBBLE Tube not properly sealed Push cap into tube Pinched sample line Work the tubing until pinch is removed Low sample pressure Slightly increase flow rate 4237003C TROUBLESHOOTING ISOLATING THE PROBLEM AREA Table 7 1 2 Autoclone Sorting Option General Troubleshooting Information Continued High data rate Sheath tank not properly sealed Reseal sheath tank Discriminator too low Set to higher value Extraneous light entering sensor Check mask and alignment check for external light sources Long pulse rise time Bubbles or debris in flow cell tip Press DEBUBBLE OR Press SHEATH and VACUUM then press DEBUBBLE Alignment check Shifted mean channel or high CVs Same as all probable causes listed above Press DEBUBBLE OR Press SHEATH and VACUUM then press DEBUBBLE Horizontal beam flow cell and or vertical beam not
44. a Thecolor coded connectors b Thelevel detector wires to the bracket 10 Hold the bottle cap in its normal orientation and verify there is no Waste Bottle Full message on the Cytometer screen 11 Turn the bottle cap over and verify a That the float moves freely b That Waste Bottle Full appears on the Cytometer screen 12 Return to step 8 in Installation Procedure above 3 19 4 PN 4237003 INSTALLATION PROCEDURES COLOR PRINTER UPGRADE 3 20 COLOR PRINTER UPGRADE Purpose Do this procedure to install the Hewlett Packard9 1600C Color Printer which replaces the 1200C Color Printer Instrument model numbers affected are e 2356 Epics Elite ESP with Gated Amp and or Autoclone Sorting Option and AL e 2357 Epics Elite ESP with Gated Amp and or Autoclone Sorting Option 2358 Epics Elite ESP e 2360 Epics Elite with Gated Amp and or Autoclone Sorting Option e 2362 Epics Elite 2819 Epics Elite Analyzer Tools Supplies Needed HP 1600C Color Printer kit PN 6914968 8 which includes gt HP 1600C Color Printer PN 2016675 4 gt Parallel printer cable PN 6027510 6 Elite Color Printer kit PN 6914969 3 which includes gt 1600C Color Printer kit PN 6914968 5 gt Software driver for HP 1600C Color Printer PN 6417268 9 gt Installation Guide for Elite Color Printer PN 4237252 8 HP 1600C Software Driver kit PN 6941966 6 for International use only which includes gt Software driver for HP 1
45. a Zoom in enough to see that the drops are crisp and in focus b Look above and below the ground plate 4 11 5 4 SERVICE AND REPAIR PROCEDURES SORT SENSE FLOW CELL ADJUSTMENT AND VERIFICATION 4 11 6 16 1T Scroll through the Drive Frequency to find the best possible frequency that produces crisp clean droplets and a high enough droplet breakoff so that it is slightly above the ground plate If the drops look crisp and focused but the last attached drop is not above the ground plate either move the plates slightly to see the last droplet or increase the Crystal Drive slightly to achieve the same thing Note Avoid increasing the Crystal Drive above 9096 It is not necessary to have three free drops above the ground plate One drop is sufficient and will not affect your sort in any way As long as the breakoff between the last attached drop and the first free drop is visible the sort will be easy to monitor Ensure that the filament which connects the last attached drop to the one above it is thin it should be thin enough to be connected but not broken off See Figure 4 11 3 Figure 4 11 3 Filament Connecting Last Attached Drop CURSOR PLACEMENT FOR MONITORING LAST ATTACHED DROPLET e eat N THIN FILAMENT BETWEEN LAST DOWNSTREAM ATTACHED DROP AND STREAM SATELLITE DROPLETS RECOMBINING USE THE CRYSTAL DRIVE ADJUSTMENT 10 TO KEEP THIS THIN 7256008A 18 When the drops look sharp crisp and in focus all t
46. e or pmt trigger to discriminator level i Source Ka 9 mm PEN i Select gate or trigger to display LOWER BEAM LOCKOUT When enabled a new gate pulse GATE PULSE ENDED cannot be started until le the current pulse is ended When enabled the gate pulse goes to ENABLE GATED AMP the Mux and scope DISCRIMINATOR card where it is routed OVERRIDE to the DISCSAT line WINDOW WIDTH SHIFT REGISTER lt gt RESET Fy WINDOW DELAY Gate pulse to Scat Aux RENE Gated Amp and SET RESET 4 gt PMT Gated Amp FLIP FLOP cards P1 51 52 SET L GATE 57 ENABLED Two variable length shift registers are used to generate the gate pulse One determines the gate width the other determined the gate delay EE 10 MHZ 10 MICROSECONDS CLOCK 5 MICROSECONDS e SHIFT REGISTER CLOCK 1 MICROSECONDS 0 5 MICROSECONDS PN 4237003C 7003030A 2 12 1 2 INSTRUMENT DESCRIPTION CARD DESCRIPTION GATED AMPLIFIER CONFIGURATION 2 12 2 Gated Amp R3 Card Gated Amp configurations require the Gated Amp Control R3 card Functions The Gated Amp Control R3 card Selects the trigger source between the input from the Scat Aux Gated Amp card or input from the PMT Gated Amp card Attenuates the trigger source via a programmable attentuator then passes the trigger through a log amp The amplified or unamplified linear signal is then selected as
47. from left in the top cage f E9 E10 IN and install in 10th slot from left in the top cage g from left in the top cage h in 12th slot from left in the top cage i slot from left in the top cage from left in the top cage k Thenew Mux and Scope card Table 3 9 1 Gated Amp Jumper Configuration Circuit Card Slot Jumpers Gated Amp Control R 3 1 None Dual FL Switch R 3 E1 E2 OUT 3 PMT Sub W R 2 4 5 PMT E1 E2 OUT E3 E4 IN E5 E6 OUT 4 PMT E1 E2 OUT E3 E4 OUT E5 E6 OUT 3 PMT Sub SW R1 5 E1 E2 OUT E3 E4 OUT E5 E6 OUT PMT Gated Amp 6 E1 E2 OUT Quad 20 40 60 Microsecond Delay 2 7 E1 E2 IN E3 E4 IN E10 E11 IN Quad 7 Microsecond Delay 2 8 E10 E15 IN Quad 7 Microsecond Delay 1 9 E10 E11 IN Quad 20 40 60 Microsecond Delay 1 10 E1 E2 IN E3 E4 IN E9 E10 IN Scat Aux Gated Amp 11 E2 E3 OUT 3 PMT Sub SW R 1 12 E1 E2 IN E3 E4 OUT E5 E6 IN Peak Scatter Mux Sub SW R 13 E1 E2 IN E3 E4 IN Scat CV SW R 1 14 E1 E2 OUT E3 E4 OUT 23 Install the Mux and Scope card in slot 6 of the bottom card cage 24 Connect cards with coax cables as indicated in Table 3 9 2 3 9 4 42370036 PN 42370036 Table 3 9 2 Coax Cable Interconnections INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE From To Quad 7 Microsecond Delay 2 slot 8 Quad 20 40 60 Delay 2 slot 7 J2 J2 J4 J5 J6 J8 J8 J11 Quad 20 40 60 Microsecond Delay 2 slot 7 PMT G
48. long May allow noise or doublet Integral signal has not reached peak to contaminate signal value before window closes The following chart describes the numbers in Figure 2 13 6 Number Description 1 The discriminator level one Peak Sense and Hold PSH channel is exceeded This is called the trigger channel 2 All active PSH channels begin capturing data start of DISCSAT by allowing their hold capacitors to charge to the input voltage level 3 The trigger channels pulse falls below the discriminator level the DISCSAT period ends The discriminator extension DISCSAT EXT period begins 4 The DISCSAT EXT period allows slower signals integral and log time to reach their maximum values 5 The DISCSAT EXT ends The card disables the inputs to the PSH channels the cards retain the voltage that their respective hold capacitors charged to until they can be measured by the analog to digital conversion circuit and reset PN 4237003 2 13 9 INSTRUMENT DESCRIPTION SIGNAL SELECTION AND DIGITIZATION Figure 2 13 6 Peak ADC PSH Card Timing PEAK ADC TIMING STRETCHED PULSE OF INTEGRAL SIGNAL DISCRIMINATOR LEVEL FALS PEAK INTEGRAL SIGNAL ACQUISITION TIMING DISCSAT DISCSAT EXT 4 2 5 5 10 or 15 microseconds 7003038 There are several ways to determine the best ASD You can calculate the time based the following Laser beam spot size Sheath flow velocity e Average cell size
49. 15 Mouse Support Option Disabled Above 1 MB Memory test Enabled Memory Test Tick Sound Enabled Memory Parity Error Check Enabled Hit DEL Message Display Enabled Hard Disk Type 47 RAM Area 0 300 Wait For F1 If Any Error Enabled System Boot Up Num Lock On Floppy Drive Seek At Boot Enabled System Boot Up Sequence A C System Boot Up CPU Speed High External Cache Memory Enabled Internal Cache Memory Enabled Password Checking Option Setup Video ROM Shadow C000 32K Enabled Adapter ROM Shadow C800 32K Disabled Adapter ROM Shadow D000 32K Disabled Adapter ROM Shadow D800 32K Disabled Adapter ROM Shadow E000 32K Disabled Adapter ROM Shadow E800 32K Disabled System BIOS Cacheable Enabled Shadow RAM Write Protection Enabled Boot Sector Virus Protection Disabled 4 8 1 4 SERVICE AND REPAIR PROCEDURES WORKSTATION AND SOFTWARE 4 8 2 10 11 12 13 External Cache Write Mode Wr Back Non Cacheable Block AT Bus Non Cacheable Block Size Disabled Non Cacheable Block Base 512KB 8 16 Bit I O Recovery Time 4 2 CLK Local Bus Ready Transparent Disable Press to return to the Setup menu Select PERIPHERAL MANAGEMENT SETUP Follow the instructions on the screen and ensure the following On Board Floppy Drive Enabled On Board IDE Drive Disabled First Serial Port Address 3f8 H Second Serial Port Address 2f8
50. 17 um ht 34 um width 40 mm front focal length 80 mm rear focal length Table 8 1 3 Cables Part Number Description Figure Part Number 3814178 Light cable 0 125 in diam 60 in long 3814181 Light cable 0 094 in diam 72 in long 6027510 Printer parallel 6027693 Printer serial MDAD System 6027803 Bullet bullet coax cable 27 in 6027937 Data transfer serial 9 pin Cytologic 6027939 Data transfer serial 25 pin Cytologic 6027967 Cable assembly Cytometer 7 5 in long 6027992 KIR 3 communication 41 in long 8 2 7 4 6028002 Cable assembly Cytometer ground 5 in long 8 2 14 4 6028009 Data lister out external 6028016 Laser control cable W49 8 2 35 6 6028018 Charge cable sort transistor W30 8 2 10 4 6028020 Pneumatics sensor motor cable sensor interface 8 2 10 3 6028021 Serial communications cable 8 2 7 14 8 2 10 17 6028022 60 POS bitmap memory ribbon 8 2 10 8 11 6028023 Scope signal cable 8 2 10 9 12 6028024 Pneumatic CPU 8 2 10 13 8 1 2 PN 4237003C MASTER PARTS LIST IN NUMERICAL ORDER CATEGORY Table 8 1 3 Cables Continued PARTS LISTS Part Number Description Figure Part Number 6028025 Data lister out internal 8 2 7 13 8 2 10 7 18 6028026 Data taker PMT interface DAC 6028038 Data acquisition cable 8 2 10 10 14 6028040 36 V cable W19 8 2 10 5 6028045 Dual CRT W35 8 2 7 3 6028067 Serial communication 9 25 pin 6028070 BNC cable assembly W31 8 2 7 19 8 2 1
51. 2 INSTRUMENT DESCRIPTION OPTICAL COMPONENTS Table 2 3 2 Laser Technical Specifications Continued Laser Specifications Coherent DPSSt Wavelength 532 nm Polarization gt 1 100 Output power 20 mW minimum Spatial mode Beam diameter at 1 2 0 6 mm Beam divergence 1 3 mrad Amplitude noise 0 596 RMS 10 Hz 10 MHz Warm up time 5 minutes Beam pointing stability 07 5 urad C Operating voltage 100 115 200 Vac 10 Omnichrome Minimum power when 30 mW HeCd 74t shipped 20 mW Warranted minimum power 325 nm Wavelength Forced air Cooling Multi mode Mode 1 2 mm Beam diameter at ue 0 5 mrad Beam divergence 5 minutes Warm up time from cold 2 minutes Recovery from standby 10 to 40 C Operating temperature gt 6 000 hours Expected tube life 104 128 Vac Line voltage 48 63 Hz Line frequency Optical filters 325 nm beam splitter 325 nm long pass 381 nm band pass 440 nm long pass Coherent Innova Enterprise Wavelength output power Mode Polarization ratio Beam diameter at t e Beam divergence Beam pointing stability Optical noise Power stability Input voltage Input current Cooling water Weight Power supply 488 nm 150 mW 351 364 nm 50 mW 100 1 1 20 mm 488 nm 0 88 mm UV 0 82 mrad 488 nm 0 73 mrad UV lt 5 1 096 RMS 1 096 UV 208 240 Vac 10 50 or 60 Hz single phase 31 A 10 8 16 lite
52. 2 13 1 level of 2 13 1 circuitry how to confirm operation 2 9 4 clogs biofilter See biofilter coincidence verification See sort function verification collection optics fast quartz flow cell 2 3 11 installation 3 15 1 jet in air flow cell 2 3 10 sort sense flow cell 2 3 9 collection options flow cell tips 2 3 9 color compensation function 2 6 1 subtraction circuit operation 2 6 1 subtraction circuit operation illustration 2 6 2 troubleshooting 7 2 3 color printer upgrade See Printers communication pathways from Acquisition card cage to Sensor Interface card 2 9 4 from Multibus CPU card to Pneumatic Interface card 2 9 4 to Pneumatic Interface card from Multibus CPU card 2 9 4 Compressor module air water separator function of 2 9 1 block diagram 2 9 12 connections 2 9 1 cooling coil function of 2 9 1 description 2 9 12 description of 2 9 12 function of 2 9 1 2 9 12 preparing for installation 3 2 5 screws removing from Pneumatics cabinet 3 2 5 switch system pressure function of 2 9 1 test procedure 3 5 2 valve water drain VL15 function of 2 9 1 configurations 4 PMTs nonswitchable amp See 4 PMTs nonswitchable amp 4 PMTs switchable amp with peak scatter function See 4 PMTs switchable amp with peak scatter function 5 PMTs nonswitchable amp no gated amp See INDEX 5 INDEX 5 PMTs nonswitchable amp no gated amp 5 PMTs switchable amp no gated amp See 5 PMTs switchable amp
53. 2 3 12 block diagrams 3 PMT Sub Amp card 2 11 2 20 40 60 Microsecond Delay card 2 12 6 3 PMT Sub SW R card 2 10 1 4 PMTs Nonswitchable Amps configuration 2 6 10 4 PMTs switchable amp with peak scatter function configuration 2 6 12 5 PMTs Nonswitchable Amps No Gated Amp configuration 2 6 14 5 PMTs switchable amp no gated amp 2 6 16 7 Microsecond Delay card 2 12 6 20 40 60 Microsecond Delay card 2 12 6 ADC and PSH Control card 2 13 6 Bitmap and Sort Decision card 2 8 7 Compressor module 2 9 12 Dual FL Amp card 2 11 3 Dual FL SW R card 2 10 2 Gated Amp Control card 2 12 1 Lister card 2 13 13 Mux and Scope card 2 13 3 Peak ADC PSH Control card 2 13 7 Peak Scatter Mux SW R card 2 10 4 PMT Gated Amp card 2 12 5 Prism and Sort Window Test card 2 8 5 Quad PSH card 2 13 5 sample pressure system new type 2 9 7 sample pressure system old type 2 9 6 Scat Aux Gated Amp card 2 12 3 Scat CV Amp card 2 11 1 Scat CV SW R card 2 10 3 sheath pressure system new type 2 9 11 sheath pressure system old type 2 9 9 Sort Delay card 2 8 9 PN 42370036 INDEX Sort Oscillator card 2 8 10 Sort Output card 2 8 14 Sort subsystem circuit cards functions of 2 8 3 Transistor Bracket card 2 8 14 Video subsystem 2 5 4 blocking filter BK function 2 3 12 BP band pass filter function 2 3 12 bubble drain valve VL7 function of 2 9 2 C cable connections blue ribbon cables 4 10 4 Dual CRT Powe
54. 21 22 23 24 25 26 27 28 29 30 31 32 33 Install miniature fitting actuator pinch valve and hex nut on manifold at right angle to solenoid valve at position 13 Install fittings with nuts on holes marked FF39 FF33 FF42 on mounting plate Install a fitting with a nut on the hole marked FF38 on the mounting plate Route the 7 5 in piece of tubing through the hole nearest the manifold to the new pinch valve at position 13 Connect the tubing to FF38 and FF39 to pinch valve side of the bracket Connect tubing to the fitting at the manifold position 14 Route the other end of the tubing through the hole on the mounting plate towards the backside of the mounting plate Pass tubing through the opening on the top of the pneumatic housing Pass tubing through the frame to be connected later Connect the remaining piece of tubing to FF38 Connect the tubing a Route the other end of tubing through the opening on top of the pneumatic housing b Pass tubing through the frame c Connect fitting to the end of the tubing d Connect 9 inches of the tubing and leave until later Install the I beam tubing through the pinch valve at position 13 to fittings FF33 and FF42 on the pinch valve side of the bracket Remove the waste tubing from FF15 on the rear of solenoid side of the bracket Remove the brass T fitting Connect the tubing from the top of the T fitting coming from FF21 to FF15 Note longer pi
55. 34 kg 75 Ib Due to low output power this laser is not recommended for sense in air operation tAir cooled laser dWater cooled laser Beam Shapers There are 7 beam shaper assemblies available Figure 2 3 2 e 15x60 confocal e 15x80A15 2 3 5 2 INSTRUMENT DESCRIPTION OPTICAL COMPONENTS 15x80 A6 8x80A15 40x80 confocal 10x80A15 2 3 6 Figure 2 3 2 Beam Shaper Options LENGHT FRONT LENS CL LENS HOLDER 7 15 X 60 CONFOCAL 15 80 15 15 80 6 ACTUAL SIZE 15 80 15 40 X 80 CONFOCAL S 10 X 80 15 7003150 Each assembly consists of a sealed pair of crossed cylindrical lenses that produce a specific elliptical beam cross section at the flow cell The lenses are centered in each assembly and allow the user to change beam shapers with only minor adjustments The front lens nearest the flow cell is horizontal and determines the height of the laser beam parallel to the flow The vertical rear lens determines the width of the laser beam orthogonal to the flow Beam height determines peak signal pulse width For doublet discrimination cell size must be at least half the height of the beam Beam width directly affects sensitivity and resolution PN 4237003C INSTRUMENT DESCRIPTION OPTICAL COMPONENTS See Heading 2 7 OPTICAL ELEMENTS for additional information on the optical el
56. 40 microseconds for the 100 p sort sense tip at 12 psi 60 microseconds for the 76 p Sort Sense tip at 12 psi 7 microseconds for the 76 p jet in Air tip PN 4237003 A 1 3 QUICK REFERENCE INFORMATION TOLERANCES AND LIMITS A 1 4 4237003C QUICK REFERENCE A JUMPERS AND SWITCH SETTINGS A 2 JUMPERS AND SWITCH SETTINGS Jumpers for Circuit Cards Used During Installation and Upgrade Procedures 3 PMT Sub SW R2 Card For 5 PMT installation E3 E4 In 1 2 Out E5 E6 Out HV DAC Card Jumper to be HV DAC 2 card for PMT 5 installation E5 E9 E6 E10 E7 Ell E8 E12 HV DAC Control Card E E5 E2 E6 E3 E7 E4 E8 HV DAC Control Card Gated Amp 5 PMT option E5 E9 E6 E10 7 11 E8 E12 Lister AT Card on PC Model 486 E71 E72 Mux and Scope Card For Gated Amp upgrade 1 2 4 Sensor Interface Card For Watchdog timing procedure Remove E25 E26 PN 4237003 2 1 QUICK REFERENCE INFORMATION JUMPERS AND SWITCH SETTINGS Jumpers for Cards in Card Cages Multibus Card Cage Table A 2 1 Multibus Card Cage Jumpers and Switch Settings Card Jumper CPU IBC 86C Intel factory configured Serial 1 0 none Data Taker Interface none 256K Memory card 1 2 3 4 5 6 7 8 SW1 X X X X id the was SW2 0 0 0 0 0 0 0 0 manu actured after 10 30 96 or i sW3 0 0 0 X X 0 X Closed the instrument has the new computer card PN 6706571 see 9
57. Attach the flow cell tip from step 2 to the new flow body c Connect the sheath vacuum and sample tubing to the new flow body Note The kit includes a length of 0 015 in tubing you can use to replace the existing 0 010 in tubing Plug the bimorph connector into the prongs of the Piezo labeled and Verify that the cable protrudes from the instrument Note This is the opposite orientation from how the cable was attached to the old bimorph assembly 10 Do Initial Flow Cell Alignment below 11 Install the beam shaper and do an optical alignment as instructed in the Special Procedures and Troubleshooting manual 12 Do Heading 5 1 VERIFICATION INSPECTION PROCEDURE 3 16 1 INSTALLATION PROCEDURES PIEZOELECTRIC TRANSDUCER INSTALLATION 3 16 2 Initial Flow Cell Alignment 1 2 Remove the beam shaper Verify that the laser beam is centered on the scatter sensor If not or if there is any doubt as to laser alignment install the targets and align the laser Turn Sheath on and open the laser shutter Adjust the flow cell vertically to ensure the laser beam is passing through the quartz cell Rotate the flow cell so the laser beam reflects back through the laser a Loosen the nut that secures the flow cell to the bimorph b Rotate the flow cell as needed c Tighten the nut Adjust the flow cell tilt so the laser beam reflects back through the laser shutter a Loosen the two bolts that secur
58. CH T 901 5106 1 0 73 83 E ey NI Z3 13 Hf 1015106 n 90194 A IN SI NI NI S1VH Du 13 110 NI Lu 106 ir NK NI m vini STI 193 Y MS YMS 4319 Wad Z LINd 0 H9 e INI AH pr 3409 WOI E WALSAS ei 3 DOGO T cuo B A AH gt Q XY09 AH SES XV02 1VNSIS 106 1015 1018 191228 xyeag 9 19 6 972 42370036 2 6 12 INSTRUMENT DESCRIPTION ACQUISITION SUBSYSTEM 5PMTs Nonswitchable Amps No Gated Amp The 5PMT Nonswitchable Amp No Gated Amp Figure 2 6 10 was the first field upgradable configuration With this setup PN 42370036 The system must now subtract from four signals to provide compensation which it does by routing the FL2 and FL3 signals to both PMT Sub SW R cards As a reminder with the 4PMT nonswitchable Amps setup there were only three signals A second 3 PMT Sub SW R card processes the additional PMT The 3 PMT Sub SW R 1 card processes all combinations of FL1 FL2 and FL3 The 3 PMT Sub SW R 2 card processes all combinations of FL2 FL3 and PLA The system cannot compensate for FL1 and FL4 together because they are not routed to a common card This is not a problem however since the two PMTs normally used optical filters with passbands far enough apart to prevent any spectral overlap 2 6 13 2 INSTRUMENT DESC
59. Indicator R indicates not used in Analyzer system Additional Schematics for Analyzer Systems Analyzer Interface Additional Schematics for Autoclone Sorting Option Systems Autoclone Sorting Option card Interconnect Position board Additional Schematics for Gated Amp Systems 7 Microsecond Delay R PMT Gated Amp R Quad 20 40 60 Microsecond Delay R SCAT AUX Gated Amp R Additional Schematics for Non Current Production Systems Pneumatic Interface R before low bleed regulators Scatter Sensor board Sensor Interface R before Panelyzer SCHEMATICS REQUIRED N 6319714 N 6319846 N 6319272 N 6319850 N 6319296 N 6319727 N 6319855 N 6320547 N 6320016 N 6319789 N 6320927 N 6319305 N 6320917 N 6319807 mg mg mg mg mg mg mg wg mg mg mg cm 6321014 6320579 6320653 PN 6320652 PN 6319615 PN 6319342 PN 6319749 PN 6319345 PN 6319758 PN 6319728 PN 6319759 SCHEMATICS l6 SCHEMATICS SCHEMATICS REQUIRED Additional Schematics for Non ESP Systems ADC amp PSH Control Gated Amp Control R2 Sort Delay R Sort Oscillator R PN 6319509 PN 6320143 PN 6319580 PN 6319576 Additional Schematics for Non Switch Amp Jumper Configured Cards Systems 3 PMT Sub Amp R Dual FL Amp R SCAT CV Amp R PN 6320138 PN 6320137 PN 6320136 PN 4237003C CONTENTS ri 7 TROUBLESHOOTING 7 1 1 7 1 ISOLATING THE PROBLEM AREA 7 1 1 Purpose 7 1 1 Tools Supplies Needed 7 1 1 Procedu
60. NO RMALIZATION FRE COM NT M 80 SETTINGS CKPOR CH 2 7003125A 4 2 6 PN 4237003 PN 42370036 SERVICE AND REPAIR PROCEDURES SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE 11 Observe the sort pulses with your scope TP6 on Sort Oscillator card For non ESP systems the front and back porch adjustments are on the Sort Oscillator card a Adjust R93 for the best side streams while keeping the front porch level between 60 and 9596 of the main pulse b Adjust R92 for the best center waste stream while keeping the back porch level between 5 and 3096 of the main pulse Delay Calculation l Once the side streams are stable and singular at the Cytometer Options screen verify that the proper flow cell is selected e 76 e 1004 e jet in air 2 Using the following initial camera setup procedure calculate the droplet delay setting a Panallthe way to the right b Move the beam to the far right c all the way to the left d Ifthe last attached droplet is not visible zoom to the new position and repeat steps a through c e Panright and set the beam and flow cell edge cursors f Pan left and set Last Attached Drop Left Drop and Right Drop cursors g Enter the number of drops between the left and right drop h Record the total equivalent drop value using whole numbers only After calculating the delay in the camera screen go to the Sort screen Run the fluorospheres and set a
61. Optics Alignment Software Improper use Power problem Software bug PN 4237003C TROUBLESHOOTING ISOLATING THE PROBLEM AREA General Troubleshooting Most problems that occur with the system are fluidics related and some of the fluidic problems result in the same symptoms noted in the Optics section of Table 7 1 2 Always check fluidics before attempting an alignment procedure See Heading 7 3 TROUBLESHOOTING FLUIDIC PROBLEMS for details on fluidic troubleshooting Aside from fluidics other troubleshooting may be required See Tables 7 1 2 through 7 1 8 e Table 7 1 2 Autoclone Sorting Option General Troubleshooting Information Table 7 1 3 General Troubleshooting Information e Table 7 1 4 Lasers General Troubleshooting Information e Table 7 1 5 Optics General Troubleshooting Informationx e Table 7 1 6 Electronics General Troubleshooting Information e Table 7 1 7 Software General Troubleshooting Information e Table 7 1 8 Power Supplies General Troubleshooting Information Table 7 1 2 Autoclone Sorting Option General Troubleshooting Information Problem Probable Cause Corrective Action No sheath flow Clogged flow cell 1 Press CLEAR if sheath is fully clogged For partial clogs press SHEATH and VACUUM If still clogged brush tip with a camel hair brush and press SHEATH and VACUUM If still clogged remove flow cell tip and force liquid or air through it
62. Peak GFL e Histogram 7 RFL PMT3 e Histogram 8 PMT4 FALS Discriminator 100 all others OFF Gains 10 HV 400 Set Cytosettings to SEND Ensure all values are transferred then select Acquisition vr Start Ensure that CVs obtained on GFL are comparable to the specifications of the fluorospheres used 5 Ensure via the pulse display that the integral peak and log signals are generated for all PMTs 6 Vary discriminator setting to ensure correct operation Save eight histograms and two Listmode files 8 Use an appropriate protocol and analyze Cyto Trol control cells or samples stained for T4 T8 and B4 Note Be sure to include a PE vs FITC gated on FALS vs 90LS histogram Analysis Check 1 At the Workstation Listmode screen verify operation of the Listmode playback 2 the Workstation Multigraph screen verify operation of stored histogram analysis Sort Check Note This procedure does not apply to Elite Analyzer systems 1 If the instrument has been shut down a Perform the Startup procedure b Check for proper alignment and flow and perform the cleaning and adjustment procedures as needed Clean the deflection plate if saline is evident Plug in the deflection plate 3 6 1 INSTALLATION PROCEDURES SYSTEM TESTING 4 Place the top of the strobe between the bottom of the beam shaping lens and the bottom of the lens holder 5 Center the stream within the sort plates opening by adjusting t
63. SCREW ZOOM SETTING WINDOW SCREW PERISCOPE _ CLAMP Later production cameras use setscrews SECONDARY FOCUS ADJUSTMENT S SSC SCREW BELT POSITION POT N PULLEY POSITION MECHANISM LEFT CRT 7003080A 4237003C SERVICE AND REPAIR PROCEDURES SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE 4 2 SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE Purpose This procedure describes how to verify and adjust the Crystal Drive and Sort Pulse signals Perform this procedure when any of the following conditions occur e System installation Replacement of Sort Oscillator card Sort Output card 90 V power supply 36 V power supply or sort transistor Sort problems See Figure 4 2 1 Figure 4 2 1 Waveforms SORT PULSE WAVEFORMS Center Level Front Porch Back Porch Center Level Three drops Amplitude Center level 2 x 31 25 62 5 Microseconds 0 Volts Center Level Front Porch Back Porch Two drops Center level 31 25 Microseconds Front Porch One drops Back Porch Center level CRYSTAL DRIVE WAVEFORM Peak to Peak Amplitude gt Cycle One droplet 31 25 Microseconds 7003166A Regarding Figure 4 2 1 Set Drive Frequency to 32 kHz At this frequency each droplet period will be 31 25 microseconds Set the Crystal Drive to 100 Set the Deflection amplitude to 100 Set Sort Test Sort Right Enable Sort Left Ena
64. VL3 function of 2 9 2 sample pressure troubleshooting problems 7 4 3 sample pressure system new type block diagram 2 9 7 description 2 9 7 sample pressure system old type block diagram 2 9 6 DAC reference voltage measurement of 2 9 4 description 2 9 4 1 function of 2 9 4 TP8 function of 2 9 4 sample pressure valve VL4 function of 2 9 2 sample regulator adjustment See pneumatics sample requirements 2 1 2 satellite droplets illustration 7 5 8 Scat CV Amp card signal flow summary of 2 6 9 Scat Aux Gated Amp card block diagram 2 12 3 function 2 12 2 inputs 2 12 2 jumpers 4 10 3 A 2 3 Scat Aux Gated Amp R card jumpers 3 9 4 Scat CV Amp card block diagram 2 11 1 function 2 11 1 Scat CV Amp SW R card jumpers 4 10 2 A 2 2 Scat CV SW R card block diagram 2 10 3 function 2 10 2 jumpers 3 9 4 Scat CV SW RI card jumpers 4 10 3 A 2 3 14 INDEX Scatter Sensor R Pre Amp card replacing with Peak Scatter Sensor card for Gated Amp installation schematics engineering included in this manual 6 1 1 sensitivity test procedure 3 15 2 sensor area components 2 1 2 Sensor I O card function 2 5 8 Sensor Interface card C44 location of 3 17 1 communication pathways from Acquisition card cage 2 9 4 jumpers 4 10 2 4 10 3 A 2 1 A 2 2 A 2 3 jumpers for Watchdog timing 3 17 1 modifying for Watchdog timing 3 17 1 R77 location of 3 1 7 1 sensors cells through laser measuring 2 13 1 optic
65. and to provide precise temperature compensation Figure 2 6 4 Circuit Operation R1 P En A S I ade Vv 7003034A Integration The Elite uses a standard operational amplifier integrator circuit to measure the sum of all the fluorescence in the cells The circuit uses a capacitor in the op amp feedback to sum the input voltage over time This results is an output voltage proportional to the area under the input pulse The component values of the circuit determine the length of time that the integrator can charge 20 microseconds for quartz flow cells 5 microseconds for jet in air flow cells PN 4237003C PN 42370036 INSTRUMENT DESCRIPTION ACQUISITION SUBSYSTEM Once the input pulse passes the integrator discharges The discharge provides the characteristic tail to the integral signals See Figure 2 6 5 Figure 2 6 5 Integrator Discharge Peak Signal Integral Signal TIME INTEGRATION CHARGE TIME DISCHARGE TIME Volts Out the area under Volts in our 7003031A Special Functions Switchable Amplifiers Bandwidth The component values of the integrator circuits must match the expected input pulse width Depending upon the flow cell tip the pulse width varies The pulse width is 20 microseconds for the sort sense flow cell 76 or 100 tip e 5 microseconds for the jet in air flow cell tip The integrators have two sets o
66. range of 2 7 1 LINE 4 3 1 line voltage jumpers A 2 4 location 4 3 1 selection 4 3 1 line voltage jumpers See jumpers list of parts See parts list Lister A B card function 2 13 14 Lister AT card jumpers 4 10 4 A 2 1 Lister card block diagram 2 13 13 Lister Out card block diagram 2 13 13 control circuits function 2 13 13 FIFO register function 2 13 14 10 INDEX function 2 13 12 input buffer function 2 13 14 output buffer function 2 13 14 time circuits function 2 13 13 log amplification clamping diode function 2 6 3 description 2 6 2 equation 2 6 3 illustration log amp to input voltage 2 6 3 input voltage 2 6 3 output voltage 2 6 3 response in log lin axes 2 6 3 log amplification equation 2 6 3 long pass LP filter function 2 3 12 LP long pass filter function 2 3 12 Lx marker See Autoclone Sorting Option 2 2 3 Ly See Autoclone Sorting Option 2 2 3 manual field service conventions used 1 1 3 format 1 1 2 intended audience 1 1 1 layout 1 1 1 organization description 1 1 2 page numbers format of 1 1 2 safety warnings description of 1 2 1 scope of 1 1 1 special headings defined 1 1 3 markers CC defined 2 2 3 CW defined 2 2 3 Hx marker defined 2 2 3 Hy marker defined 2 2 3 Lx marker defined 2 2 3 Ly defined 2 2 3 Motor Control card function 2 5 8 potentiometer speed control wiring of 2 5 8 Multibus CPU system damage preventing 2
67. 00 psi on the screen Adjust R34 for exactly 10 00 Vdc at TP8 Remove the aluminum seal from regulator and adjust the SPAN potentiometer for exactly 15 00 psi on your gauge Set the Sample Pressure to 3 00 psi Verify that the DVM reads 2 00 0 04 Vdc If the DVM reads differently replace the Pneumatic Interface card With TP8 reading 2 00 0 04 Vdc adjust the ZERO potentiometer on the regulator until the gauge reads 3 00 psi Repeat steps 3 through 8 until no further adjustments are needed Disconnect the DVM from TP8 Replace the seal on the regulator Sheath Regulator QM ME ob E N 10 11 12 13 Connect the DVM to TP11 Remove the jumper from El to E2 and place E2 to Connect the external gauge to the sheath pressure line Set the Sheath Pressure to 15 00 psi on Cytometer Control screen Adjust R81 for exactly 10 00 Vdc at TP11 Remove the aluminum seal from regulator and adjust the SPAN potentiometer for exactly 15 00 psi on your gauge Set the Sheath Pressure to 3 00 psi Verify that the DVM reads 2 00 0 04 Vdc If the DVM reads differently replace the Pneumatic Interface card With TP11 reading 2 00 0 04 Vdc adjust the ZERO potentiometer on the regulator until the gauge reads 3 00 psi Repeat steps 4 through 9 until no further adjustments are needed Remove the DVM from 11 Replace the seal on the regulator Do Pneumatic Interface Card Adjustment 4 5 3 4 SERVIC
68. 1 card and 2 card for Software Version 3 1 and below Jumper E3 E4 In e Jumper El E2 E5 E6 Out Install 3 Sub Amp Amp SW R2 card in the slot to the left of the existing 3 Sub Amp Amp 1 card Quad 20 40 60 Microsecond Delay card slot Cabling Configuration and Installation Remove the following bullet connectors from 3 PMT Sub Amp SW R card 1 Int Out J8 Relabel as 3 Int FL3 Peak Out J6 Relabel as 3 Pk FL3 Log Out J7 Relabel as 3 Log Remove the following bullet connectors from the Dual FL SW R card e FI2 Int Out J7 Relabel as PMT 4 Int e FI2 Peak Out J6 Relabel as 4 Pk FL2 Log Out J8 Relabel as 4 Log Remove bullet connectors from 3 PMT Sub Amp SW R 1 FL2 out J5 Relabel cable ends as PMT 4 Out and PMT 4 In and connect the cables a PMT 4 Out end to Sub Amp SW R 2 FL2 out J5 b PMT 4 Inend to Dual FL SW R 2 FL2 In J5 Connect a BNC T connector to PMT 3 Signal coax Connect a BNC T connector to PMT 4 Signal coax Connect two short BNC bullet coax cables to PMT 3 BNC T connector and label bullet ends as PMT 3 PN 4237003C 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 INSTALLATION PROCEDURES 5TH PMT KIT WITH SWITCHABLE AMPS INSTALLATION PROCEDURE Remove signal coax from 3 PMT Sub Amp SW R 1 FL3 in J3 Relabel as 5 and connect e One PMT 3 signal coax to Sub Amp SW R
69. 10 6 4 10 7 CPU Card Jumper Locations 4 13 1 Pulse Generator Card Values 4 2 15 Line Voltage Jumpers 4 3 1 Power Supply Measurement and Adjustments 4 4 2 Potentiometer Adjustments 4 5 1 Back Panel Wiring Configuration 4 10 1 Multibus Card Cage Cards Jumper and Switch Settings 4 10 1 Non Gated Amp Card Cage Jumper Setting 4 10 2 Gated Amp Card Cage Jumper Settings 4 10 3 Data Acquisition Card Cage Cards Jumper Settings 4 10 3 Cable Connections 4 10 4 Cable Connectors 4 10 10 PN 4237003C SERVICE AND REPAIR PROCEDURES 4 1 OPTICAL ALIGNMENT PROCEDURE System Alignment Purpose The alignment procedures depend on the types of lasers installed and their optical configuration This procedure applies to the standard configuration which includes the air cooled Argon and HeNe co linear beam configuration Refer to the appropriate procedure for other laser configurations e EPICS Elite Options Manual gt HeCd Model 56 alignment gt HeCd Model 74 alignment gt Innova 305 alignment gt Innova 90 5 alignment e Heading 3 7 OPTIONAL LASER INSTALLATION contains procedures for aligning HeCd Innova 305 Innova 90 and HeCd lasers e Heading 3 14 ENTERPRISE LASER UPGRADE contains procedures for Enterprise laser alignment Heading 3 9 GATED AMPLIFIER UPGRADE contains procedures for alignment for Gated Amp operation WARNING Risk of personal injury due to exposure to Laser beam Do not proceed until y
70. 100 Set Drops Sorted to 3 Adjust Stream offset so the signal baseline is 0 Vdc Connect the oscilloscope to the Sort Output card TP2 Adjust R94 on the Sort Oscillator card for a baseline to center level value of 4 0 0 1 V Connect the oscilloscope to the flow cell sample insertion rod Adjust R119 on the Sort Output card for a baseline to center level value of 80 1 V Phase Settings 1 2 3 Connect one scope channel to the crystal drive and the other to the sort pulses Trigger on the sort pulse channel Vary the phase and verify a corresponding shift in the crystal drive with respect to the sort pulses Note Each phase increment 0 0 to 0 9 equals a one tenth wavelength shift of the crystal drive sinewave signal Delay Settings 1 2 PN 42370036 Acquire beads and set up the windows necessary to obtain sort pulses Turn Sort Test OFE 4 2 3 4 SERVICE AND REPAIR PROCEDURES SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE 3 Sortall the beads to the right that are in the GFL histogram a Select GFL for the lower Digiscope trace b Leave one scope channel connected to the insertion rod and connect the other channel to the GFL test point on the lower trace LT BNC connector on the Aux power panel c Besure you can see both pulses on the scope then set the trigger to Sync on the GFL pulse The sort pulse displayed on the scope is delayed by the Delay entered on the touch screen 4 Ver
71. 13 J6 J9 Dual FL SW R slot 3 J6 Scat CV SW R1 slot 14 J6 J8 PMT Gated Amp slot 6 Scat Aux Gated Amp slot 11 J2 J4 J3 J5 Peak Scatter Mux SW slot 13 Scat CV SW R1 slot 14 J12 J10 Scat Aux Gated Amp slot 11 Gated Amp Control R3 slot 1 J3 PMT Gated Amp slot 6 Gated Amp Control R3 slot 1 J4 Bundle these cords and continue PN 42370036 PN 42370036 Table 3 9 2 Coax Cable Interconnections INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE From To Mux and Scope PSH 1 slot 7 2 J3 J3 J5 J4 J7 PSH 1 slot 7 PSH 2 slot 8 J5 J6 J3 J7 J5 J8 J7 Mux and Scope Lower Trace LT J29 Mux and Scope Upper Trace UT 30 Mux and Scope No Connection J26 Mux and Scope Scat CV SW R 1 slot 14 J10 J4 Mux and Scope Scat CV SW R 1 slot 14 J11 J3 J12 J2 Mux and Scope 3PMT Sub SW R 1 slot 5 J13 J8 J14 J7 Mux and Scope Peak Scatter Mux SW slot 13 J15 J4 J9 J14 Mux and Scope Dual FL SW R slot 3 J16 J3 J17 J4 Mux and Scope Dual FL SW R slot 3 J18 J2 J19 J7 J20 J8 Mux and Scope Scat CV SW R1 slot 14 J21 J9 Mux and Scope 3PMT Sub SW R2 slot 4 J22 J8 J23 J7 Mux and Scope Peak Scatter Mux SW R slot 13 J24 J10 Bundle these cords and continue 3 9 7 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE Table 3 9 2 Coax Cable Interconnections
72. 18 Table 8 2 18 Analyzer Top Inside View Detail of Parts See Figure 8 2 18 Item Part Number Description Item Part Number Description 1 6803642 Double acting pinch valve 13 6216012 Spacer pneumatic 0 562 od x 0 500 i d 2 2822033 Nut 47 32 UNS x 0 562 AFX 14 6211015 Pilot actuator 3 1018879 Plunger test tube 15 6028086 Fluid detector cable 4 2515062 Spring 0 344 o d x 0 62 x 16 2839043 Self locking screw 6 32 x 0 62 0 25 5 6858369 Sensor card mount 17 6232086 Union hose barb 0 062 i d 6 6703953 Pump motor detector 18 2523062 O ring seal 0 187 i d 7 6232205 Fitting hose barb elbow 19 2821010 Self locking nut 6 32 x 0 25 AFX 0 125 i d to 10 32 nylon white 8 2523062 O ring seal 0 187 i d 20 2839039 Self locking screw 6 32 x 0 37 9 6232124 T fitting miniature 10 32 21 1020901 Pinch valve bracket 10 6216345 Gasket 10 black 22 6858370 Pinch valve mount bracket 11 1017501 Pinch valve mount 23 6211015 Pilot actuator 12 6855763 Standard pinch valve pull apart 8 2 18 PN 4237003C PARTS LISTS 8 ILLUSTRATED PARTS Figure 8 2 19 Flow Cell Area Elite See Table 8 2 19 2 3 7003101 Table 8 2 19 Elite Flow Cell Area Detail of Parts See Figure 8 2 19 Item Part Number Description 1 6857388 Double acting pinch valve 6211015 Pilot actuator 2 3814172 Flow cell stage 3 3814173 Beam stage 4 6857133 Shutter assembly 5 6856696 Beam
73. 2 2 6 2 Log Amp Response 2 6 3 2 6 3 Log Amp Response Log Lin Axes 2 6 3 2 6 4 Circuit Operation 2 6 4 2 6 5 Integrator Discharge 2 6 5 2 6 6 Typical Switch Amp Integrator Circuit 2 6 6 2 6 7 Gated Amp Concept 2 6 7 2 6 8 4PMT Non Switchable Amps without Peak Scatter 2 6 10 2 6 9 4PMT Switchable Amps with Peak Scatter 2 6 12 2 6 10 5PMT Non Switchable Amps without Peak Scatter 2 6 14 2 6 11 5PMT Switchable Amps with Peak Scatter 2 6 16 2 6 12 Block Diagram Gated Amplifier 2 6 19 2 7 1 Refraction of Light 2 7 1 2 7 2 Positive Lens Focus 2 7 2 2 7 3 Image Information 2 7 3 2 7 4 Negative Lens Focus 2 7 3 2 7 5 Optical Accessories 2 7 5 2 7 6 Beam Shaping Optics 2 7 6 2 7 7 Beam Expander Reducer 2 7 7 2 7 8 Laser Beam Refraction 2 7 8 xviii 4237003 PN 42370036 2 8 1 2 8 2 2 8 3 2 8 4 2 8 5 2 8 6 2 8 7 2 8 8 2 8 9 2 8 10 2 8 11 2 8 12 2 9 1 2 9 2 2 9 3 2 9 4 2 9 5 2 9 6 2 10 1 2 10 2 2 10 3 2 10 4 2 11 1 2 11 2 2 113 2 12 1 2 12 2 2 123 2 12 4 2 12 5 2 13 1 2413 2 2 133 2 13 4 2 13 5 2 13 6 2 1357 2 13 8 2 13 9 3 1 1 3 2 1 3 2 2 3 2 3 3 2 4 3 2 5 3 2 6 3 2 7 3 3 1 3 4 1 3 4 2 CONTENTS Sort Subsystem Overview 2 8 2 Block Diagram Sort Subsystem Card Functions 2 8 3 Block Diagram Prism and Sort Window Test Card 2 8 5 Block Diagram Bitmap and Sort Decision Card 2 8 7 Block Diagram Sort Delay Card 2 8 9 Block Diagram Sort Oscillator Car
74. 2 2 2 3 switch overtravel function of 2 2 2 testing the full operation 3 10 9 testing the mechanism 3 10 7 testing the stepping pattern 3 10 8 tray position markers defined for 2 2 3 tray rotation arm movement 2 2 2 troubleshooting problems with 7 1 3 valve waste VL14 function of 2 9 2 version 3 25 software serial numbers of instruments with 3 18 1 waste catcher adjustments 3 10 9 Autoclone Sorting Option card function 2 2 3 limit switches 2 2 3 Autoclone Sorting Option Interconnect card function 2 2 3 back porch 7 5 7 band pass BP filter function 2 3 12 barrier protection advising the customer to use 1 2 9 batteries Lithium on 256K Memory card 2 5 2 beam expander reducer 2 3 8 function 2 7 7 beam shapers function 2 3 6 2 7 5 sizes available 2 3 6 spot dimensions 2 3 7 typical applications of 2 7 6 beam steering optics function 2 3 7 illustration alignment mirror 2 3 8 beam translator alignment procedure 4 1 3 description 2 3 8 upgrade requirements 3 12 1 PN 4237003C bimorph crystal drive setting 3 15 2 result if unstable 7 5 7 See also Piezo bimorph See Piezo bimorph biofilter clogs affect of 2 9 1 drip chamber function in 2 9 1 Bitmap and Sort Decision card block diagram 2 8 7 digital values reading 2 13 1 function 2 8 5 jumpers 4 10 3 A 2 3 sort left or right determining 2 8 8 bitmap defined 2 8 6 BK blocking filter function
75. 2 36 10 6857286 Cover laser interlock assembly 6857334 Interlock laser with red pin 6857347 Duct intake laser 5 2 35 15 6857424 Duct plenum HeNe laser 8 2 35 1 6857483 Interlock defeat bar red L shaped 6857515 HeNe laser 8 2 35 4 6857516 Argon laser tested 8 2 35 13 6857611 Stage beam splitter 488 nm 8 2 35 10 6857612 Mirror HeNe laser 8 2 35 11 6857613 Stage beam splitter 325 nm 6858243 Beam splitter 633 nm light HeNe laser 6858244 Beam translator UV light either HeCd or Argon UV laser 6858245 Beam translator 488 nm light Argon laser 6858459 Target multipositional 3 hole large 8 2 36 11 6912654 Kit Laser HeCd 74 Omnichrome 6912749 Laser red HeNe Uniphase 8 2 36 1 6912750 Laser air cooled Argon Cyonics 8 2 36 2 6912792 Laser Innova 305 Coherent 8 2 36 3 6912834 Kit HeCd 74 laser 8 2 38 1 6913199 Laser Enterprise 6913211 Laser Spectrum 6913212 Laser He Ne green Melles Griot 6913229 Laser Coherent 6913248 Heat exchanger Enterprise 230 V 6913249 Heat exchanger Enterprise 115 V 7232344 Diode Enterprise laser starter 7232500 Enterprise laser laser head assembly 71232501 Enterprise laser power supply assembly 7232502 Enterprise laser heat exchanger 115 Vac 7232503 Enterprise laser remote control 7232504 Enterprise laser remote defeat 1232505 Enterprise laser heat exchanger defeat PN 4237003C 8 1 17 8 PARTS LISTS MASTER PARTS LIST IN NUMERICAL ORDE
76. 2 5 4 SERVICE AND REPAIR PROCEDURES SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE 9 Check the side streams with a sample running Turn off Sort Test Load your sample protocol Set Sort Setting to Receive Set Cytometer Settings to Send IMPORTANT Risk of compromising sort stability sample flow if sample flow is adjusted more than 10 units above the optimal setting for running beads Avoid compromising sort stability by not adjusting the sample flow more than 10 units above the optimal setting for running beads e Change the Sort Settings to Send to send your sample sort gates Run the actual sample at the data rate desired for sorting Note the differential pressure required to achieve the desired sort rate and use this same Diff pressure when doing the sort matrix checking the delay setting OR Run fluorospheres at the same concentration as the sample to be sorted 10 Change the Sort Droplet to 5 and ensure that the side and center streams are as clean as possible by adjusting the front and back porch adjustments Figure 4 2 2 Note change in sheath pressure may require you to readjust the porches even if no other changes were made e Check the side and center streams on the 1 2 and 3 Drops Sorted settings The streams should look good on all of these settings the side streams do not look good clean or change the flow cell Figure 4 2 2 Front and Back Porch BEAM wip
77. 2 9 6 8 2 11 8 6703941 DAC card 6704008 Gated Amp 8 2 8 6 8 2 9 33 6704009 Scat Aux Gated Amp 8 2 8 11 8 2 9 28 6704089 Dual CRT Controller 8 2 9 8 2 11 6704139 ADC and PSH Control Card 8 2 12 6704142 Data Lister Out 8 2 8 23 8 2 9 15 8 2 12 10 6704147 Interface and Scaler 8 2 8 20 8 2 9 17 8 2 12 13 6704164 Mux and Scope Interface 8 2 8 27 8 2 9 11 23 8 2 12 6 6704169 Quad Peak Sense and Hold PSH 1 8 2 8 26 8 2 9 13 8 2 12 8 PN 4237003C MASTER PARTS LIST IN NUMERICAL ORDER CATEGORY Table 8 1 4 Cards Continued PARTS LISTS Part Number Description Figure Item 6704169 Quad Peak Sense and Hold PSH 72 8 2 8 25 8 2 9 12 8 2 12 7 6704185 Prism and Sort Window 8 2 8 22 8 2 9 16 8 2 12 11 6704186 Bitmap and Sort Decision 8 2 8 21 8 2 12 12 6704188 Sort Oscillator 8 2 9 20 8 2 12 15 6704198 Sort Delay 8 2 9 19 8 2 12 14 6704201 Sort Output 8 2 8 17 8 2 9 21 8 2 12 16 6704241 Quad 7 Microsecond Delay 71 8 2 8 9 6704241 Quad 7 Microsecond Delay 2 8 2 8 8 6704327 36 V Regulator 8 2 13 1 6704441 KIR 3 8 2 7 2 6704463 Dual Laser Control 8 2 9 7 8 2 11 9 6704552 Pneumatic Interface Card Used only with Norgren pressure regulators 8 2 4 3 6704660 Camera Interface 8 2 9 8 8 2 11 6 6704665 Lister AT A amp B 6704679 Motor Controller 8 2 13 5 6704697 PMT Gated Amp R 6704798 Pin Hole Driver 6704893 Gated Amp Control 8 2 8 2 6704904 Quad 20 40 60 Microsecond Del
78. 3 1 3 7 5 3 test points Pneumatic Interface card 4 5 1 1 Pneumatic Interface card 4 5 1 A 1 2 TP10 Pneumatic Interface card 4 5 1 A 1 2 TP2 Pneumatic Interface card 4 5 1 A 1 2 TP3 Pneumatic Interface card 4 5 1 A 1 2 TP5 Pneumatic Interface card 4 5 1 A 1 2 8 sample pressure system old type 2 9 4 test procedures sensitivity 3 15 2 test procedures performance of 5 PMT operation 3 11 4 analysis 3 6 1 camera 3 5 3 Compressor module 3 5 2 computer 3 5 4 Cytometer Control Computer 3 5 1 lasers 3 5 4 pneumatics 3 5 2 system 3 6 1 theories of operation wave propagation 2 7 5 Thin Lens equation 2 7 4 time optimum PN 4237003C setting 2 13 8 timing ASD illustration of 2 13 12 differences in 2 13 1 Watchdog procedure 3 17 1 TOF PPU card verifying sort coincidence after replacing the card 4 2 9 TP See test points training See customer training transformers laser auto 100 Vac input to 4 3 1 laser auto 115 Vac input to 4 3 1 laser auto 230 Vac input to 4 3 2 laser auto 240 Vac input to 4 3 2 main system 100 Vac input to 4 3 1 main system 115 Vac input to 4 3 1 main system 230 Vac input to 4 3 1 main system 240 Vac input to 4 3 1 Transistor Bracket card block diagram 2 8 14 function 2 8 19 trigger level defined 2 12 2 troubleshooting Acquisition subsystem 7 2 1 Autoclone Sorting Option 7 1 3 blockages 7 3 1 color compensation 7 2 3 electronics 7 1 6 general
79. 3 9 16 AUX 2 Adjustment 3 9 16 PMT 2 Adjustment 3 9 17 PMT 3 Adjustment 3 9 17 PMT 4 and 5 Adjustment 3 9 18 Laser Alignment With Gated Amplifier 3 9 22 Laser Configuration 3 9 22 Laser Alignment 3 9 23 Calibration 3 9 24 AUTOCLONE SORTING OPTION INSTALLATION 3 10 1 Purpose 3 10 1 Tools Supplies Needed 3 10 1 Electrical and Mechanical Inspection 3 10 2 Preliminary Procedure 3 10 2 Upgrading the Instrument to be Autoclone Sorting Option Ready 3 10 3 Calibration Procedure 3 10 6 Adjusting Autoclone Sorting Option Speed 3 10 7 PN 4237003C PN 42370036 3 12 3 13 AH 219 3 16 CONTENTS Testing the Autoclone Sorting Option Mechanism 3 10 7 Prepare the System 3 10 7 First Test 3 10 8 Second Test 3 10 9 Stream and Waste Catcher Adjustment 3 10 9 Operational Test 3 10 9 5TH PMT KIT WITH SWITCHABLE AMPS INSTALLATION PROCEDURE 3 11 1 Purpose 3 11 1 Tools Supplies Needed 3 11 1 Hardware Configuration and Installation 3 11 1 Cabling Configuration and Installation 3 11 2 Software Installation 3 11 4 System Test 3 11 4 BEAM TRANSLATOR UPGRADE 3 12 1 Tools Supplies Needed 3 12 1 Laser Mounting Procedure 3 12 1 Standard Laser Alignment Procedure 3 12 1 Collinear Laser Alignment Fine Tuning Procedure 3 12 4 Time Space Separated Laser Alignment Procedure 3 12 5 SORT PERFORMANCE ESP UPGRADE 3 13 1 Purpose 3 13 1 Tools Supplies Needed 3 13 1 Preliminary Procedure 3 13
80. 3 9 25 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE 3 9 26 PN 4237003 INSTALLATION PROCEDURES 3 AUTOCLONE SORTING OPTION INSTALLATION 3 10 AUTOCLONE SORTING OPTION INSTALLATION Purpose Use this procedure to install the Autoclone Sorting Option mechanism Figure 3 10 1 Figure 3 10 1 Autoclone Sorting Option Mechanism 7003149A Tools Supplies Needed Autoclone Sorting Option Kit PN 6912869 6 Connector mounting block PN 1016713 2 Cable assembly PN 6028268 4 Phillips head screw PN 2804039 3 Foam for panel PN PN 3203037 8 Solenoid valves PN 6232096 6 Fitting PN 6232086 9 with seal PN 2523062 1 for manifold Miniature fitting PN 6232413 9 for solenoid valve position 13 Actuator PN 6211015 5 for solenoid valve position 13 Pinch valve PN 6855763 1 for solenoid valve position 13 Hex nut PN 2822033 2 for solenoid valve position 13 Fittings PN 6232104 1 with nuts PN 2822072 3 for holes FF39 FF33 and FF42 on mounting plate Fitting PN 6232417 1 with nut PN 2822016 2 for hold FF38 on mounting plate 7 5 in 19 cm tubing PN 3203015 7 Tubing PN PN 3213063 1 for manifold position 14 Fitting PN 6232109 1 for end of tubing Tubing 9 in 23 cm PN 6303015 5 I beam tubing PN 3213136 1 OUOOOOOODOOCOD Oooocoo PN 4237003C 3 10 1 INSTALLATION PROCEDURES AUTOCLONE SORTING OPTION INSTALLATION 3 10 2 Tubing longer piece 3202039
81. 31 8 2 31 8 2 32 Elite Flow Cell Stage Detail of Parts See Figure 8 2 32 8 2 32 8 2 33 Model 74 HeCd Laser Mounting Hardware Detail of Parts See Figure 8 2 33 8 2 33 8 2 34 Argon Laser Mounting Hardware Detail of Parts See Figure 8 2 34 8 2 34 8 2 35 HeNe Laser Mounting Hardware Detail of Parts See Figure 8 2 35 8 2 35 8 2 36 Laser Placement Top View Detail of Parts See Figure 8 2 36 8 2 36 8 2 37 Cytometer Inside Rear View Detail of Parts See Figure 8 2 37 8 2 37 8 2 38 Omnichrome HeCd Laser See Figure 8 2 38 8 2 38 PN 4237003 8 1 8 iv PN 4237003 PARTS LISTS ra 8 1 MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY The parts are listed alphabetically by category and in order by part number See Table 8 1 1 through 8 1 23 Table 8 1 1 Autoclone Sorting Option Part Number Description Figure Item 1007908 Pin threaded 8 2 24 6 1018230 Bellows 8 2 24 5 1018721 Cover tray 8 2 23 11 1020199 Tray 8 2 23 12 1022047 Holder cylinder 8 2 24 1 1022048 Shaft Autoclone Sorting Option beak 8 2 24 2 1022049 Plate Autoclone Sorting Option beak 8 2 24 3 2508022 Coupling shaft 8 2 23 5 3501018 Motor stepper 1 8 degree angle 0 44 shaft length 8 2 23 4 3505035 Motor stepper 1 8 degree angle 5 5 Vdc 8 2 23 13 4703037 Resistor 4 99 kOhm 4703052 Resistor 10 kOhm 4818006 Switch opto 0 200 gap 8 2 23 9 4836701 Switch opto 0 1 gap 8 2 23 7 5103013 Switch snap action 8 2 23 6 541537
82. 4 2 6 5 Integrator Discharge 2 6 5 2 6 6 Typical Switch Amp Integrator Circuit 2 6 6 2 6 7 Gated Amp Concept 2 6 7 2 6 8 4PMT Non Switchable Amps without Peak Scatter 2 6 10 2 6 9 4PMT Switchable Amps with Peak Scatter 2 6 12 2 6 10 5PMT Non Switchable Amps without Peak Scatter 2 6 14 2 6 11 5PMT Switchable Amps with Peak Scatter 2 6 16 2 6 12 Block Diagram Gated Amplifier 2 6 19 2 7 1 Refraction of Light 2 7 1 2 7 2 Positive Lens Focus 2 7 2 2 7 3 Image Information 2 7 3 2 7 4 Negative Lens Focus 2 7 3 2 7 5 Optical Accessories 2 7 5 2 7 6 Beam Shaping Optics 2 7 6 2 7 7 Beam Expander Reducer 2 7 7 2 7 8 Laser Beam Refraction 2 7 8 2 8 1 Sort Subsystem Overview 2 8 2 2 8 2 Block Diagram Sort Subsystem Card Functions 2 8 3 2 8 3 Block Diagram Prism and Sort Window Test Card 2 8 5 2 8 4 Block Diagram Bitmap and Sort Decision Card 2 8 7 2 8 5 Block Diagram Sort Delay Card 2 8 9 2 8 6 Block Diagram Sort Oscillator Card 2 8 10 2 8 7 Sort Envelope 2 8 11 2 8 8 Block Diagram Sort Output Card and Transistor Bracket Card 2 8 14 2 8 9 Interconnection of Pulse Pileup Det TOF Card 2 8 15 2 8 10 TOF Function of Pulse Pileup Det TOF Card 2 8 16 2 8 11 PPU Function of Pulse Pileup Det TOF Card 2 8 18 2 8 12 Power Supplies Location 2 8 20 2 9 1 Valve Operation 2 9 3 2 9 2 Block Diagram Sample Pressure System Old Type 2 9 6 2 9 3 Block Diagram Sample Pressure System New Type 2 9 7 2 9 4 B
83. 4 5 4 Purpose 4 5 4 Tools Supplies Needed 4 5 4 Procedure 4 5 4 ELECTRONIC PRESSURE REGULATOR ADJUSTMENT 4 6 1 Purpose 4 6 1 Tools Supplies Needed 4 6 1 Procedure 4 6 1 PRESSURE SWITCH ADJUSTMENT 4 7 1 Purpose 4 7 1 Tools Supplies Needed 4 7 1 Procedure 4 7 1 WORKSTATION AND SOFTWARE 4 8 1 PC Model 486 Setup Procedure 4 8 1 SOFTWARE INSTALLATION 4 9 1 Purpose 4 9 1 Tools Supplies Needed 4 9 1 Procedure 4 9 1 ELECTRONIC ADJUSTMENTS CABLE AND JUMPER CONFIGURATIONS 4 10 1 Purpose 4 10 1 Tools Supplies Needed 4 10 1 Cable Configurations 4 10 1 Back Panel Wiring 4 10 1 Jumper Configurations 4 10 1 Multibus Card Cage 4 10 1 Non Gated Amp Card Cage 4 10 2 Gated Amp Card Cage 4 10 2 Data Acquisition Card Cage Configuration 4 10 3 Analyzer HV DAC Control Card 4 10 4 PC Model 486 4 10 4 PN 4237003C 4 Cable Connections 4 10 4 Procedure 4 10 4 4 11 SORT SENSE FLOW CELL ADJUSTMENT AND VERIFICATION 4 11 1 Purpose 4 11 1 Tools Supplies Needed 4 11 1 Procedure 4 11 1 Replacing Metal Fittings with PVC Fittings 4 11 2 Flow Cell Replacement and Verification 4 11 4 4 12 OPTICAL ALIGNMENT PROCEDURE 4 12 1 Purpose 4 12 1 Tools Supplies Needed 4 12 1 Flow Cell Alignment 4 12 1 Parabolic Mirror on Tip Optional 4 12 4 Laser Beam Alignment 4 12 5 Optical Fine Tuning 4 12 8 PMT4 System 4 12 9 5 System 4 12 11 4 13 EXTERNAL MEMORY CARD REPLACEMENT 4 1
84. 4 6 ELECTRONIC PRESSURE REGULATOR ADJUSTMENT and Heading 4 7 PRESSURE SWITCH ADJUSTMENT PN 4237003 4 5 5 SERVICE AND REPAIR PROCEDURES PNEUMATICS SYSTEM 4 5 6 PN 4237003 4 6 SERVICE AND REPAIR PROCEDURES ELECTRONIC PRESSURE REGULATOR ADJUSTMENT ELECTRONIC PRESSURE REGULATOR ADJUSTMENT Purpose This procedure applies only to non low bleed old type regulators Although electronic regulators are factory set field adjustments may occasionally be required to set the Range and Zero adjustments on the electronic regulator as follows 1Vinat the regulator produces 3 psi out 10 V in at the regulator produces 15 psi out Tools Supplies Needed DVM Procedure 1 Connect the DVM across R63 the Pneumatic Interface card 2 Connect external pressure gauge with a Y connector to the sample pressure pinch valve at the far left fitting on the valve bracket 3 At the Valves screen a Adjust Sample SET pressure until the external gauge reads 15 psi b Adjust the Range control until the DVM reads 10 0 V Note The maximum pressure the Pneumatic Interface card will accept is 15 psi If the external gauge reads less than 15 psi when 15 psi is entered in this step adjust R81 to obtain 15 psi c Adjust Sample SET pressure until the external gauge reads 3 psi d Adjust the Zero control until DVM reads 1 0 V Repeat step 3 until no further adjustment is needed Connect the DVM across R22 on the Pneumatic I
85. 4 Block Diagram Peak ADC PSH Control Card 45V DISCSAT INSTRUMENT DESCRIPTION CONTROL DATA FROM CPU ratio inputs Discsat Extend of parameters CONTROL LOGIC These signals control Quad PSH Card operation OPEN CLOSE EM Control signals ay gt PARAMETER ID QUAD REFERENCE i Select parameters for Ratio Sequence parameters lt VF Cos DISCRIMINATOR through A to D circuit NNNM gt DATA READY DACs DATAPATH BUS ux Y v v ON BACKPLANE ANALOG TO Selects one DIGITAL mmm Data Latch output from CONVERTER 16 inputs J2 Inputs from J3 Quad 5 f cards 6 MUX NUMERATOR J7 Selects one d 48 output from J9 16 inputs RATIO ANALOG CALCULATION CIRCUIT J12 MUX J13 J14 Selects one DENOMINATOR up output from J16 16 inputs S J17 7003073A As shown in Figure 2 13 4 each DISCSAT triggers a timing cycle that for each event e Operates the PSH cards to capture analog data for each parameter using the specified DISCSAT Extension to control the acquisition window Sequences the analog data from each active parameter to the A to D circuit Sequentially places the converted digital values on the backplane e Resets the PSH cards for the next event The ADC and PSH Control card 15 configured
86. 79 CFM 115 Vac 8 2 4 4 PN 4237003C 8 1 19 8 PARTS LISTS MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY Table 8 1 16 Pneumatics Continued Part Number Description Figure Item 5120109 Switch drip chamber 8 2 5 3 5120109 Switch waste filter 8 2 5 4 5120109 Switch system pressure 8 2 5 5 5120178 Manifold assembly 25 psi 8 2 5 6 6028142 Cable assembly sheath service 8 2 3 6 8 2 4 9 6211015 Actuator pilot 7 16 bore 8 2 5 8 8 2 18 14 23 8 2 19 1 6216129 Fitting hose barb T brass 0 187 i d 8 2 6 10 6232092 Fitting quick connect white 8 2 3 2 8 2 4 8 6232096 Valve solenoid 8 2 5 7 6232104 Fitting hose barb T 0 115 to 0 180 i d 8 2 3 17 8 2 6 5 6232309 Fitting hose quick connect panel mount blue 8 2 3 17 8 2 4 7 6232413 Fitting miniture nipple 10 32 8 2 5 8 6232417 Fitting hose barb 0 062 in i d PVC 8 2 6 6 6232418 Fitting hose barb 0 115 0 180 in i d PVC 8 2 6 4 6232449 Quick connect orange 6232450 Fitting panel mount bottle blue male 8 2 3 9 15 8 2 4 13 6232451 Fitting panel mount bottle white male 8 2 3 3 8 2 4 12 6232452 Fitting panel mount bottle yellow male 8 2 3 9 6232490 Regulator sample low bleed regulator 8 2 5 1 6232491 Regulator sheath low bleed regulator 8 2 5 2 Used with Pneumatic Interface Card PN 6705504 6704552 Pneumatic Interface Card Used only with Norgren pressure regulators 8 2 4 3 6801536 Valves check for 0 156 i d tubing was
87. 9 T fitting PN 6216129 9 Tubing 1 in 3 cm for end of new T fitting PN 3202039 9 Wire tie mount PN 6011017 4 Wire tie PN 6011001 8 Double sided tape PN 8023259 4 Metal strip PN 1021020 8 Autoclone Sorting Option mechanism screws PN 2839073 and washers PN 2827135 2 Deflection body assembly PN 6704311 1 Gloves Software version 4 01 or above is required Electrical and Mechanical Inspection 1 2 Verify that the 256 Memory card Cytometer CPU card has been installed Verify that the instrument has been upgraded to have the thin insertion body flow cell and tip with the 3000 Vdc modules Verify that there are no broken components on the Autoclone Sorting Option Interconnect and Autoclone Sorting Option Position cards Verify that there are no loose or damaged mechanical components Verify that Bio Hazard Upgrade Kit has been installed Verify that the Elite system sorts correctly Make corrections before proceeding Preliminary Procedure E S ek cB wA 11 12 13 Turn power switch OFE Remove the front right panel Data Acquisition Card cage cover Remove the Sort Oscillator card Replace R7 with a 4 99 kOhm resistor Replace R12 with a 10 kOhm resistor Install the Sort Oscillator card Remove the deflection body Remove the sample collection drawer Remove the existing 1 4 turn brackets from frame where sample collection drawer was fastened Remove the back pa
88. 9 23 3 10 1 Autoclone Sorting Option Mechanism 3 10 1 3 12 1 Laser Beam Hitting Dichroic Top View 3 12 2 3 13 1 Coax Cable Connections 3 13 4 3 14 1 Enterprise Laser Mounting Location 3 14 3 3 14 2 Beam Separator Location 3 14 3 3 19 1 Waste Bottle Caps 3 19 3 TABLES 3 1 1 Power Requirements 3 1 2 3 1 2 Water Specifications for Coherent 305 Laser 3 1 2 3 1 3 Environmental Requirements 3 1 3 3 1 4 Required Operating Clearance 3 1 4 3 7 1 Possible Laser Configurations 3 7 1 3 9 1 Gated Amp Jumper Configuration 3 9 4 3 9 2 Coax Cable Interconnections 3 9 5 3 9 3 PMT Adjustments 3 9 12 3 9 4 Offset Adjustments 3 9 18 3 10 1 Spade Lug Connections 3 10 5 3 11 1 Card and Cable Connections 3 11 3 3 13 1 Circuit Card Revision Level 3 13 2 3 13 2 Additional Information on Cards 3 13 2 3 13 3 Circuit Cards in ESP Equipped Units 3 13 5 3 21 1 Technical Specifications of the Pentium Computer 3 21 1 4237003 3 V 3 vi PN 4237003 INSTALLATION PROCEDURES PART A INSTRUMENT INSTALLATION WARNING Risk of personal injury Installation of an Elite system by anyone who has not successfully completed a Beckman Coulter authorized training course is prohibited Avoid personal injury by obtaining the necessary training to complete these installation procedures 3 1 PREINSTALLATION CHECK Before installing the instrument verify the following conditions Education Center tra
89. 91015 lt JU SHS 04NI ei T 20 WouJ Asng U9 3 san 3314009 9111 AE 4090 1 09 1N3A3 Ssod l Wad AN DO IN AWA puro sun 01 9030 Due 1591 o sindur sayejnwisg quei 3uvdWO02 91607 om pue oe 3110210 1391 HOS JIp DU 1104 i gt OU Io d ll ee m Sang Hsiuog 491 HOS pafejaq uos poKejoq pouad 990455 uos e Huvi viva ABOIN3IN jojdoJp 190 sawn p usd dyno 301711950 pajdwes geq HOS aqonss WOMJ 190801 1052910 NO OA STWNOIS 1119319 AV1S0 15411405 4 bech E 5 oc 189 os SS jesasip X909 POS m 940455 58 SUOISI299p HOS UE ouAs ui uj yos 49127 mm S 8 e ejag uos weeg 2019 6 9 z ant PN 4237003C INSTRUMENT DESC SORT SUBSYSTEM Sort Osc RIPTION illator Card Coincidence Logic The delayed sort information from the Sort Delay card comes into the Sort Oscillator card Figure 2 8 6 and is clocked into the card at quarter droplet intervals
90. ADJUSTMENT __ LAGER SHUTTER 7003196A BEAM SHAPER SCREW Optical Fine Tuning Optical fine tuning is achieved through the use of fluorescence peak pulses To properly optimize the system the pulses should originate from a top PMT reflected image such as PMT 2 and the last PMT straight through image If the optimal for one path is not optimal for the other the adjustment is positioned between the two optimal positions 1 At the Cytometer a Select Main b Adjust the PMT2 high voltage for a PMT2 peak pulse of about 5 V high See Figure 4 12 14 4 12 8 PN 4237003C 4237003 SERVICE AND REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE Figure 4 12 14 2 High Voltage Setting PMT2 PEAK 5 VOLTS B S 7003201A Ifthe PMT2 pulse does not appear or is very noisy the flow cell may not be aligned enough to the center of the fluorescence pickup lens Adjust the flow cell vertical and horizontal knobs until the pulse appears The traces are set differently depending on if the last is PMT4 or PMT5 gt Ifyou have 4 system go to PMT4 System gt Ifyou have a 5 system go to PMT5 System PMT4 System ATTENTION Risk of losing traces if the steps for fine tuning the PMT4 are not followed in exact sequence Follow the sequence of steps to avoid losing the traces when setting up the scope 1 At the Cytometer select a 1 and PMT2 Peak b
91. An easier method is to observe the pulses on the systems oscilloscope display and to estimate the time needed It is important that you allow for the slowest rising pulse integral and log With either method fine tune the setting by acquiring histograms of the desired parameters Then begin with the highest ASD reduce the time until you notice a shift in the population s mean channel At that point set the ASD to the next higher longer setting See Figure 2 13 7 for ASD settings 2 13 10 PN 4237003 INSTRUMENT DESCRIPTION 2 SIGNAL SELECTION AND DIGITIZATION Figure 2 13 7 ASD Settings Diagram Le bk em e e 7003039 DISCSAT DISCSAT EXT DISCSAT DISCSAT EXT 4 MICROSECONDS 15 MICROSECONDS 4MICROSECONDS 2 5 MICROSECONDS Poor ASD setting time is too Poor ASD setting time is too short long May allow noise or doublet Integral signal has not reached peak to contaminate signal value before window closes ADC and PSH Control Card The ADC and PSH Control card does not support true ASD operation During bootup the presence of this card is flagged by the message DISCSAT EXTENSION NOT PRESENT The Cytometers OPTIONS screen offers the following choices e FPW Full Pulse Width The acquisition window starts and stops at the discriminator crossings of the trigger channel This is the default setting and can be used in most cases if the trigger channel is an integral pulse If a peak signal is used as the disc
92. Connections 3 4 4 Laser Option 3 4 4 Optical Drive Option 3 4 4 Printer Option 3 4 4 3 5 TESTING ASSEMBLIES 3 5 1 Cytometer Control Computer 3 5 1 Compressor Module 3 5 2 Pneumatics 3 5 2 Camera 3 5 3 Lasers 3 5 4 Argon Laser 3 5 4 HeNe Laser 3 5 4 Optional Lasers 3 5 4 Computer 3 5 4 Alignment Check 3 5 4 3 6 SYSTEM TESTING 3 6 1 Acquisition Check 3 6 1 Analysis Check 3 6 1 Sort Check 3 6 1 Drive Frequency 3 6 3 Phase Adjustment 3 6 3 Delay Calculation 3 6 5 Monitoring the Sort 3 6 7 PN 4237003C 3 i 3 8 39 3 10 SAL PART B OPTIONS AND UPGRADES INSTALLATION OPTIONAL LASER INSTALLATION 3 7 1 Purpose 3 7 1 Tools Supplies Needed 3 7 1 Procedure 3 7 1 HOUSE AIR INSTALLATION 3 8 1 Procedure 3 8 1 GATED AMPLIFIER UPGRADE 3 9 1 Purpose 3 9 1 Tools Supplies Needed 3 9 1 Procedure 3 9 2 FALS Adjustment Assignment and Adjustment Potentiometer Location 3 9 10 PMTI Adjustment 3 9 12 AUX 1 Adjustment 3 9 16 AUX 2 Adjustment 3 9 16 PMT 2 Adjustment 3 9 17 PMT 3 Adjustment 3 9 17 PMT 4 and 5 Adjustment 3 9 18 Laser Alignment With Gated Amplifier 3 9 22 Laser Configuration 3 9 22 Laser Alignment 3 9 23 Calibration 3 9 24 AUTOCLONE SORTING OPTION INSTALLATION 3 10 1 Purpose 3 10 1 Tools Supplies Needed 3 10 1 Electrical and Mechanical Inspection 3 10 2 Preliminary Procedure 3 10 2 Upgrading the Instrument to be Autoclone Sorting Opt
93. Diagram J2 FL1 OUT Vote AII divider outputs are inverted Programmed Divider Outputs to be subtracted Programmed Divider Outputs to be subtracted 4 FL2 OUT Programmed Divider Outputs to be subtracted Programmed Divider Outputs be subtracted J7 PROGRAMMABLE FL3 PEAK OUT GAIN AMPLIFIER Programmed Divider Outputs to be subtracted INTEGRATOR 4 Programmed Divider 2 J6 Outputs to be E FL3 INTEGRAL subtracted P PROGRAMMABLE OUT GAIN AMPLIFIER CONTROL DECODE COMMANDS Dor dd J8 DIVIDERS FROM BACKPLANE CPU p E 4 FL3 LOG OUT TO SET AND CONTROL SET INTEGRAL LOG AMP SUBTRACTION CARD RUNGTIONS GAIN SET INTEGRATOR SPEED BANDWIDTH 7003055A P1 BACKPLANE CONNECTOR Dual FL SW R Card The Dual FL SW R card accepts two FL inputs and processes each input to provide variable gain peak and integral signals as well as a log output See Figure 2 10 2 PN 4237003C 2 10 1 2 INSTRUMENT DESCRIPTION CARD DESCRIPTION ALL CONFIGURATIONS Figure 2 10 2 Dual FL SW R Card Block Diagram Ji FL1 IN J5 FL2 IN PROG
94. E4 E5 E7 Scat CV SW R1 E9 E11 E5 E7 Sensor Interface 2 E5 E6 E8 E9 E11 E12 E13 E14 E16 E17 E19 E20 E22 E23 Data Acquisition Card Cage Table A 2 4 Data Acquisition Card Cage Jumpers Card Jumpers Pulse Generator and Clock R X1 through X4 Pulse Pileup Det TOF E1 E2 E20 E21 Mux and Scope Interface E1 E2 E3 E4 Quad PSH 1 E2 E4 Quad PSH 2 E2 E4 Peak ADC and PSH Control E1 E2 E6 E7 Data Lister Out E5 E6 Prism and Sort Window Test None Bitmap and Sort Decision None Interface and Scaler R E1 E5 E2 E6 E3 E7 Sort Delay R3 None PN 4237003C A 2 3 QUICK REFERENCE INFORMATION JUMPERS AND SWITCH SETTINGS Table A 2 4 Data Acquisition Card Cage Jumpers Continued Card Jumpers Sort Oscillator R2 None Sort Output R None Jumpers for Line Voltages Table A 2 5 Line Voltage Jumpers 100 Vac input to main system transformer TB9 2B to TB9 4B TB9 7B to TB9 9B TB9 1B to TB9 2B TB9 6B to TB9 7B jumper cable wae jumper cable jumper brass jumper brass 115 Vac input to main system transformer TB9 2B to TB9 4B TB9 3B to TB9 5B TB9 1B to TB9 2B TB9 5B to TB9 6B jumper cable wa jumper cable jumper brass jumper brass 230 Vac input to main system transformer TB9 1B to TB9 2B TB9 3B to TB9 4B TB9 5B to TB9 6B jumper brass jumper brass jumper brass 240
95. HOLVINDAY A 9 Quvo FHL A8 100 10d ATIVILNANDAS 38V 1N3A3 H9V3 303 SAN TWA YALA Wed JHL HIN VLVO 3NV IdX9v8 NO SINIT VIVO ANY SS38QQV 133319 3anirtaav 703109 3SV3U9NI 9p 1 pue 8seaJou 174110 LH0S Duruos 10 uomejeuo awy uu 35109 uos 1908 INH03 YIN Asnrav 3SvHd IEUUON 10 Pat page 19319 39341909 2101219099 1891 HOS LO 33151 0151934 TVNH A 3uVdIIO2 9 TOH 1NO2 dNWIUg dVIALIg c 703109 HSd 9 90V SIIONIN WSIdd ONY N0I938 YVANIT 4 9vg LNOUd Y01YTIS0 1HO S AN DO 1405 YVIVOS ANY 39v3831NI 101 241 waysAsqns yos weeg 2019 207 334 109 8 3301 WU 91907 09 1V LIIG JOWAYALNI 10 NOISI93Q 130 5111534 01938 YVANIT 1541 MOCNIM 130 134 2 8 3 PN 42370036 INSTRUMENT DESCRIPTION SORT SUBSYSTEM 2 8 4 The following text addresses the sorting process through the following circuit cards For more detailed information see the circuit schematics listed below
96. INTRODUCTION briefly describes this manual and provides essential safety information CHAPTER 2 INSTRUMENT DESCRIPTION introduces the Elite and Elite ESP and describes how each functions CHAPTER 3 INSTALLATION PROCEDURES includes procedures for installing the instrument and any options or upgrades CHAPTER 4 SERVICE AND REPAIR PROCEDURES includes procedures for servicing and repairing the instrument Each procedure includes a purpose and a list of tools and supplies needed to do the procedure CHAPTER 5 MAINTENANCE PROCEDURES includes procedures for maintaining the instrument CHAPTER 6 SCHEMATICS includes engineering schematics CHAPTER 7 TROUBLESHOOTING includes troubleshooting tables CHAPTER 8 ILLUSTRATED PARTS LIST includes the illustrated parts list APPENDIX A QUICK REFERENCE INFORMATION includes instrument tolerances and specifications jumper and switch settings test points and connectors APPENDIX B FILE PATHS provides a block diagram of the software file location Most of the material in this manual pertains to both the EPICS Elite flow cytometer and the EPICS Elite ESP flow cytometer In those cases where the information applies only to the EPICS Elite ESP flow cytometer that information is referred to as Elite ESP Numbering Format Each chapter of this manual is divided into topics that are numbered sequentially beginning at one The numbering format for the topic heading which is called the primary
97. LETTER is required Add if necessary Use the keyboard cursor keys to place the cursor to the far left of the page on a blank line and type PAPER LETTER Press Enter For A4 paper PAPER ISOA4 is required Add if necessary Use the keyboard cursor keys to place the cursor to the far left of the page on a blank line and type PAPER ISOA4 Press Enter ATTENTION The paper size to be used must not contain a REM statement before it 7 Verify that the CGI CFG file matches the following DRIVER C ELITE GSS GSSCGI SYS DRIVER C ELITE GSS META SYS G OUTPUT DRIVER C ELITE GSS HRVGA SYS G DISPLAY DISPLAY HRVGA SYS DRIVER C ELITE GSS MSMOUSE SYS G INPUT MOUSE MSMOUSE SYS MOUSEPORT COM1 FONTS C ELITE GSS rem standard laser printer driver rem DRIVER C ELITE GSS LASERJET SYS G PRINTER rem RESOLUTION 75 rem standard dot matrix printer driver rem DRIVER C ELITE GSS IBMGPR SYS G PRINTER rem RESOLUTION 75 rem HP Paintjet driver rem 90 dot per inch printer driver rem DRIVER C ELITE GSS HPPJ90 SYS G PRINTER rem Easy 2 users printer driver configuration rem DRIVER C ELITE GSS EPSONX SYS G PRINTER rem PRINTER EPSONX SYS rem EPSONX PRN rem RESOLUTION 120 DRIVER C ELITE GSS HPPCL5 SYS G PRINTER HPPCL5 PRN RESOLUTION 3 00 COLORS 16 FF ON ORIENTATION P
98. LOGIC 7003053A In Figure 2 8 3 FROM CPU INSTRUMENT DESCRIPTION SORT SUBSYSTEM TO BITMAP AND gt SORT DECISION CARD The digital words representing the channel number for each parameter are put on the Datapath bus one at a time The registers for levels provides storage for 8 sort regions Digital comparators compare the current parameter values to the stored regions The S strobe indicates the comparison is complete The DQUES are regions changed during sort The Control Logic circuit before sorting stores sort regions sent from the CPU in the registers During sort the appropriate regions are compared to the current parameter values Bitmap and Sort Decision Card Before a sort begins the system loads the Bitmap and Sort Decision card Figure 2 8 4 with the sort logic criteria which is the information the Workstation sends to the Cytometer The information contains the combination of gates and or bitmaps that must be met so that a cell is sorted to the right or to the left This card also contains the memory where the four available sort bitmaps are stored The system loads these memories before the sort begins The user draws an amorphous region on 2 8 5 2 INSTRUMENT DESCRIPTION SORT SUBSYSTEM 2 8 6 a two parameter histogram background Each X and Y channel number pair comprises a bit map location The Elite software assigns a bit map location a 1 value if it is wit
99. Light Sources 2 7 8 High Sensitivity Flow Cell Tips 2 7 8 SORT SUBSYSTEM 2 8 1 Function 2 8 1 Overview 2 8 1 Independent Operation 2 8 1 Sort Cycle 2 8 1 Circuit Cards 2 8 1 Prism and Sort Window Test Card 2 8 4 Bitmap and Sort Decision Card 2 8 5 Interface and Scaler Card 2 8 7 Sort Delay Card 2 8 8 Sort Oscillator Card 2 8 10 Function 2 8 11 Sort Output Card 2 8 13 Pulse Pileup Det TOF Card 2 8 15 TOF Function 2 8 16 PPU Function 2 8 17 Transistor Bracket Card 2 8 19 Power Supplies 2 8 19 PNEUMATICS SUBSYSTEM 2 9 1 General 2 9 1 Solenoid Valves 2 9 1 Circuit Cards 2 9 4 Pneumatic Interface Card 2 9 4 Sample Pressure Systems 2 9 4 Old Type 2 9 4 New Type 2 9 7 Sheath Pressure Systems 2 9 8 Old Type 2 9 8 New Type 2 9 10 Compressor Module 2 9 12 CARD DESCRIPTION ALL CONFIGURATIONS 2 10 1 3 PMT Sub SW R Card 2 10 1 Dual FL SW R Card 2 10 1 Scat CV SW R Card 2 10 2 Peak Scatter Mux SW R Card 2 10 3 CARD DESCRIPTION NON GATED AMP NON SWITCH CONFIGURATION EXCEPT ANALYZER 2 11 1 Scat CV Amp Card 2 11 1 2 iii 212 2 13 3 PMT Sub Amp Card 2 11 2 Function 2 11 2 Jumpers 2 11 2 Inputs 2 11 2 Dual FL Amp Card 2 11 3 Function 2 11 3 Inputs 2 11 3 CARD DESCRIPTION GATED AMPLIFIER CONFIGURATION 2 12 1 Gated Amp Control Card 2 12 1 Gated Amp R3 Card 2 12 2 Functions 2 12 2 Scat Aux Gated Amp Card 2 12 2 Functions 2 12 2 Inputs 2 12
100. MICROSECOND MICROSECOND MICROSECOND the 7 Microsecond DELAY LINE DELAY LINE DELAY LINE Delay card DECODE AND CONTROL oss SELECT 20 LT 40 OR 60 P1 m Card edge DC RESTORE DELAY OUT AND OFFSET 3 ADJUST J6 J9 J12 20 40 60 MICROSECOND DELAY CARD ONE OF FOUR CHANNELS SHOWN 7003023A 1 Microsecond Delay Card The 7 Microsecond Delay card delays the incoming pulse for seven microseconds Output gain and offset are adjustable See Figure 2 12 5 Figure 2 12 5 Block Diagram 7 Microsecond Delay Card INPUT AMPLIFIER 3 5 MICROSECONDS BUFFER DELAY 3 5 MICROSECONDS DELAY IN AMPLIFIER WITH J2 GAIN AND OFFSET OUT J4 e ADJUSTMENT J3 J5 J7 REF A N REF 7 MICROSECOND DELAY CARD ONE OF FOUR CHANNELS SHOWN EA 2 12 6 42370036 INSTRUMENT DESCRIPTION SIGNAL SELECTION AND DIGITIZATION 2 13 SIGNAL SELECTION AND DIGITIZATION Overview PN 4237003C Before considering the functions and operation of individual circuit cards you must first understand how the cards work together as a system to acquire data Each cell or particle that the Elite processes is called an event And each event represents one or more pulses containing different information about the same cell The Elite must process these pulses separately but retain the fact that the parameters are for the same cell Visualize the system as operating on an event by event basis or cycle The acq
101. OPTICS JET IN AIR 5mm OPTICS 7003186 42370036 2 3 14 INSTRUMENT DESCRIPTION 2 ELECTRONICS SUBSYSTEM 2 4 ELECTRONICS SUBSYSTEM Description Figure 2 7 1 is a block diagram of the Elite The left section consists of the Multibus card cage and peripheral devices Refer to Heading 2 5 CONTROL AND DISPLAY SUBSYSTEMS for more information The upper and lower card cages contain cards to support data acquisition sorting and fluidic control functions Refer to the appropriately named subsystem sections of this manual for more information Note The configuration of the upper backplane varies with the configuration of the instrument Refer to Heading 2 6 ACQUISITION SUBSYSTEM for more information 4237003 2 4 1 INSTRUMENT DESCRIPTION ELECTRONICS SUBSYSTEM 2 4 2 PN 4237003C INSTRUMENT DESCRIPTION 2 Figure 2 4 1 Block Diagram System Electronics
102. PARTS LISTS ILLUSTRATED PARTS Table 8 2 7 Dual CRT and Connector Panel Detail of Parts See Figure 8 2 6 Part Number Description Item Part Number Description 1 6856703 CRT assembly complete 11 6856950 Connector panel front CRT assembly without covers 2 6704441 KIR 3 card 12 2121586 Bail lock D type 3 6028045 Dual CRT cable 13 6028025 Data Lister Out INT cable 4 6027992 KIR 3 communication cable 14 6028021 Serial communication cable 5 4717714 Contrast control knob 15 2104261 Screw lock female 6 4717714 Brightness control knob 16 6028113 AUX video INT cable 7 4717714 Contrast control knob 17 6028068 Service INT cable 8 4717714 Brightness control knob 18 2105017 Duplex outlet 9 4717730 Dual CRT potentiometer trigger 19 6028070 BNC W31 cable adjustable 10 5120196 Scope trigger switch 1010165 Scope trigger knob PN 4237003C 8 2 7 8 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 8 Card Configuration Elite See Table 8 2 8 30 29 28 7003087A 1234567 8 91011121314 15 16 31 30 29 28 27 26 25242322212019 18 17 Table 8 2 8 Elite Card Configuration Detail of Parts See Figure 8 2 8 Part Number Description Item Part Number Description 1 N A Gated Amp card cage 16 16705117 Sensor Interface card 2 6704893 Gated Amp Control R2 card 17 6704201 Sort Output R card 3 6705249 Dual FL
103. REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE IMPORTANT Risk of misalignment if both the Collinear Laser Alignment and Gated Amp Laser Alignment are used on an instrument Select the appropriate procedure Do not do both the Collinear Laser Alignment Fine Tuning for Three Lasers and Gated Amp Laser Alignment Time Space Separated Laser Alignment on an instrument 24 Do one of the following e Laser Alignment Fine Tuning for Three Lasers e Gated Amp Laser Alignment Time Space Separated Laser Alignment Collinear Laser Alignment Fine Tuning for Three Lasers 1 Remove the targets a Remove the two standard 5 in optical bench mounted laser alignment targets b Remove the two standard vertical optical plate mounted targets Install the following a Scatter sensor assembly b Beam shaping assembly c Laser shutter Lower the flow cell to its proper position using the flow cell Y vertical adjustment knob and the camera Remove optical filters except the 1 0 ND filter from the forward scatter sensor filter holder and install the filter holder back in the FALS sensor Note Remove the 525 BP filter to obtain a side scatter fluorescent signal in PMT2 Press SHEATH to begin sheath flow At the Cytometer s Scope screen a Select Fals Peak for the lower trace b Select PMT 2 Peak for upper trace Note Throughout this procedure Fals Peak and PMT 2 are used for alignment purposes Op
104. S to enter the Setup screen Type P and press to select PRINTER CONFIGURATION Use 4 to highlight Interrupt row and press Spacebar Use 4 to highlight NO INTERRUPT and press Ente Use to highlight Output Method row and press Spacebar Use 4 to highlight Use BIOS to OUTPUT to Printer Plotter and press Enter Press to return to the PrintQ Screen Press to enter the Setup screen Type M to select Manage Profiles and press Ente In the Print When box select Start printing when capture is complete Note Do not change anything else on this screen Press to access the Profile Selections Dialog Page screen Use 4 to highlight Eject page at end row and press Spacebar Use to highlight Do not eject partial last page and press Ente Press to save changes Press to exit PrintQ program PrintQ is now installed Press att Delete to activate changes and to reboot the Workstation At the Protocol screen verify CYTOSETTINGS SEND Press F9 If ALL DISCRIMINATOR SET TO OFE CONTINUE WITH START Y N appears press Y J Enter PN 4237003C SERVICE AND REPAIR PROCEDURES 4 SOFTWARE INSTALLATION 50 If the Cytometer program needs to be downloaded UNABLE TO RECEIVE CYTOMETER PROGRAM VERSION appears on the screen To download the Cytometer program a Press Enter b The question Do You Want to Transfer the Control Files appears Press Y Ente The Cytometer Touch Scr
105. TROUBLESHOOTING RESETTING CYTOMETER SOFTWARE LOCKUPS 7 6 2 4237003C PN 4237003C CONTENTS ra 8 PARTS LISTS 8 1 1 8 1 MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY 8 1 1 8 2 ILLUSTRATED PARTS 8 2 1 ILLUSTRATIONS 8 2 1 8 2 2 8 2 3 8 2 4 8 2 5 8 2 6 8 2 7 8 2 8 8 2 9 8 2 10 8 2 11 8 2 12 8 2 13 8 2 14 8 2 15 8 2 16 8 2 17 8 2 18 8 2 19 8 2 20 8 2 21 8 2 22 8 2 23 8 2 24 8 2 25 8 2 26 8 2 27 8 2 28 8 2 29 8 2 30 8 2 31 8 2 32 8 2 33 8 2 34 8 2 35 8 2 36 8 2 37 8 2 38 TABLES 8 1 1 8 1 2 Elite Cytometer Right Panel Removed See Table 8 2 1 8 2 1 Workstation See Table 8 2 2 8 2 2 Sheath and Waste Compartment See Table 8 2 3 8 2 3 Pneumatics Cabinet See Table 8 2 4 8 2 4 Pneumatics Drawer See Table 8 2 5 8 2 5 Pneumatics Panel Rear View See Table 8 2 6 8 2 6 Dual CRT and Connector Panel See Table 8 2 7 8 2 7 Card Configuration Elite See Table 8 2 8 8 2 8 Card Configuration Elite ESP See Table 8 2 9 8 2 9 Cable Interconnections Elite and Elite Analyzer See Table 8 2 10 8 2 10 Multibus Card Cage All Configurations See Table 8 2 11 8 2 11 Data Acquisition Card Cage Elite ESP See Table 8 2 12 8 2 12 Cytometer Inside Rear View See Table 8 2 13 8 2 13 Cytometer Right Rear Door Inside View See Table 8 2 14 8 2 14 Cytometer Left Rear Door Inside View See Table 8 2 15 8 2 15 Cytometer Left Front Corner Inside View See Tabl
106. Tahle 8 2 21 Deflection Body Detail of Parts See Figure 8 2 21 Item Part Number Description 1 6704754 Deflection body and holder 2 N A Deflection body holder 3 N A Deflection body PN 4237003 8 2 21 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 22 Photomultiplier Tubes PMTs See Table 8 2 22 S 7003092 2 Sch Table 8 2 22 Photomultiplier Tubes PMTs Detail of Parts See Figure 8 2 22 Item Part Number Description 1 6856505 PMT 4 2 6856833 PMT 5 3 6856833 PMT 3 4 6856833 PMT2 5 6856833 1 8 2 22 42370036 PARTS LISTS 8 ILLUSTRATED PARTS Figure 8 2 23 Autoclone Sorting Option Mechanism See Table 8 2 23 HSVHd Z p RENI A Ji NEP Na 7003146A Table 8 2 23 Autoclone Sorting Option Mechanism Detail of Parts See Figure 8 2 23 Item Part Number Description 1 6705328 Interconnect card 2 6028266 Flex cable 3 6028266 Main cable 4 3501018 Stepper motor 5 2508022 Shaft coupling 0 188 and 0 250 i d ends 6 5103013 Snap action switch 7 4836701 Opto switch 8 6705329 Position Detection card 9 4818006 Opto switch 0 1 gap 10 6858187 Tray support 11 1018721 Tray cover 12 1020199 Tray 13 3505035 Stepper motor 1 8 degree 5 5 Vdc PN 4237003C 8 2 23 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 24 Autoclone Sorting Option Beak Waste Catcher See Table 8 2 24
107. UPGRADE 3 22 1 Purpose 3 22 1 Tools Supplies Needed 3 22 1 Procedure 3 22 1 Install the New Bitmap and Sort Decision Card 3 22 1 Install The Elite Software 3 22 1 SERVICE AND REPAIR PROCEDURES 4 1 1 4 1 OPTICAL ALIGNMENT PROCEDURE 4 1 1 System Alignment 4 1 1 Purpose 4 1 1 Standard Configuration Procedure 4 1 1 Tools Supplies Needed 4 1 1 Procedure 4 1 1 Standard Laser Alignment 4 1 2 Purpose 4 1 2 Tools Supplies Needed 4 1 2 Procedure 4 1 2 Flow Cell Alignment Initial 4 1 3 PN 4237003C PN 42370036 42 4 3 4 4 Beam Translator Equipped System Alignment 4 1 3 Purpose 4 1 3 Tools Supplies Needed 4 1 3 Laser Mounting Procedure 4 1 4 Laser Alignment for Gated Amp Systems Standard Laser Alignment 4 1 4 Collinear Laser Alignment Fine Tuning for Three Lasers 4 1 6 Gated Amp Laser Alignment Time Space Separated Laser Alignment 4 1 7 Camera Adjustment Procedure 4 1 9 Aiming the Camera 4 1 9 Focusing the Camera 4 1 10 Zooming the Camera 4 1 10 Positioning Movement of the Camera 4 1 11 Optimizing the Image 4 1 11 SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE 4 2 1 Purpose 4 2 1 Tools Supplies Needed 4 2 2 Crystal Drive Test Procedure 4 2 2 Crystal Drive Adjustment Procedure 4 2 2 Sort Pulse Amplitude Test 4 2 2 Sort Pulse Amplitude Adjustment 4 2 3 Phase Settings 4 2 3 Delay Settings 4 2 3 Sync Adjustment 4 2 4 Sort Function Verificati
108. Units 3 13 5 Technical Specifications of the DTX Pentium Computer 3 21 1 Pulse Generator Card Values 4 2 15 Line Voltage Jumpers 4 3 1 Power Supply Measurement and Adjustments 4 4 2 Potentiometer Adjustments 4 5 1 Back Panel Wiring Configuration 4 10 1 Multibus Card Cage Cards Jumper and Switch Settings 4 10 1 Non Gated Amp Card Cage Jumper Setting 4 10 2 Gated Amp Card Cage Jumper Settings 4 10 3 Data Acquisition Card Cage Cards Jumper Settings 4 10 3 Cable Connections 4 10 4 Cable Connectors 4 10 10 Isolating Problem Areas 7 1 1 Autoclone Sorting Option General Troubleshooting Information 7 1 2 General Troubleshooting Information 7 1 3 Lasers General Troubleshooting Information 7 1 5 Optics General Troubleshooting Information 7 1 5 Electronics General Troubleshooting Information 7 1 6 Software General Troubleshooting Information 7 1 6 Power Supplies General Troubleshooting Information 7 1 6 Fluidic Components 7 3 1 Troubleshooting Sort Problems 7 5 2 Autoclone Sorting Option 8 1 1 xxiii xxiv 8 1 2 8 1 3 8 1 4 8 1 5 8 1 6 8 1 7 8 1 8 8 1 9 8 1 10 8 1 11 8 1 12 8 1 13 8 1 14 8 1 15 8 1 16 8 1 17 8 1 18 8 1 19 8 1 20 8 1 21 8 1 22 8 1 23 8 2 1 8 2 2 8 2 3 8 2 4 8 2 5 8 2 6 8 2 7 8 2 8 8 2 9 8 2 10 8 2 11 8 2 12 8 2 13 8 2 14 8 2 15 8 2 16 8 2 17 8 2 18 8 2 19 8 2 20 8 2 21 8 2 22 8 2 23 Beam Shapers 8 1 2 Cables 8 1 2 Card
109. Vac input to main system transformer EE a ee e TB9 1B to TB9 2B jumper brass TB9 4B to TB9 8B jumper cable TB9 6B to TB9 10B jumper cable 100 Vac input to laser auto transformer TB10 1B to TB10 4B jumper cable TB10 2B to TB10 8B jumper cable TB10 6B to TB10 9B jumper cable TB10 8B to TB10 10B jumper cable TB10 4B to TB10 5B jumper brass TB10 6B to TB10 7B jumper brass Remove TB10 1B to TB10 10B jumper cable 115 Vac input to laser auto transformer TB10 1B to TB10 10B jumper cable 230 Vac input to laser auto transformer TB10 1B to TB10 4B jumper cable TB10 6B to TB10 9B jumper cable TB10 8B to TB10 10B jumper cable TB10 7B to TB10 8B jumper brass Remove TB10 1B to TB10 10B jumper cable 240 Vac input to laser auto transformer TB10 1B to TB10 3B jumper cable TB10 6B to TB10 9B jumper cable TB10 8B to TB10 10B jumper cable TB10 7B to TB10 8B Jumper brass Remove TB10 1B to TB10 10B jumper cable A 2 4 PN 4237003C QUICK REFERENCE INFORMATION A OPTICAL SETTINGS OPTICAL SETTINGS Collection Optics Upgrade Crystal drive 0 096 Deflection 0 096 Sort Waveform Verification Drop frequency 32 kHZ so each droplet period is 31 25 microseconds Crystal drive 100 Deflection amplitude 10096 Sort Test ON Sort Right Enable ON Sort Left Enable ON Stream control sort pulse baseline 0 V Front Porch lengt
110. Vdc on orange wire 15 0 0 05 Vdc on green wire Both referenced to black wire 15 V 5 0 A supply Locate the Pulse Gen amp Clock card in the Data Acquisition lower backplane far left slot and adjust the larger 15 Vdc Analog Power Supply using a DVM as follows Black DVM Lead Red DVM Lead DVM Result Black test point Orange test point 15 0 V 0 05 Black test point Green test point 15 0 V 0 05 15V 3 4 A supply Adjust the 15 Vdc 3 A power supply connected to Bertans with DVM connected to the HV backplane at P13 on top Bertans backplane Put the black DVM lead on the black white wire Place the red DVM lead on the green wire and adjust for 15 1 V 0 05 Vdc Move the red DVM lead to the orange wire Adjust for 15 1 V 0 05 Vdc 90 V power supply Adjust the power supply using the DVM as follows Black DVM Lead Red DVM Lead DVM Result OUT Upper Supply OUT Upper Supply 90 Vdc 2 Vdc OUT Lower Supply OUT Lower Supply 90 Vdc 2 Deflection power supply Not adjustable Test at test points J2 black ground J3 blue 2 3 to 3 0 Vdc J4 red 2 3 to 3 0 Vdc 4 4 2 PN 4237003C 4 5 SERVICE AND REPAIR PROCEDURES PNEUMATICS SYSTEM PNEUMATICS SYSTEM ATTENTION This section contains procedures for two types of systems those with low bleed regulators and those with non low bleed regulators Do not do both procedures on an instrument Low B
111. a Press SHEATH b Verify Sort Pulse Enabled is off c Verify Deflection is 0 0096 d Disconnect the clip from the flow cell introduction rod and connect a multimeter in series to measure the current See Figure 4 11 2 Figure 4 11 2 Connecting Clip from Flow Cell to DVM SORT PULSE ALLIGATOR CLIP ONNECTION 9 FLOW CELL M INSERTION ROD 7003187A e Turn the stream deflection knob located on the sample station next to the mixer speed control all the way right or left f Locate the incorrect cracked or leaking fitting 1 Determine if there is current by increasing Deflection and checking your meter for any current indication 2 Ifcurrent is detected troubleshoot by pinching the tubing carrying the fluid to and from the flow cell until the current reading drops g Replace the defective fitting 3 Decontaminate the system a Remove the 0 2 m sheath filter and bypass it with a piece of tubing equipped with the correct fittings b Fillthe sheath tank with the 5096 bleach solution PN 4237003C 4 11 3 SERVICE AND REPAIR PROCEDURES SORT SENSE FLOW CELL ADJUSTMENT AND VERIFICATION 4 11 4 c Replace the flow cell with the biohazard flow cell included in the customers accessories box d Allow the system to run in sheath until the sheath tank is about one quarter full Note This should take about 45 minutes e Rinse the sheath tank thoroughly and fill with deionized water f Allow the syste
112. as needed Vacuum leak or obstruction Locate and repair Waste catcher does not extend Corroded cylinder Replace cylinder No pressure to cylinder Check tubing and solenoid valves Replace as needed PN 4237003C TROUBLESHOOTING ISOLATING THE PROBLEM AREA Table 7 1 3 General Troubleshooting Information Continued Problem Probable Cause Corrective Action Incorrect number of beads in well Host instrument not correctly adjusted for optimal sorting Do Sort Setup procedure in Operator s Guide Sorted stream is misaligned 1 Adjust stream alignment and deflection so sorted stream does not strike water catcher 2 Ensure plates are clean and dry Table 7 1 4 Lasers General Troubleshooting Information Problem Probable Cause Corrective Action Laser fails to turn ON Interlock keys Laser current is high on Cytometer Control Screen Dirty air intake filter Clean air intake filter Table 7 1 5 Optics General Troubleshooting Information Problem Probable Corrective Action Poor CVs with DNA Check or IMMUNO CHECK beads Optics are not aligned Do Heading 4 1 OPTICAL ALIGNMENT PROCEDURE Shifted mean channels Optics are not aligned Do Heading 4 1 OPTICAL ALIGNMENT PROCEDURE No data Optics are not aligned Do Heading 4 1 OPTICAL ALIGNMENT PROCEDURE No pulses appear or
113. by two mounting holes Note Leave enough room for yourself for a good view of the alignment holes in the targets 4 Remove these components a Scatter sensor b Laser shutter c Beam shaper 5 Raise the flow cell up as far as possible using the flow cell vertical adjustment knob 4237003C PN 42370036 10 11 12 13 14 15 16 SERVICE AND REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE Install the two smaller laser alignment targets on the vertical optical plate At the Cytometer Control screen a Turn the Hehe laser on b the air cooled Argon lasers Turn the UV laser on using lasers own on off switch Close the air cooled Argon and the UV laser output shutters to prevent any beam over exposure accidents With only the HeNe emitting align the HeNe using the Standard Alignment Laser procedure first through the two targets on the optical bench then through the two vertical optical plate mounted targets using the 90 degree dichroic Close the shutter on the HeNe laser Move the two optical bench mounted Targets in front of the air cooled Argon laser Open the shutter on the air cooled Argon aser Align the Argon Laser using the Standard Alignment Laser procedure for the two 5 in targets on the optical bench then for the vertical optical plate mounted targets using the 90 degree dichroic Close the shutter on the air cooled Argon laser Move the two optical bench mounted 5 in targ
114. c Select PMT3 UPBM d Adjust R111 slot 8 to minimize e Change the delay to 60 microseconds and the clock to 5 f Adjust R108 slot 7 to minimize g Change the delay to 7 microseconds move the blank to the bandpass position under PMT4 and change the clock to 0 5 Repeat steps 33 through 36 for PMT 4 and 5 if installed for the offset adjustments using Table 3 9 4 Use Align LED and 550 DCLP and mirror With PMT1 PEAK still selected as the trigger source set the pulse delay for 7 microseconds and set the clock to 0 5 microseconds Adjust the window width for 6 microseconds Adjust the window delay for 6 microseconds Ensure the gated amp is disabled Set Signal delays for UPPBM At the Scope screen a Select trace 1 for TRIG b Select trace 2 for PMT2 LOG Ensure optical filters are arranged to allow signals in both PMTs used as trigger and being tested Use a dichroic At the Main screen adjust PMT2 HV for a 1 division tall log pulse 3 9 21 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE 48 49 50 51 52 53 At the Gated Amp Control screen set GATED AMP ENABLE Note While monitoring PMT2 on trace 2 the amplitude should remain 1 division tall and should not increase or decrease when the gated amp is enabled or disabled If the signal changes in amplitude adjust R106 in slot 8 Quad 7 Microsecond Delay card until no change occurs See Table 3 9 4 for Sensor VS Off
115. card 10 Return E2 E3 to E1 E2 and readjust pneumatics as needed using Pneumatic Interface Card Adjustment Sample Pressure Problems This system is simple to troubleshoot if you remember that all the Pneumatic Interface card does is send a voltage to the regulator which represents the desired pressure The regulator does all the work it contains a pressure sensor and controls to the pressure to match the requested pressure The sample pressure sensor on the card is only used to obtain a pressure value to display on the Cytometer screen Connect your pressure gauge at the sample regulator output Connect your DVM to TP8 Set Sample Pressure to 15 psi apr You should read 10 on your meter If not do steps 1 through 4 of Low Bleed Regulator Adjustment Procedure for Sample Regulator under Heading 4 5 PNEUMATICS SYSTEM 5 Ifyou cannot obtain the correct reading in step 4 the Pneumatic Interface card is defective or the card is not being correctly set by the CPU Replace the card or inspect replace the ribbon cable to Sensor Card If you obtained the correct reading in step 4 the regulator is being set for 15 psi Measure the air pressure at the regulator output e fitis 15 psi the regulator is correctly set e Ifthe pressure is not correct calibrate according to Pneumatic Interface Card Adjustment under Heading 4 5 procedure If the correct pressure cannot be achieved and correct pressure is available to the
116. card SW R cards for compensation FL4 Not present Routed to 3 PMT Sub Not present SW R2 card for compensation Scat CV Amp Processes FALS and 90LS Processes the signal from J12 to card signals provide the integral and log FALS signals Mux and Scope card Receives all processed signals Receives all processed signals Peak Scatter Mux SW R card Not present Not present Receives FALS signal J11 branches and is routed off card at J12 Processes FALS peak signal and routes to Mux and Scope card The system cannot compensate for an F1 and F4 combination because they are not routed to a common card This is not a problem however because the two PMTs normally use optical filters with passbands that are far enough apart to prevent any spectral overlap peak scatter sensor is the FALS sensor in this system and is also used in the Gated Amp system 4 PMTs Nonswitchable Amps The 4PMT Nonswitchable Amp Figure 2 6 8 is the earliest configuration of the Elite With this setup PN 4237003C All fluorescence PMT signals go to the 3 PMT Sub Amp card Coax cables take FL1 and FL2 to the Dual FL Amp card for linear and log amplification The 3 PMT Sub Amp card completes the FL3 signal processing Coax cables route all processed signals to the Mux and Scope card The Scat CV Amp card processes all FALS and 90LS signals 2 6 9 2 oz E amp Go gt SES gt an
117. defined 4 5 1 sheath pressure system new type See sheath pressure system new type sheath pressure system old type See sheath pressure system old type sheath regulator adjustment 4 5 3 solenoid valves See solenoid valves test procedures 3 5 2 valves See solenoid valves Pneumatics Control card function 2 5 8 PNs See part numbers porch adjustments sorting requirements 3 6 5 Position Detection card function 2 2 3 sensors optical function of 2 2 3 switches mechanical function of 2 2 3 positive lens description 2 7 2 potentiometers Gated Amp adjustments 3 9 10 low bleed regulator adjustment 4 5 1 non low bleed regulator adjustment 4 5 4 speed control Motor Control card 2 5 8 power supplies 5 V function 2 8 19 12 V function 2 8 19 15 0 25A function 2 8 19 15 V 3 4A function 2 8 19 15 V 5A function 2 8 19 24 V function 2 8 19 36 V function 2 8 19 90 V 2 function 2 8 19 3000 V Deflection function 2 8 19 adjustment procedure 4 4 1 4 4 2 Bertans function 2 5 8 deflection function of 4 4 2 A 3 2 dimensions 3 2 2 fiber optics adjustments and measurements 4 4 2 A 3 2 location illustration 4 4 1 measurement procedure 4 4 1 4 4 2 switcher adjustments and measurements 4 4 2 A 3 2 troubleshooting 7 1 6 pressure regulators electronic adjustment of 4 6 1 Norgren when used 2 9 4 pressure switch adjustment 4 7 1 PN 4237003C pressures defined See pneumatics preven
118. double acting pinch valve When deactivated it vents the pressure in the sample tube to the drip chamber VL5 Flow Cell Vacuum Solenoid operated Applies vacuum to the flow cell Solenoid must be single acting pinch valve energized for pinch valve to close the vacuum tubing VL6 Sheath Pressure Solenoid operated Passes regulated sheath pressure to the sheath tank double acting pinch valve When deactivated it vents pressure in the sheath tank to the drip chamber VL7 Bubble Drain Solenoid operated Applies vacuum to the top of the bubble trap to remove single acting pinch valve any trapped air from the flow cell s path VL8 Rinse Pressure Solenoid operated Passes regulated sheath pressure to the rinse tank When double acting pinch valve deactivated it vents pressure in the rinse tank to the drip chamber VL9 Sheath Flow Solenoid operated Allows sheath to flow into the bubble trap because the single acting pinch valve pinch tube in the valve is released VL10 Sample Collect Solenoid operated Allows waste to flow to the bottle when the tube Drain double acting pinch valve connecting the drain to the waste bottled in unpinched VL11 Rinse Flow Solenoid operated Allows rinse liquid to flow into the bubble trap when the single acting pinch valve pinch tube in the valve is released VL12 Drip Chamber Drain Solenoid operated Applies vacuum to the drip chamber for draining when double acting pinch valve deactivated VL13 Autoclone Sortin
119. from leaving the instrument Use a white piece of paper Such as a business card to locate the beam during alignment 16 Use the tall targets to align the UV laser to the optical table 17 Secure the laser in place with the clamps provided 18 Remove the tall targets 19 Install the UV mirror assembly on the rail Use the mirror adjustments to target the UV laser through the small targets 20 Install the beam expander Figure 3 7 11 between the UV laser and the air cooled Argon laser and align it with the small targets The beam entering the expander will be about 0 3 mm in diameter The beam leaving the expander will be about 0 15 mm in diameter just before it passes through the HeNe mirror 3 7 10 4237003 PN 42370036 INSTALLATION PROCEDURES OPTIONAL LASER INSTALLATION Figure 3 7 11 Beam Expander Reducer VERTICAL TRANSLATION x LATERAL TRANSLATION FOCUS RING HORIZONTAL TRANSLATION HORIZONTAL ROTATION KNOB ERTICAL FINE OTATION HEIGHT SETSCREW MICRO FINE ee ADJUSTMENT SETSCREW AIL SETSCREW SE 21 Remove targets and replace the FALS sensor flow cell tip and shutter 22 Use the camera to align the flow cell 23 Run fluorospheres 24 Align the beam shaper for optimal signals on FALS and PMT 4 with the HeNe Note Remove the 675 BP and the ND filter On 5 PMT systems also remove the 640 DL 25 Block the HeNe beam to allow the air cooled Argon beam
120. functions you need to simulate two cells passing through the laser beam close together The test pulse generator is used in the dual pulse mode to simulate two cells You can vary the spacing delay between the pulses to simulate the spacing between the cells 4 2 9 4 SERVICE AND REPAIR PROCEDURES SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE 4 2 10 Tools Supplies Needed Oscilloscope Setup Pulse Generator 1 Use your oscilloscope to monitor J2 on the Pulse Generator card 2 Goto the Cytometer Service screen access code 26 28 32 and select Debug 2 3 If Mode does not display Single press Mode until it does 4 Press the following blocks on the touchscreen PORT 0 2 0 2 OUTPUT D 6 0 1 START PORT 0 2 0 4 OUTPUT F F F F START 5 Observe the pulses on your oscilloscope Ifyou do not observe pulses repeat step 4 e Ifyou do not observe a pair of pulses from 10 to 100 microseconds apart with a one millisecond spacing between pulse pairs perform the Pulse Generator Card Adjustment procedure in this section Ifyou observe pulses go to step 6 6 Press PORT 0 2 0 4 OUTPUT 0 0 1 0 START Note The observed pulse should be less than 1 V in amplitude Adjust the OUTPUT value in the command so the pulse is between 200 mV and 1 V FFFF max 0000 min Locate and remove the signal cable coming from PMT2 going to Acquisition electronics Connect a coax cable from J2 on the Pulse Generator card t
121. guidelines to follow 7 1 3 improper functions 7 3 1 leaks 7 3 1 log amplifiers 7 2 4 optics 7 1 5 3 PMT 2 compensation 7 2 3 pneumatics 7 5 1 power supplies 7 1 6 pressure controls problems 7 4 1 problems how to isolate 7 1 1 sample pressure problems 7 4 1 sheath air leaks 7 3 2 sheath liquid leaks 7 3 2 sheath pressure problems 7 4 1 sort current leakage test use of 7 5 1 7 6 1 sorting problems 7 5 1 7 6 1 tubes placement during sorting 3 6 2 tubing connections sheath and rinse bottles 3 4 3 sheath and waste bottle illustration of 3 4 4 PN 4237003C INDEX U unload the instrument 3 2 2 updated information See Notice of Information Update form V Vac inputs See inputs Vac vacuum increased causes of 2 9 1 valve assembly bracket See fittings valves solenoid See solenoid valves verification inspection procedure See VIP video output of camera See camera Video subsystem block diagram 2 5 4 description 2 5 4 VIP operational test electronics 5 1 1 operational test fluidics 5 1 1 operational test optics 5 1 1 operational test sorting 5 1 2 operational tests for every call 5 1 2 service not done in past 30 days 5 1 1 TOF verification procedure 5 1 2 what to do on every call 5 1 1 when to perform 5 1 1 VL1 function 2 9 2 VL2 function 2 9 2 VL3 function 2 9 2 VL4 function 2 9 2 VL5 function 2 9 2 VL6 function 2 9 2 VL 7 function 2 9 2 VL8 fun
122. heading is chapter number decimal point topic number For example the primary heading number for the fifth topic covered in Chapter 4 is 4 5 The page figure and table numbers are linked directly to the primary heading number For example Heading 4 5 begins on page 4 5 1 the first figure under Heading 4 5 is Figure 4 5 1 and the first table under Heading 4 5 is Table 4 5 1 Note Primary headings always begin on the top of a right page 4237003C PN 42370036 INTRODUCTION MANUAL DESCRIPTION Special Headings Throughout this manual WARNING CAUTION IMPORTANT ATTENTION and Note headings indicate potentially hazardous situations and important or helpful information Warning WARNING indicates a situation or procedure that if ignored can cause serious personal injury The word WARNING is in bold faced text in the printed manual and is red in the online manual Caution A CAUTION indicates a situation or procedure that if ignored can cause damage to equipment The word CAUTION is in bold faced text in the printed manual and is red in the online manual Important An IMPORTANT indicates a situation or procedure that if ignored can cause erroneous test results The word IMPORTANT is in bold faced text in the printed manual and is red in the online manual Attention An ATTENTION contains information that is critical for the successful completion of a procedure and or operation of the instrument The word
123. instrument for ready reference Copies are available from American National Standards Institute 1430 Broadway New York NY 10018 WARNING Risk of personal injury The laser beam can cause eye damage if viewed either directly or indirectly from reflective surfaces such as a mirror or shiny metal surface To prevent eye damage avoid direct exposure to the beam Do not view it directly or with optical instruments except with special service tools as directed in this manual Eye and skin damage as well as instrument damage can be caused by the laser beams The laser has enough power to ignite substances placed in the beam path even at a distance Indirect contact with the laser beam from reflective surfaces such as jewelry called specular reflection might also cause damage Follow these precautions e Never look directly into the laser light source or at scattered laser light from any reflective surface Never look down into the beams source e As a precaution against accidental exposure to the output beam or its reflection personnel performing service or maintenance procedures on the system should wear proper laser safety glasses Do not use lasers in the presence of flammables or explosives these include volatile substances such as alcohol solvents and ether Avoid direct exposure and indirect reflection of the laser beam to your skin e Assure that any spectators are not potentially exposed to a hazardous condition
124. laser beam on the flow channel Do not rotate the flow cell in this step c Tighten bolts 8 For systems with the newer enclosed bimorph adjust the thumbscrew to center the laser beam on the flow channel 9 Observe reflected laser beam and path of beam through the flow cell Repeat above adjustments until reflections are orthogonal and beam is centered on channel Beam Translator Equipped System Alignment Purpose Beam translators are available as upgrade parts They are not part of the modification kit for the HeCd laser but are offered as a sales option This translator is used to adjust the beam to the proper height The resultant beam is parallel to the exit beam of the laser This allows the user to use either co lateral or separated beams depending on the application Tools Supplies Needed Beam Translator Kit PN 6912985 4 which includes Beam translators one for each laser e UV light either HeCd or Argon UV laser PN 6858244 0 488 nm light Argon laser PN 6858245 8 4 SERVICE AND REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE 633 nm light HeNe laser PN 6858243 1 3micro optical rails for mounting PN 3814206 2 2 machine screws for each of above PN 2851763 7 New laser alignment targets with three target holes Large targets 6857274 6 Small targets PN 6857236 3 Mounting screws PN 2851920 6 Fluorospheres Laser Mounting Procedure Note The long dimension of th
125. liquid pressure Test this by raising or lowering the sheath bottle You will hear the regulator cycle and notice the sheath air pressure change to compensate for the liquid height change Figure 7 4 1 Troubleshooting Fluidics for No Sheath Flow Fluidics logically troubleshooting a typical problem No sheath flow No Sheath Flow Is Sheath Pressure Present No Yes as indicated on Cytometer screen 0 Is the Sheath pinch valve No Sheath Pressure Tree VL 9 opening No Do other valves operate es Is the pinch tubing stuck closed LISS or clogged Measure power supply No voltages at Pneumatic Intfc Card connection Replace tubing Are voltages correct No Yes Turn Sheath OFF No Is voltage 24V present Yes Remove sheath line from flow cell Repair Power on solenoid Momentarily turn Sheath On Does liguid spout from the disconnected tube No Yes Supply 7003082A Check cable connections Check Wiring between Replace Solenoid Replace Sheath Unclog or Repllace Pneumatic Solenoid and Pneumatic Filter Replace Flow Intfc Card Infc Card Cell Tip 7 4 1 7 TROUBLESHOOTING TROUBLESHOOTING PRESSURE CONTROL PROBLEMS 7 4 2 Move the jumper on the Pneumatic Interface card from 1 E2
126. match the printer currently in use if any Deactivate the active printer driver a Use the keyboard cursor keys to place the cursor to the far left of the page on the same line as the driver b Type REM and press Spacebar c If the active printer driver is for a Laserjet a standard dot matrix or an Epson printer 1 Use the keyboard cursor keys to place the cursor to the far left of the page on the Resolution line 2 Type REM and press d Ifthe active printer driver is an Epson printer 1 Use the keyboard cursor keys to place the cursor to the far left of the page on the Printer Epsonx sys line 2 Type REM and press 3 Use the keyboard cursor keys to place the cursor to the far left of the page the Epsonx Prn line 4 Type REM and press Use the keyboard cursor keys to place the cursor to the far left of the page on a blank line and a DRIVER C ELITE GSS HPPCL5 SYS G PRINTER and press b Type HPPCL5 PRN and press Enter c Type RESOLUTION 300 and press Enter d Type COLORS 16 and press Enter e FF ON If FF OFF appears without a REM statement before it a Use the keyboard cursor keys to place the cursor to the far left of the page on the FF OFF line b Type REM and press Spacebar PN 4237003C INSTALLATION PROCEDURES 3 COLOR PRINTER UPGRADE 6 Verify that the CGI CFG is setup correctly for the desired paper size e For 8 5 x 11 in paper PAPER
127. means This is normal Do Sort Function Verification under Heading 4 2 3 13 3 INSTALLATION PROCEDURES SORT PERFORMANCE ESP UPGRADE 3 13 4 15 Sort with beads Cyto Trol Control cells or cells Make a set of printouts showing pre and post sort histograms indicating sort purity and sort acquisition rate s to give to the customer 10 TURN DIAL POT PANEL MOUNT PPU TOF CARD COAX LEGEND USE 3 17 6027078 EXISTING 17 7003043 J7 Figure 3 13 1 Coax Cable Connections SLOT 5 PULSE PILE UP amp TIME OF FLIGHT PPU TOF COAX INTERCONNECT BMWDTH NORM QPSH 1 IP E SLOT 1 CH 8 OUT J4 J ce CH 1 IN CH 8 J5 STRETCH JUMPER PLS IN 2 4 Y PARAM 8 OUT 1 JUMPER 7 LOT 8 E1 TO E2 S 20 21 CH 8 IN 1 J8 CH 8 IN IN STRETCH 1 m a PLS OUT 00007 JUMPER E2 E4 J8 J1 pou 1 1 CH 8 ji SLOTS OUT OUT PARAM 8 MUX amp SCOPE Snare ADC PSH CONTROL 4237003C INSTALLATION PROCEDURES 3 SORT PERFORMANCE ESP UPGRADE ESP Equipped Units Table 3 13 3 summarizes the upgraded circuit cards on ESP units Table 3 13 3 Circuit Cards in ESP Equipped Units
128. next l l 1 1 1 1 1 l l l l 1 LED on indicates discriminator is active for this channel CARD HAS 4 OF THESE CHANNELS blE cheese M 1 Quad PSH Card BACKPLANE INTERFACE AND CONTROL CIRCUITRY P1 Backplane Connector 7003071A ADC and PSH Control Card The ADC and PSH Control card Figures 2 13 3 and 2 13 4 contains an Analog Calculator circuit that can calculate the ratio of any two inputs The voltage that represents this ratio is routed out of the card on J17 and is jumpered back into J9 where it is treated as any other incoming parameter PN 4237003C 2 13 5 2 INSTRUMENT DESCRIPTION SIGNAL SELECTION AND DIGITIZATION Figure 2 13 3 Block Diagram ADC and PSH Control Card PEAK SENSE AND DISCSAT Input to ADC card indicates HOLD CHANNEL at least one PSH channel has detected a pulse PULSE e IN PSH m OPEN ene CLOSE Control and Timing STRETCHED PULSE OPEN Circuits 1 o PSH CLOSE CHANNEL 2 CLOSE STRETCHED PULSE EE o PSH m OPE CHANNELZ3 CLOSE STRETCHED PULSE L CHANNEL 4 CLOSE STRETCHED PULSE
129. of video memory to display The sorting cursors are added to the image The processed camera image is read out from the video memory reconverted to an analog video signal and routed to the Dual CRT Control card Dual CRT Control Card The camera video signal passes through this card only if requested by the CPU To do this select DISPLAY CAMERA on the Cytometer Control Screen If you do not select DISPLAY CAMERA then the camera video ends here and the card outputs the alternate video display small histograms or service displays The Dual CRT Control card generates the menus and routes them to the right CRT for display Digiscope Card When you turn the display select knob to the camera position this card switches the camera image or alternate Dual CRT Control output In the other position the digiscope generated image is displayed Pulse Video Display The Pulse Video Display consists of the Mux and Scope card This card selects the upper and lower trace signals Digiscope card This card digitizes the selected pulses and stores them in onboard memory Logic and timing circuits use stored pulse data to provide a scan or raster compatible video signal The card generates the grid image that serves as a background for the pulses The card outputs the pulse image if the Display Select knob is in the scope position The video signal is routed to the left CRT KIR3 card and membrane panel The KIR3 card surrounds the front o
130. on the two primary power cords into the sockets at the rear of the Cytometer Note All cables that connect to the computer are routed through the side of the computer pedestal as shown in Figure 3 4 1 Figure 3 4 1 Electronic Cable Connections M POWER PRINTER B SWITCH HP LASERJET II POWER SWITCH MONITOR ad ge MOUSE TO PEDESTAL PWR COMPUTER COVER REMOVED FOR CLARITY 5 Connect the mouse keyboard and monitor to the computer as shown in Figure 3 4 2 7003061A a Connect the mouse to the COMI port on the computer b Connect the keyboard to the keyboard jack on the computer 42370036 3 4 1 INSTALLATION PROCEDURES CONNECTING ASSEMBLIES c Connect the color monitor to the Video Card connection on the back of the computer Figure 3 4 2 Connections to Workstation CPU WWW WWW r L el o COVER REMOVED A mp Q LINE VOLTAGE SELECT dest KEYBOARD TO TO MONITOR MOUSE PEDESTAL PWR CAMERA UT LT VIDEo our SERIAL COMM DATA LISTER OUT ES SERVICE e 00 e e 12467 50 7003020 6 Connect the parallel data cable a Connect one end of the cable to the DATA LISTER OUT connector on the Cytometer as shown in Figure 3 4 2 b Feed
131. pin on the bottom of target to line up with desired target hole and lock down the pin with a setscrew Install the highest target in the space between the beam translator and UV mirror on the optical rail and ensure the target is square to the table top Rotate the translator window so the UV beam is aligned with the top hole in the alignment target Remove the raised UV laser target PN 4237003C INSTALLATION PROCEDURES 3 BEAM TRANSLATOR UPGRADE 26 Use the adjustable UV laser mirror on the optical rail look at both traces on the Digiscope Screen and adjust the UV beam using the 90 degree UV mirror for the same time frame as the HeNe caused signal Between the peak of the UV and HeNe and the Argon caused signals the time separation should be 40 microseconds for the 100 pm sort sense tip at 12 psi e 60 microseconds for the 76 sort sense tip at 12psi or e 7 microseconds for the 76 um jet in air tip Note The Argon laser signal should be on the far left of the Digiscope screen Test this by increasing and decreasing the UV laser power and observing the far right peak that follows Block the HeNe laser and raise lower the Argon laser power and observe the far left signal that follows 27 Determine correct alignment by ensuring the Argon laser exits near the center of the beam shaper and the UV laser exits the beam shaper slightly below the Argon Place a business card several different places in the beam s path to verify th
132. pressure gauge does not exceed 30 psi and that the system vacuum gauge indicates at least 10 in Hg Adjust the regulator to ensure 30 psi on the gauge during operation 3 8 1 INSTALLATION PROCEDURES HOUSE AIR INSTALLATION 3 8 2 PN 4237003 INSTALLATION PROCEDURES 3 GATED AMPLIFIER UPGRADE 3 9 GATED AMPLIFIER UPGRADE Purpose Use this procedure to install the Gated Amp kit in the Elite or Elite Analyzer Instruments ordered with a Gated Amp configuration are shipped with it already installed The Gated Amp GA consists of two main sections a Switchable Amp section and a Gated Amp section The new amplifiers allow high speed low speed switching while the Peak Signal Boards allow the acquisition of FALS and PEAK signals in addition to enhancing the control of the Discriminators SAT function The following are prerequisites to installing the Gated Amp kit e Software revision 3 0 or above External Memory Upgrade in the Cytometer Multibus card cage Note If this upgrade is needed see Heading 4 13 for instructions The Gated Amp Kit is wired and packed in an anti static container Figure 3 9 1 The container is specifically made for ease of installation Cards are pre wired within the container and are ready to slide into the top card rack one at a time interconnected to each other The container is packed in a brown cardboard box The height of the cardboard box is approximately the same as the bottom lip of the
133. pressure pinch valve at the far left fitting on the valve bracket 4 Sequentially adjust three pots by a slight amount until the set pressure read pressure and gauge pressure all agree 0 05 psi a Enter the Sample SET pressure 14 psi and adjust R51 to match gauge with set b Adjust R34 to match read with SET c Enter sample pressure 5 psi and adjust R59 to match read with SET gt Repeat a through c as needed 42370036 SERVICE AND REPAIR PROCEDURES PNEUMATICS SYSTEM 4 5 Connect external gauge to sheath pressure line at the sheath pressure pinch valve center fitting 6 Sequentially adjust three pots slightly until the set pressure read pressure and gauge pressure all agree 0 05 psi Allow time for pressures to stabilize and make a series of small adjustments to approach the final values a Set Sheath Pressure to 14 0 psi b Adjust R75 to match the read and gauge pressures Note that both pressures change as the pot is turned the object is to make them agree with each other not with the Sheath SET pressure Set Sheath Pressure to 5 psi and adjust R76 to match the read and gauge pressures d Set Sheath Pressure to 14 psi and adjust R81 to match gauge with SET e Repeat steps b through d until no further adjustments are needed 7 Ifyou experience difficulty with any of these procedures repeat the adjustments in step 1 If you continue to experience difficulty with these procedures do Heading
134. regulator then the regulator is defective 8 Once sample pressure is present you can check the sample pressure display circuit a Measure voltage at TP2 Voltage should be 8 V 0 01 V b Ifvoltage at TP2 is not correct adjust R51 as needed If 8 V cannot be achieved the card is defective c Measure voltage at TP7 Voltage should be proportional to sample pressure of 8 V 0 01 V If there is no output the sensor on the card is defective PN 4237003 7 4 3 TROUBLESHOOTING TROUBLESHOOTING PRESSURE CONTROL PROBLEMS 7 4 4 4237003C TROUBLESHOOTING TROUBLESHOOTING THE SORT SUBSYSTEM 7 5 TROUBLESHOOTING THE SORT SUBSYSTEM Purpose Do this procedure to troubleshoot sort problems Use Figure 7 5 1 and Table 7 5 1 as guides Figure 7 5 1 Troubleshooting Sort Problems SORTING TROUBLESHOOTING APPLICATION Cells Beads Verify appropriate use and setting of SETUP OPERATION and ADJUSTMENT Use Procedures Verify correct e frequency Mechanical INSTRUMENT Hardware or Software Control Verify adjustment and condition of Fluidics Verify functions e stable pressures Electronics Verify functions CD waveshape level 2 e drive e flow cell stage and frequency discriminators phase e no leaks no regulator e sort
135. shaper 15x60 mm 6857111 Beam shaper 15x80 mm 6 6859007 Deflection body holder new style 6858222 Deflection body holder old style 7 6704311 Deflection body Autoclone Sorting Option 6704754 Deflection body standard 8 6859315 Pinhole snout 3X 9 6705402 Scatter sensor R2 6705182 Peak scatter sensor 10 6857400 Prism holder 11 3210004 Sample tubing 0 010 i d x 0 036 o d PN 4237003C 8 2 19 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 20 Flow Cell Area Elite Analyzer See Table 8 2 20 7003106 Table 8 2 20 Elite Analyzer Flow Cell Area Detail of Parts See Figure 8 2 20 Item Part Number Description Item Part Number Description 1 1019539 Interlock bracket 9 6856191 Flow cell tip 2 6857133 Shutter 10 6857066 Pinhole snout 3 2851558 Machine screw 25 20 x 1 00 11 1016487 Sample tubing 7 0 4 6858019 Beam shaper 15x80 mm 12 2851836 Machine screw 25 20 x 0 62 6857111 Beam shaper 15x80 mm 2687 6856826 Beam shaper 8 80 mm 5 1019009 Flow cell bracket 13 3814172 X Y axis positioning stage 6 3210004 Pinch tubing 0 010 i d 0 036 14 1020903 Tube holder bracket 7 1010933 Nut clamp 15 3814173 X Y Z axis positioning stage 8 6705597 Scatter sensor 16 N A Beam stage 8 2 20 PN 4237003C PARTS LISTS 8 ILLUSTRATED PARTS Figure 8 2 21 Deflection Body and Holder See Table 8 2 21 7003107A
136. stage are mounted Although the front of pickup lens protrudes through this plate the lens assembly mounting flange will not pass through the plate opening The following procedure must be followed to ensure correct reinstallation of the optical plate Tools Supplies Needed Allen wrench 5 32 in long or T handle recommended Allen wrench 7 64 in long or T handle recommended 3 15 1 INSTALLATION PROCEDURES COLLECTION OPTICS UPGRADE 3 15 2 Removal Procedure If materials are available it is highly recommended that a sensitivity test be performed before and after collection lens replacement As a minimum collect CV data with fluorospheres Note the HV and gain If cells or Cyto Trol Control cells are available run samples stained with red and green dyes before and after collection lens replacement l Doasensitivity test a Collect CV data or Cyto Trol control cells are available run the sample stained with the red and green dyes e Ifcells or Cyto Trol control cells are not available run fluorospheres b Record the HV and gain c Continue the upgrade procedure e If doing this procedure as instructed in step 10 of the Replacement Procedure go to step 11 of that procedure e lfdoing this procedure for the first time go to step 2 Allow the Argon laser to warm up for at least 30 minutes Remove the FALS detector and install alignment target at this location Remove pneumatically oper
137. subsystem see Laser Control Subsystem under Heading 2 5 gt Video subsystem see Video Subsystem Cytometer under Heading 2 5 gt Control Pathways subsystem see Cytometer Control Subsystem under Heading 2 5 e Acquisition subsystem see Heading 2 6 ACQUISITION SUBSYSTEM Sort subsystem see Heading 2 8 SORT SUBSYSTEM e Pneumatics subsystem see Heading 2 9 PNEUMATICS SUBSYSTEM e Electronics subsystem see Heading 2 4 ELECTRONICS SUBSYSTEM Physical Specifications Refer to Chapter 2 of the Reference manual Power Requirements Refer to Chapter 2 of the Reference manual Reagents and QC Materials Refer to Chapter 1 of the Reference manual Software The Elite operates with two software programs the Elite software and the Cytometer software For more information on the software refer to the Software manual Fluorescence and Light Scatter Measurement Overview The system measures the fluorescence and laser light scattered from cells or from other microscopic particles passing through a laser beam Samples must be single cell or particle suspensions the system measures up to 10 000 cells per second The intensity of fluorescence gives the affinity of the sample for certain dyes or of the inherent fluorescence of the sample particles There are two types of scattered light forward and side Forward light scatter correlates with particle size Side light scatter correlates with the granularity or complexity of a cel
138. the number of steps to move each stepper to position the tray correctly This sequence must be successfully completed before the Autoclone Sorting Option can operate Calibration also needs to be performed if the movement of the mechanism is interfered with a limit switch has been activated or the movement speed has been changed Autoclone Sorting Option Mechanism Adjustment The speed of movement the step rate of the motors must be adjusted to optimize operation of the system The X Y speed is adjusted to provide smooth movement of the mechanism from left to right and in and out Rotational speed of the tray is controlled by the ROT speed The rotation is adjusted so it is smooth and slow enough that liquid in the wells will not be spilled Calibration must be performed after each speed change to ensure the mechanism does not overtravel and strike a limit switch Refer to Chapter 6 of the Options Manual for instructions on how to operate the Autoclone Sorting Option For installation instructions refer to Heading 3 10 AUTOCLONE SORTING OPTION INSTALLATION PN 4237003C INSTRUMENT DESCRIPTION 2 OPTICAL COMPONENTS 2 3 OPTICAL COMPONENTS Figure 2 3 1 illustrates the various optical components of the Elite Figure 2 3 1 Optical Components Top View with Cover Removed 488 MIRROR BEAM REDUCER EXPANDER FLUORESCENCE HENE LASER ARGON LASER BEAM WATER COOLED COLLECTION OPTICS LASER 2 LASER 1 EXPANDER ARGON LASER wem
139. the engineering schematics you need for troubleshooting the Elite flow cytometer Electronic pdf files of these schematics are available on a separate CD ROM in the Service Resource Kit SRK and in a Lotus Notes database The schematics in the SRK are the latest revisions available at the time the SRK is released For copies of schematics released between revisions of the SRK check the Lotus Notes database It will always have the most current revisions Note Depending on the configurations of this instrument in the field more than one revision of a schematic can be valid If you want to include schematics in the printed version of this manual make printouts of the electronic files and insert them at the end of this chapter Most Frequently Used Schematics Gated Amp Interconnect PN 6319353 Pneumatic Hydraulic Layout PN 6320201 Interconnect Diagram Laser Interlock PN 6320234 Pneumatic Hydraulic Layout Analyzer PN 6321140 Additional Schematics for All Systems Acquisition Card Cage Lower Bitmap And Sort Decision PN 6319569 Data Acquisition Backplane PN 6319561 Data Lister Out R PN 6319520 Interface and Scaler R PN 6319535 MUX amp Scope Interface PN 6319541 Peak ADC PSH Card ESP PN 6320856 PRISM amp Sort Window Test PN 6319570 Pulse Gen amp Clock PN 6320194 Pulse Pileup Det TOF 2 ESPt PN 6321443 Quad Peak Sense amp Hold PN 6319559 Sort Delay R 3 ESP t PN 6321511 Sort Oscillator R 2 ESP PN 6321026 Sort O
140. the logic conditions sort criteria and makes a final decision to sort gt Waits for the cell to be in the last attached droplet gt Tests the cell for coincidence gt Deflects the droplet containing the desired cell Circuit Cards Figure Figure 2 8 2 shows the Sort subsystem circuit card functions 4237003 2 8 1 INSTRUMENT DESCRIPTION SORT SUBSYSTEM 8705004 1 A 0005 sasind ups w104 ayeound 9949 19 09 9904 s jdoup jo jequinu P eseudisnipy SHEM Avid Reap eg 0149019 JISE 9121999 HOS 0 aziuo1upu S syuana ajqeuonsenb IUH 0 110 uos 1 10 3191 10 eag 310 pue Dog 2511 sdeung suoifau swopuiM 1sureDe sanje 1S8 dwy 9 Jase yno sassed aed uaum 10 W014 pue 59204 sane 5910049919 Uonisinboy lt udiouig je1s 49 964245 weas asing uos Maan Waysksgng 110 9 z 4 PN 4237003C 2 8 2 2 INSTRUMENT DESCRIPTION SORT SUBSYSTEM VIGO0E00 301 1 U3MOd div 0309
141. the original non delayed pulse 3 divisions Change the delay to 40 microseconds and adjust R99 on the same card as above Change the delay to 60 microseconds and adjust R103 as above Change the delay to 7 microseconds 3 9 10 PN 4237003C INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE Figure 3 9 4 Gated Amplifier Assignment and Adjustment Potentiometer Locations PN 4237003C x 6560 9 3 Hz 3OV4u3INI HOSNAS 911015 101S3HVdS 9101 28 909 e 0181 d L Y HILIMS AQ LWIS 9107 69050 ek 0181 G u HOLIMS XNW LYIS ed 1078 25 2 9250 9 gis 0151 nee S Y HOLIMS INS Ide 6101 60009 888 8 o oklekloxzesz 0181 E 03199 Xnv lV2S 1016 M co o o zzo 0670 9 5 2 oz Ros SENTIS non A70 09 07 08 QVO 01016 z e s Sl el 2 el e z 0181 L AY130 0900 61018 5 si all e 9 2 SE amp 6 e Sie E d m Z 8 1016 r P co o o 0690 9 2 Oz N Y LI Z 09 05 02 GYND 1018 5 5l g 8 e 9059
142. their implementation of Acquisition Start Delay ASD also called discriminator extension ASD allows the operator to control the length of time that the acquisition system samples the signals from each cell The length of time must be properly set otherwise problems could result The time must be long enough to allows all signals sufficient time to reach their maximum values yet short enough to avoid noise or signal contamination from other particles When determining and setting an optimum time consider Hardware factors gt Flow cell type gt Laser beam size gt Amplifier operating mode fast or slow e Protocol requirements gt Types of signals being acquired peak integral or log gt Which signal is used for the discriminator channel Changes in any of the above factors may require you to change the ASD Note The type of ADC used determines the settings available to the operator 2 13 8 PN 4237003C INSTRUMENT DESCRIPTION 2 SIGNAL SELECTION AND DIGITIZATION Peak ADC Card The Peak ADC card supports ASD operation The Cytometer gives the user the option of 2 5 5 10 or 15 microseconds of ASD Note Full Pulse Width FPW is not an option with this card See Figure 2 13 5 Figure 2 13 5 ASD Settings Diagram e oe 7003039A DISCSAT EXT DISCSAT DISCSAT EXT DISCSAT 4 MICROSECONDS 15 MICROSECONDS 4 MICROSECONDS 2 5 MICROSECONDS Poor ASD setting time is too Poor ASD setting time is too short
143. then PARAMB 1 m Most problems that occur with the instrument are fluidic related and some of the fluidic problems result in the same symptoms noted in this table Always check fluidics before doing an alignment procedure Table 7 1 6 Electronics General Troubleshooting Information Problem Probable Cause Corrective Action System locks up Power problems Power down the system wait 30 seconds then power up Table 7 1 7 Software General Troubleshooting Information Problem Probable Cause Corrective Action Touch Screen locks up Power problems Power down the system wait 30 seconds then power up Workstation locks up Power problems Press Ccti alt Delete to reboot the computer OR Turn computer off wait 30 seconds then turn computer on Table 7 1 8 Power Supplies General Troubleshooting Information Problem Probable Cause Corrective Action No sort deflection Problem with 3000 V deflection supply Check and or replace First alligator clip is not attached to the insertion rod Attach alligator clip properly No stream undulation or deflection Problem with 36 V and 90 V power supplies Measure power supply and replace if defective See Heading 4 4 POWER SUPPLIES MEASUREMENT AND ADJUSTMENT Camera stops moving camera cannot be adjusted incorrect readings on the Control Screen Problem with 1
144. tips New flow cell tips and collection lens assemblies are available as a purchased upgrade for existing Elite systems Systems manufactured after September 1993 incorporate these items as standard These are readily identified by noting that the new collection lens is noticeably smaller The new flow cells have a smaller lens and no mirror The flow cell tip types are e Flow cell tip 100 p e Flow cell tip 76 p Flow cell tip Biosense The new flow cell tips can be used without replacement of the collection optics assembly This does require removal of the collection lens focus stop to allow sufficient focusing adjustment Tools Supplies Needed One of the following flow cell tips gt Flow cell tip 100 p PN 6859300 0 gt Flow cell tip 76 PN 6859313 1 Flow cell tip 3x PN 6859341 7 Pinhole snout 6859315 8 Snout Upgrade kit PN 6913251 1 0 Power meter Flow Cell Replacement Procedure Note If system does not have the 3x collection lens assembly remove the collection lens focus stop on the side of the PMT housing Caution the customer not to strike flow cell with collection lens 1 Remove the existing flow cell tip 2 Install the new flow cell tip Snout Upgrade Procedure Purpose Removal of the fluorescent pickup lens assembly first requires removal of the vertical optical plate This is the plate on which the FALS detector the beam shaping stage and the flow cell adjustment
145. to E2 Connect your DVM to TP11 Connect tee your pressure gauge to the sheath regulator output At the Cytometer Control screen set Sheath Pressure to 12 psi DVM should read 8 00 V a lfnot do steps 1 through 3 of Pneumatic Interface Card Adjustment under Heading 4 5 b Ifyou cannot obtain the indicated voltages the circuit card is defective or the card is not receiving all the necessary power supply voltages Check J10 for 15 15 24 45 V c Ifyou still cannot obtain the correct reading the Pneumatic Interface card is defective or the card is not being correctly set by the CPU Replace the card or inspect replace the ribbon cable to Sensor I O card If you obtained the correct reading in step 4 the regulator is being set for 12 psi Measure the air pressure at the regulator output If it is 12 psi the pressure is acceptable If the pressure is not correct adjust the regulator a Adjust the Range pot on the regulator until you read 12 psi At the Cytometer Control screen set Sheath Pressure to 6 psi The voltage at TP11 should change to 4 00 V If not there is an electronic problem that must be corrected before proceeding See step 4 c Adjust the Zero pot so the gauge reads 6 psi d Repeat steps a through c until the pressure is correct at both levels If the correct pressures cannot be achieved and the correct pressure 30 psi is available to the regulator the regulator is defective Once the r
146. top card rack Figure 3 9 1 Anti Static Container of Gated Amp Kit LEGEND 15 14 13 12 1110 9 8 SLOT CARD NAME Gated Amp Control R 3 Dual FL SW R 3 PMT SUB SW R2 3 PMT SUB SW R1 PMT Gated Amp Quad 20 40 60 Microsecond Delay 2 Quad 7 Microsecond Delay 2 Quad 7 Microsecond Delay 1 10 Quad 20 40 60 Microsecond Delay 1 11 Scat Aux Gated Amp 12 3 PMT SUB SW R 13 Peak Scatter Mux SW R 14 Scat CV SW R1 15 Mux and Scope Interface gt 7003016 Tools Supplies Needed C type extender card PN 6702299 One of the following kits gt Gated Amp Service Field Upgrade Kit for units before SN U07122 PN 6912827 gt Gated Amp Service Field Upgrade Kit for units after SN U07122 PN 6912798 Alignment Tool PN 5402071 Oscilloscope PN 4237003C 3 9 1 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE 3 9 2 Procedure 1 tr ouo 00 D 10 11 12 13 Examine the packing list for the kit and ensure all parts are present Notify shipping of any discrepancy Verify operation of all signals and unit function before beginning kit installation Turn OFF power at Cytometer and Workstation Remove the tabletop Ensure cables from PMTs and FALS sensor are labeled Remove the existing Scatter Sensor R Pre Amp card and replace with the new Peak Scatter Sensor card connecting P105 to J35 and P83 to J36 Reinstall the fo
147. uofuy yids LXnv se 929 Duiubissy od up xmv se paubisse 99 01 S 2LINd SMOJE uonouny SIUL N9ISSV ANY LXnv 9919 dwy pajer 9 6 4 3 9 15 PN 4237003C INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE 3 9 16 18 19 At the Cytometer s Scope screen a Put PMTI INT on the bottom and PMT2 INT on the top b Adjust PMT2 HV and gain to get PMT2 to three divisions Put PMT3 INT on the top trace and repeat as above then do 5 if installed Put the mirror into the dichroic slot under PMT4 and adjust its voltage and gain as above Go back to the scope screen and select a parameter a for the top trace AUX 1 Adjustment See Table 3 9 3 for additional information on adjustments 1 At the Cytometer s Scope screen a Set Trace 1 to PMTI b Set Trace 2 to PMT2 2 Atthe Cytometers Gated Amp Assign screen select a AUX1 SOURCE for PMT2 b AU SOURCE for PMT2 3 At the PARM A box ensure AUX1 INT is selected and at the PARM B box ensure AUX2 INT is selected Adjust the preamp gain so that pulse is three divisions tall 4 Select AUX1 for UPBM Adjust R112 on the Quad 7 Microsecond Delay card slot 9 so that the AUXI pulse is the same amplitude as the non delayed original PARM A pulse three divisions 5 Goto the Gated Amp Control Screen and change the delay to 20 microseconds Note The followin
148. voltage is stored until Input Buffer Ensuring that the entire pulse the FET is turned on and the capacitor is discharged Gain 1 is captured by the PSH circuit This readies the card for another pulse This transistor is switched on Stretched pulse after pulse is captured to block to ADC and PSH any noise input after the pulse TP2 Control Card WM v rh ES 02468 48 87 pi Gs This comparator detects if and when D the input signal exceeds the 4 and Scope Card Vv for X Y display FET Signals to ADC and PSH input CLOSE signal from ADC closes PSH input once capacitor is charged DISCSAT signal to ADC Reference voltage 10 000V This channel can only send a DISCSAT informs ADC that a valid from ADC and PSH Control Card to the ADC if this gate is enabled The OFF signal has been detected discriminator value turns this gate off and ensures this channel cannot trigger conversion Latch The discriminator Digital to analog converter 3 gt 45V Produces a voltage from A value channel number kel is stored here gt lt discriminator discriminator level l 1 1 1 1 1 1 1 l l l l 1 1 1 1 G Control Card ADC M OPEN signal from ADC wv resets capacitor for
149. which is controlled by the CPU card The CPU card commands the Dual Laser Control card to turn on the laser PN 4237003C ALL KEYSWITCH INTERLOCK AND LAMP CONNECTIONS ARE ON THE P2 CONNECTOR INSTRUMENT DESCRIPTION CONTROL AND DISPLAY SUBSYSTEMS 2 Figure 2 5 1 Laser Control Subsystem FOR ADDITIONAL LASER ON LASER INTERLOCK LAMP CONNECTIONS e gt 1 LAGER KEYSWITCH INTERLOCK OVERRIDE KEYSWITCH m B LL LASER COVER INTERLOCK SWITCHES LASER CONTROL CARD From air cooled laser socket ARGON ON 5 V INTERFACE HENE LASER LOGIC P1 MULTIBUS CONNECTOR SOLID STATE RELAY AC from Main Xformer SOLID STATE 10006 RELAY lt RELAY EN gt A C 9 Power 4 RELAY Supply LASER AC In INTERLOCK CONNECTOR PN 4237003C AIR COOLED ARGON LASER POWER SUPPLY l ARGON LASER 7003063A 2 5 3 INSTRUMENT DESCRIPTION CONTROL AND DISPLAY SUBSYSTEMS Video Subsystem Cytometer This subsystem produces the displays on the two CRT screens indicated in Table 2 5 1 Table 2 5 1 CRT Display CRT Display Source Right Control screens menus Cytometer CPU car
150. 0 tip and 3000 V power supply 3 Oneata time remove each card listed below from the system and examine it for revision level as listed in Table 3 13 1 ATTENTION If the revision level is not correct then return the card to the authorized repair center Do not make changes in the field PN 4237003 3 13 1 INSTALLATION PROCEDURES SORT PERFORMANCE ESP UPGRADE Table 3 13 1 Circuit Card Revision Level Circuit Card Part Number Minimum Revision Level Task Peak Scatter Mux 6705369 C Ensure C98 is 3 pF Ensure C99 is 10 pF Ensure R67 is 100 ohms 3 PMT Sub Amp 6705246 Ensure capacitor locations C136 C134 C132 are not present Ensure C114 C85 C58 are 5 pF Ensure C133 C135 C137 are 24 pF Scatter Sensor 6705182 Ensure U1 is a OP61A PMT Gated Amp 6704008 Ensure capacitor locations C147 193 59 C24 are not present Ensure diode locations CR19 CR14 CR11 are not present Ensure U18 is AD746 Ensure R205 is 1000 ohm Ensure R40 is 4 99 kohm Ensure R36 is 10 kohm Ensure that near U41 U30 U19 U10 a diode was added to the card near each of these op amps The diode s anode is electrically connected to pin 8 All diode cathodes are connected to ground Scat Aux Gated Amp 6704009 Ensure capacitor locations C98 C67 C41 C20 are not present Ensure diode locations CR19 CR10 CR15 CR6 are not present Ensure U1 is AD746 Ensure R143 is 1000 ohm Ens
151. 0 16 6028086 Cable assembly fluid detector 8 2 18 15 6028086 Fluid detector SN4 cable behind sample station 8 2 18 15 6028092 Pneumatic PCB to sheath transducer cable W45 6028122 Bimorph cable 6028144 Camera video cable ground mixer 6028210 Back flow linkage 8 2 10 1 6028211 Serial 9 25 pin IBM adapter 1 ft long 6028218 Cable assembly with interlock Argon laser 8 2 34 5 6028256 Interlock cable HeCd laser 8 2 38 4 6028266 Flex circuit cable Autoclone Sorting Option 14 in 8 2 23 2 3 6028268 Cable main Autoclone Sorting Option 8 2 25 1 6028278 SCSI host ribbon cable 6028302 Power module control 25 pin external 6028304 Cytometer 12 6028371 Fiber optics glass 6028374 Analyzer card W52 8 2 10 2 6028884 Data lister out EMC 6856953 Harness W12 8 2 10 6 6858355 Connector panel Elite Analyzer 8 2 10 15 PN 4237003C 8 PARTS LISTS MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY Table 8 1 4 Cards Part Number Description Figure Item 2016127 PC Bus monochrome 25 in long 2016378 Memory Expansion 2 MB 32 bit 8 2 28 7 2016413 Video Elite VGA SW setting 2016424 Controller Intel disk floppy 8 2 28 6 2016526 Computer 486 33 ISA 8 MB RAM 2016545 1522A Adapter SCSI Host Tahiti 2306042 Easy Artist crystal oscillator clock 6702608 Sort Transistor 8 2 9 22 8 2 12 17 6702664 Serial 1 0 Card 8 2 9 2 8 2 11 3 6703761 DT Interface 8 2 9 3 8 2 11 4 6703772 Digiscope 8
152. 0 FLT README DOC HRVGA SYS INSTALL FLT INSTALL EXE MOUSESYS SYS HELP TEX ELITE EXE MSMOUSE SYS TOPICS TXT CONTROL HEX LASERJET SYS CYTOMETE BAT IBMEGA SYS HELP TEX IBMGPR SYS TOPICS TEX DESKJET SYS DUMP EXE HPPJ90 SYS INSTALL FLT HPPJ180 5YS CHAR4S FLT IHM6X8 FTN CHAR90 FLT IHM12X16 FNT IHM24X24 FNT HETA SYS 7003015A EPSONX SYS PN 4237003C FILE PATHS LOCATION OF SOFTWARE FILES B 1 2 PN 4237003C ABBREVIATIONS The following list is a composite of the abbreviations acronyms and reference designators used in this manual When the same abbreviation or reference designator is used for more than one word or type of component all meanings relevant to this manual are included separated by semicolons SYMBOL wavelength of laser A change in difference between Q ohm gt greater than less than subtract negative plus plus or minus equal to u micron micro amperes uf microfarad uL microliter um micrometer microsecond degrees Celsius F degrees Fahrenheit A A ampere A D analog to digital ac alternating current adpt adapter adj adjustment amp amplifier Ar Argon assy assembly auto automatic aux auxiliary avg average BK blocking BMWDTH beam width PN 4237003C BNC connector bayonet Neil Concelman connector BP band pass Btu British thermal units C C centigrade capacitor C
153. 0 Microsecond Delay card from the extender and install into slot 10 15 Put the Quad 20 40 60 Microsecond Delay card slot 7 onto the extender 16 Turn on the instrument 17 After Elite software is loaded select GATED AMP protocol 18 Press F9 19 At the Cytometers Options screen set the Align Led to pulsed 100 and 2kHz 20 At the Cytometers Scope screen select PMT1 INT for the lower and PMT2 INT for the upper trace Note You should see two traces each with a three division high deflection body 2 Press 10 PMT 2 Adjustment See Table 3 9 3 and Figure 3 9 4 for additional information on adjustments 1 At the Cytometer s Gated Amp Control screen select 7 microseconds delay Verify that PMT2 INT pulse upper trace is 3 divisions tall Note the amplitude of upper trace PMT2 INT At the Gated Amp Assign screen select PMT2 as UPBM Adjust R105 on the Quad 7 Microsecond Delay card slot 8 so the pulse is three divisions tall At the Cytometer s Gated Amp Control screen set the delay to 20 microseconds Adjust R95 on the Quad 20 40 60 Microsecond Delay card so the pulse is three divisions tall Change the delay to 40 microseconds and adjust R99 so the pulse is three divisions tall Change the delay to 60 microseconds and adjust R103 as above for three divisions tall Set the delay to 7 microseconds PMT 3 Adjustment See Table 3 9 3 and Figure 3 9 4 for additional information on adjustments
154. 1 At the Scope screen select INT as the top trace Note the amplitude of PMT3 INT pulse three division tall trace At the Gated Amp Assign screen select PMT3 UPBM Adjust R107 on the slot8 card so that the pulse is three divisions tall 3 9 17 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE 3 9 18 Change the delay to 20 microseconds and adjust R96 on the slot 7 card as above 5 Change the delay to 40 microseconds and adjust R100 on the slot 7 card as above 6 Change the delay to 60 microseconds and adjust R104 on the slot 7 card to get the amplitude as above PMT 4 and 5 Adjustment See Table 3 9 3 and Figure 3 9 4 for additional information on adjustments 1 Repeat the 3 Adjustment procedure for 4 and 5 if installed When finished turn power off remove the extender and insert card into slot 7 Remove the mirror from the PMT 3 slot and put it in the PMT 4 or PMT 5 slot as needed Offset adjustments for PMT 4 and PMT 5 as shown in in Table 3 9 4 and Figure 3 9 4 Table 3 9 4 Offset Adjustments Non Delay 7 Microsecond Delay 60 Microsecond PMT Slot Adjustment Slot Adjustment Slot Delay Adjustment FALS 11 R135 9 R106 10 R107 PMT1 11 R136 9 R111 10 R108 AUX1 11 R137 9 R115 10 R109 AUX2 11 R138 9 R119 10 R110 PMT2 6 R197 8 R106 7 R107 PMT3 6 R198 8 R111 7 R108 PMT4 6 R199 8 R115 7 R109 5 6 R200 8 R119 7 R110 Applies only
155. 1 Sort Kit Installation 3 13 3 ESP Equipped Units 3 13 5 ENTERPRISE LASER UPGRADE 3 14 1 Purpose 3 14 1 Tools Supplies Needed 3 14 1 Important Operating Notes 3 14 2 Installation 3 14 2 COLLECTION OPTICS UPGRADE 3 15 1 Purpose 3 15 1 Tools Supplies Needed 3 15 1 Flow Cell Replacement Procedure 3 15 1 Snout Upgrade Procedure 3 15 1 Purpose 3 15 1 Tools Supplies Needed 3 15 1 Removal Procedure 3 15 2 Replacement Procedure 3 15 3 PIEZOELECTRIC TRANSDUCER INSTALLATION 3 16 1 Purpose 3 16 1 Tools Supplies Needed 3 16 1 Installation 3 16 1 Initial Flow Cell Alignment 3 16 2 xi xii Alf 3 18 3 19 3 20 3 21 3 22 WATCHDOG TIMING 3 17 1 Purpose 3 17 1 Tools Supplies Needed 3 17 1 Procedure 3 17 1 SOFTWARE VERSION 4 0 INSTALLATION 3 18 1 Purpose 3 18 1 Tools Supplies Needed 3 18 1 Procedure 3 18 2 SOFTWARE VERSION 4 01 INSTALLATION 3 19 1 Purpose 3 19 1 Tools Supplies Needed 3 19 1 Installation Procedure 3 19 2 Waste Bottle Modification for Software Version 4 01 3 19 2 COLOR PRINTER UPGRADE 3 20 1 Purpose 3 20 1 Tools Supplies Needed 3 20 1 Printer Setup 3 20 1 Software Installation 3 20 1 Editing CGI CFG File DOS 6 0 and Higher 3 20 2 Editing CGI CFG File Lower Than DOS 6 0 3 20 4 Printer Test 3 20 6 PC MODEL PENTIUM 133 MHz UPGRADE 3 21 1 Purpose 3 21 1 Tools Supplies Needed 3 21 2 Procedure 3 21 2 ELITE 256 SORT
156. 14 X X X X X 0 0 Open the jumper settings for the 0 0 0 0 0 0 External Memory card SW6 0 0 0 0 0 0 SW7 X X X 0 0 X SW9 0 0 0 0 E19 E20 open E25 E26 jumpered External Memory X1 jumpered X6 a 3 pin jumper jumpered to the 12 V position Camera Interface E19 E20 E21 E22 E37 E39 E38 E42 E41 E45 E44 E46 Dual CRT Control E11 E12 E13 E14 Digiscope E69 70 E31 39 E25 17 SW ALL ON Dual Laser Control 4 7 E25 E26 28 29 E31 E32 E34 E35 E37 E38 E40 E41 Non Gated Amp Card Cage Table A 2 2 Non Gated Amp Card Cage Jumpers Card Jumpers Gated Amp Control R2 E4 E5 E7 E8 3PMT Sub Amp SW R E9 E11 Dual FL Amp SW R E8 E9 E11 E12 Scat CV Amp SW R E9 E11 E5 E7 Peak Scatter Mux SW R E1 E2 E3 E4 E5 E7 Sensor Interface 2 E5 E6 E8 E9 11 12 E13 E14 E16 E17 E19 E20 E22 E23 A 2 2 4237003 QUICK REFERENCE INFORMATION A JUMPERS AND SWITCH SETTINGS Gated Amp Card Cage Table A 2 3 Gated Amp Card Cage Jumpers Card Jumpers Gated Amp Control R3 None Dual FL Amp SW R E8 E9 E11 E12 3 PMT Sub Amp SW R2 4 9 11 3PMT Sub Amp SW R1 E9 E11 PMT Gated Amp None Quad 20 40 60 Microsecond Delay 2 E1 E2 E3 E4 E10 E11 In Quad 7 Microsecond Delay 2 E10 E15 In 1 Quad 7 Microsecond Delay E10 E11 In Quad 20 40 60 Microsecond Delay 1 E1 E2 E3 E4 E9 E10 In Scat Aux Gated Amp None 3PMT Sub Amp SW R E1 E2 E5 E6 E9 E11 Peak Scatter Mux SW R E1 E2 E3
157. 1600C Color Printer cyan Cartridge ink HP 1600C Color Printer yellow Guide installation Elite color printer Cable parallel printer PN 4237003C 8 1 21 8 PARTS LISTS MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY Table 8 1 18 Printers Continued Part Number Description Figure Item 6417268 Driver software HP 1600C Color Printer 6914968 Kit HP 1600C Color Printer 6914969 Kit Elite color printer Table 8 1 19 Regulators Part Number Description Figure Item 6232490 Low Bleed Regulator sample regulator 8 2 5 1 Used with Pneumatic Interface Card PN 6705504 6232491 Low Bleed Regulator sheath regulator 8 2 5 2 Used with Pneumatic Interface Card PN 6705504 Table 8 1 20 Software Part Number Description Figure Item 6141379 Flownet Profile 3 5 in 6414357 Profile 2 26 3 5 in 6414520 DOS 4 01 5 25 in 6414852 OPTune 3 5 in 6414853 OPTune 5 25 in 6414976 Elite version 4 00 disk 1 6414977 Elite version 4 00 disk 72 6912816 Elite version 4 0 with PrintQue 6913114 Elite version 4 01 6915099 Elite version 4 5 Table 8 1 21 Tools Part Number Description Figure Item 4717730 Potentiometer dual CRT trigger adjustable 8 2 7 9 7232508 Enterprise laser hex wrench 8 1 22 PN 4237003C MASTER PARTS LIST IN NUMERICAL ORDER CATEGORY Table 8 1 22 Tubing PARTS LISTS Part Number Description Figu
158. 16468 Cartridge data removable disk 21 4 MB 2016500 Ribbon printer color Stare 2016501 Cartridge ribbon printer black Star NX 2480 2016526 Card Adapter SCSI host 2016527 Printer dot matrix Panasonic 2016545 Kit adapter SCSI host 2016551 Cartridge disk removable 90 MB 2016556 Cartridge printer font Microsofte HP Laserjet 2016557 Printer letter tray Laserjet Il P Plus 2016558 Cartridge printer Deskjet 500 2016560 Cartridge toner printer Laserjet P Plus 2016568 Drive disk optical rewritable 512 650 MB 2016569 Drive disk optical 650 MB write 1x Worm 512 2016570 Drive disk optical 1 GB write 1x Worm 1024 2016576 Cartridge toner printer Laserjet IV 2016577 Switch data manual A B 2016592 Keyboard 101 key compact 2016613 Computer 486 DX2 66 MHz 16 MB RAM ISA 2016615 Cartridge toner printer Laserjet 4P 2016616 Printer Laserjet 4 2016651 Monitor 17 in 1024x1024 8 2 2 5 2016652 Drive disk cleaning kit 3 5 in 2016670 Computer Pentium 90 MHz 16 MB RAM 540 MB hard disk drive 8 2 2 2 2016690 Card Memory PCMCIA 512 K 2121537 Connector T BNC UG 274B 2429044 Keyboard overlay function strip long 2430458 Keyboard template 2XL label 2906841 Terminator BNC male 50 Ohm 0 5 W 4004048 Battery 3 0V lithium 4004070 Battery 3 6V NiCad 6027510 Cable printer parallel 6027937 Cable data transfer Cytologic 9 pin 6027939 Cable data transfer Cytologic 25 p
159. 19 8 1 20 8 1 21 8 1 22 8 1 23 8 2 1 8 2 2 8 2 3 8 2 4 8 2 5 8 2 6 8 2 7 8 2 8 8 2 9 8 2 10 8 2 11 8 2 12 8 2 13 8 2 14 8 2 15 8 2 16 8 2 17 8 2 18 8 2 19 8 2 20 8 2 21 8 2 22 8 2 23 8 2 24 Cables 8 1 2 Cards 8 1 4 Covers Doors 8 1 7 Cytometer 8 1 7 Filters 8 1 10 Flow Cell Area 8 1 11 Fuses 8 1 12 Hardware 8 1 12 Kits 8 1 15 Lamps 8 1 16 Lasers 8 1 16 Miscellaneous 8 1 18 Optics 8 1 19 Pneumatics 8 1 19 Power Supplies 8 1 21 Printers 8 1 21 Regulators 8 1 22 Software 8 1 22 Tools 8 1 22 Tubing 8 1 23 Workstation 8 1 23 Elite Cytometer Right Panel Removed Detail of Parts See Figure 8 2 1 8 2 1 Workstation Detail of Parts See Figure 8 2 2 8 2 2 Sheath and Waste Compartment Detail of Parts See Figure 8 2 3 8 2 3 Pneumatics Cabinet Detail of Parts See Figure 8 2 4 8 2 4 Pneumatics Drawer Detail of Parts See Figure 8 2 5 8 2 5 Pneumatics Panel Rear View Detail of Parts See Figure 8 2 6 8 2 6 Dual CRT and Connector Panel Detail of Parts See Figure 8 2 6 8 2 7 Elite Card Configuration Detail of Parts See Figure 8 2 8 8 2 8 Elite ESP Card Configuration Detail of Parts See Figure 8 2 9 8 2 9 Elite and Elite Analyzer Interconnections Detail of Parts See Figure 8 2 10 8 2 10 Multibus Card Cage All Configurations Detail of Parts See Figure 8 2 11 8 2 11 Elite ESP Data Acquisition Card Cage Detail of Parts See Figure 8 2 12 8 2 12 Cytometer I
160. 2 PMT Gated Amp Card 2 12 4 Function 2 12 4 Inputs 2 12 4 20 40 60 Microsecond Delay Card 2 12 6 7 Microsecond Delay Card 2 12 6 SIGNAL SELECTION AND DIGITIZATION 2 13 1 Overview 2 13 1 Signal Flow Description 2 13 2 Signal Source 2 13 2 Mux and Scope Card 2 13 2 Quad Peak Sense and Hold PSH Cards 2 2 13 3 ADC and PSH Control Card 2 13 5 ADC and PSH Timing 2 13 8 Peak ADC Card 2 13 9 ADC and PSH Control Card 2 13 11 Lister Out Card 2 13 12 Lister A B Card 2 13 14 ILLUSTRATIONS 2 1 1 2 1 2 2 1 3 2 2 1 2 3 1 2 3 2 2 3 3 2 3 4 2 3 5 2 3 6 2 3 1 2 3 8 2 3 9 2 3 10 Elite Components 2 1 1 Sensor Area 2 1 3 Sample Stage 2 1 3 Autoclone Sorting Option Mechanism 2 2 1 Optical Components Top View with Cover Removed 2 3 1 Beam Shaper Options 2 3 6 Alignment Mirror 2 3 8 Beam Expander Reducer 2 3 9 Sort Sense Flow Cell Original 2 3 10 100 n Sort Sense Flow Cell Cut away View 2 3 10 76 um High Velocity Quartz Flow Cell 2 3 11 Optical Filters Location 2 3 12 Forward Scatter Detector 2 3 13 Forward Scatter Obscuration Templates 2 3 14 PN 4237003C 2 2 4 Block Diagram System Electronics 2 4 3 2 5 Laser Control Subsystem 2 5 3 2 5 2 Block Diagram Video Subsystem 2 5 4 2 5 3 Control Pathways 2 5 7 2 6 1 Subtraction Circuit Example 2 6 2 2 6 2 Log Amp Response 2 6 3 2 6 3 Log Amp Response Log Lin Axes 2 6 3 2 6 4 Circuit Operation 2 6
161. 2 1 Purpose 5 2 1 Tools Supplies Needed 5 2 1 Before You Begin 5 2 1 Procedure 5 2 1 6 SCHEMATICS 6 1 1 6 SCHEMATICS REQUIRED 6 1 1 7 TROUBLESHOOTING 7 1 1 7 1 ISOLATING THE PROBLEM AREA 7 1 1 Purpose 7 1 1 Tools Supplies Needed 7 1 1 Procedure 7 1 1 General Troubleshooting 7 1 2 XV T 3 74 P TO TROUBLESHOOTING THE ACQUISITION SUBSYSTEM 7 2 1 Purpose 7 2 1 Tools Supplies Needed 7 2 1 Procedure 7 2 1 Acquisition Troubleshooting Methods 7 2 3 Subtraction Color Compensation 7 2 3 PMT 3 PMT 2 Compensation 7 2 3 Log Amplifiers 7 2 4 TROUBLESHOOTING FLUIDIC PROBLEMS 7 3 1 Purpose 7 3 1 Tools Supplies Needed 7 3 2 Air Leaks 7 3 2 Sheath Air Leak Testing 7 3 2 Liquid Leak Testing 7 3 2 Sheath Liquid Leak Testing 7 3 2 TROUBLESHOOTING PRESSURE CONTROL PROBLEMS 7 4 1 Purpose 7 4 1 Tools Supplies Needed 7 4 1 Procedure 7 4 1 Sheath Pressure Problems 7 4 1 Sample Pressure Problems 7 4 3 TROUBLESHOOTING THE SORT SUBSYSTEM 7 5 1 Purpose 7 5 1 Tools Supplies Needed 7 5 1 Sort Current Leakage Test 7 5 1 RESETTING CYTOMETER SOFTWARE LOCKUPS 7 6 1 Purpose 7 6 1 Tools Supplies Needed 7 6 1 Reset the Cytometer CPU 7 6 1 Clear the CPU s Memory on the Cytometer 7 6 1 8 PARTS LISTS 8 1 1 8 1 8 2 MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY 8 1 1 ILLUSTRATED PARTS 8 2 1 A QUICK REFERENCE INFORMATION A 1 1
162. 2 7 1 Refraction of Light 7003174A 24 1 2 INSTRUMENT DESCRIPTION OPTICAL ELEMENTS 2 1 2 Snell s Law Refraction at the interface between two optical media with indices of refraction n1 and n2 appears as the following according to Snell s Law nl sin 01 n2 sin 02 where the angle is measured normal to the surface In most materials n is a function of the light s wavelength This property is known as dispersion and it allows a prism to separate colors However this is a problem for lenses since lights of different colors refract differently and therefore focuses differently Lenses The Elite uses two types of lenses positive and negative Positive Lens A positive lens has two convex spherical surfaces and e Reduces a laser beam to cellular size since a laser is a source of parallel rays e Forms an image Figure 2 7 2 shows the positive lens focus All parallel rays of light that enter the lens refract so they all converge on a point known as the focal point The lens works as well in reverse The lens refracts those rays emitted from the focal point in varying directions into a parallel beam of light The Elite s collection lens applies this principle to gather the fluorescent light leaving the cell into a parallel beam directed toward the filters and the PMTs Figure 2 7 2 Positive Lens Focus FOCAL POINT lt FOCAL gt 7003175 LENGTH Image Formation The camera lens projects and magnifi
163. 2 FL1 OI The other 3 signal coax to 3 Sub Amp Sw 1 FL3 J3 Remove coax from FL2 in J2 on 3 PMT Sub Amp SW R 1 card Disconnect BNC from PMT 3 and remove cable from instrument Connect two short BNC bullet coax cables to the PMT 4 BNC T connector Label the bullet ends as PMT 4 and connect a One PMT 4 signal coax to 3 PMT Sub Amp SW RI FL2 in J2 b Second PMT 4 signal coax to 3 PMT Sub Amp SW R2 FL in J2 Connect PMT 5 signal coax to 3 PMT Sub Amp SW R2 FL3 in J3 Connect 3 Int coax cable to Sub Amp SW RI FL3 Int Out 08 Connect coax to 3 PMT Sub Amp SW R1 FL3 Peak Out 06 Connect 3 log coax to PMT Sub Amp SW RI FL3 Log Out J7 Connect PMT 4 Int signal to Dual FL SW R FL2 Int Out J7 Connect PMT 4 Pk signal coax to Dual FL SW R FL2 Pk Out J6 Connect PMT 4 Log signal coax Dual FL SW R FL2 Log Out J8 Label one end of a 27 in bullet bullet coax PMT 5 Int Label the other end MUX J22 Label one end of a 27 in bullet bullet coax PMT 5 Log Label the other end MUX J23 Label one end of a 27 in bullet bullet coax PMT 5 Peak Label the other end MUX J24 Connect the PMT 5 Int signal coax to 3 PMT Sub Amp SW R2 FL3 Int Out J8 Connect the PMT 5 Log signal coax to 3 PMT Sub Amp SW R2 FL3 Log Out J7 Connect the 5 Peak signal coax to 3 Sub Amp SW R2 FL 3 Peak Out 6 Connect the MUX J22 end of the coax to Mux and Scope 22 Connect t
164. 2 and PMT4 Peak c Main Adjust PMT4 high voltage for PMT4 peak pulse of about 5 V See Figure 4 12 15 Obtain maximum pulse heights a Adjust the flow cell horizontal knob as needed b Adjust the flow cell vertical knob as needed c Adjust the laser beam vertical knob as needed d Adjust the fluorescence pickup lens Figure 4 12 16 for simultaneous maximum pulse heights Repeat step 3 at least once Obtain minimum pulse widths by adjusting the focus knob and collect new histograms and recheck the statistics See Figure 4 12 17 for the knob location 4 12 9 4 SERVICE AND REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE Figure 4 12 15 PMT4 High Voltage Setting PMT4 PEAK 5 vors 2 B 7003202A Figure 4 12 16 Fluorescence Pickup Lens FLUORESCENCE PICKUP LENS 0401545710 7003193A 4 12 10 PN 4237003C SERVICE AND REPAIR PROCEDURES 4 OPTICAL ALIGNMENT PROCEDURE Figure 4 12 17 Focus Knob Location EXTERNAL FOCUS ADJUSTMENT KNOB 7003205A PMT5 System ATTENTION Risk of losing traces Follow the sequence of steps to avoid losing the traces when setting up the scope l Atthe Cytometer select a 1 and PMT2 Peak b 2 and B c Main Options gt gt Gated Amp Assign gt gt PARAM B AUX2 Peak d Source box under AUX2 and PMT5 Note 5 peak pulse now appears on the upper trace trace 2 e
165. 2 and check stream position Repeat steps 6 through 8 as needed Note You may need to readjust the tray to arm parallelity for optimal alignment Remove the tray and tray holder and select Retract on the screen Observe that the arm moves all the way back Connect the fan assembly cable to the fan on the new outside cover assembly PN 4237003C INSTALLATION PROCEDURES AUTOCLONE SORTING OPTION INSTALLATION 11 Connect the hard tubing coming through the opening on the frame to the fitting on the cylinder 12 Connect the soft tubing to the fitting on the beak 13 Install the outside cover assembly ensuring that tubing and cables are out of the way of the tray 14 Repeat steps 4 through 9 and ensure that neither the arm nor the tray assembly hits the outside cover If it does check the cover for proper installation 15 Install the Autoclone Sorting Option cover into sample collection area securing with the knob to front of the Autoclone Sorting Option arm 16 Remove the beak assembly from the old sample collection drawer and install into bezel cover assembly 17 Transfer sample collection drawer accessories from old cover to new cover 18 Install the new deflection body assembly 19 Reinstall and secure all panels and covers removed in this procedure Adjusting Autoclone Sorting Option Speed 1 Atthe Cytometer screen select Options vr Autoclone Sorting Option gt gt Control Adjust Screen 2 Adjust the Autoclon
166. 237003C 4 10 3 4 SERVICE AND REPAIR PROCEDURES ELECTRONIC ADJUSTMENTS CABLE AND JUMPER CONFIGURATIONS Table 4 10 5 Data Acquisition Card Cage Cards Jumper Settings Slot Card Jumpers 15 Sort Oscillator R2 None 16 Sort Output R None 4 10 4 Analyzer HV DAC Control Card l Jumper EI E5 E2 E6 E3 E7 E4 E8 2 Install Analyzer HV DAC Control card in the far left slot of the top Bertan card cage behind the Data Acquisition card cage 3 Connect blue ribbon cable P85 to first top Analyzer HV DAC connector PI Note Pin 1 must be on top 4 funit has a Gated Amp 5PMT option a Ensure that the second HV DAC Control card has the following jumpers E5 E9 E6 E10 E7 E11 E8 E12 b Install the second HV DAC Control card in the bottom Bertan card cage in the far left slot c Connect blue ribbon cable P86 to the second lower Analyzer HV DAC connector P1 Note Pin 1 must be on top PC Model 486 Jumper the 486 pc first slot on the left Lister AT card E71 E72 Cable Connections Procedure Ensure that the cables are connected as shown in Table 4 10 6 and the connectors as shown in Table 4 10 7 Table 4 10 6 Cable Connections From To Blue Ribbon Cables Interface and Scaler J1 P70 Pin 1 on top Bitmap and Sort Decision J1 P38 Pin 1 on bottom Lister Out J1 P43 Pin 1 on bottom Mux and Scope J27 P88 Pin 1 on top Sensor Interfac
167. 25 kHz Remove the bandpass filters from under the PMTs leave the dichroic filters in place At the Workstation a Create a protocol to acquire PMTI 90 degree light scatter and PMT2 as single parameter histograms b Set the Discriminator for PMTI to 100 set all others to OFF 4 Acquire the FL test pulses and adjust gains and high voltages as needed to obtain histograms 5 Atthe Workstation Sort screen a Create a region to include the population acquired from PMT2 b Name the region Test Press to return to the Sort screen Use the mouse to select the Test region in the region list box When selected the Test region turns white 8 Move the mouse cursor to the box at the lower center of the screen and click on one of the ID numbers Enter the test name Test when prompted Note The name and sort equation appear next to the ID number selected Do not enter any equations into the SORT RIGHT or SORT LEFT boxes 9 Atthe Cytometer s Autoclone Sorting Option screen select a Plate and choose 96 well plate b Orifice Diameter and enter flow cell tip being used 100 c Well Timeout and enter 1 second d Test equation e Fill Plate and select Fill Plate again 10 Enter a count at the prompt 10 11 When prompted for Description press Enter The plate displayed on the screen fills with the ID number of the selected equation First Test 1 FL Test DRIVE to 0 2 Select Activate The Autoclone Sorting Option s
168. 2806201 Screw RPL sample drawer 6 32 x 0 38 8 2 17 8 2807038 Setscrew jet in air ring 6 32 x 0 250 8 2 16 4 2839058 Screw self locking sample drawer 4 40 x 0 18 8 2 17 6 PN 4237003C 8 PARTS LISTS MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY Table 8 1 6 Cytometer Continued Part Number Description Figure Item 2851876 Thumbscrew RPL sample drawer 6 32 x 0 25 8 2 17 7 3814172 Stage flow cell 8 2 19 2 8 2 20 14 3814173 Stage beam 8 2 19 3 8 2 20 16 3814223 Mirror 45D adaptor 8 2 29 3 8 2 36 6 3814271 Positioner ultra compact flow cell stage 8 2 31 1 8 2 32 1 4506029 Relay K1 and K2 2 pole 120 Vac 8 2 27 2 4508007 Relay K3 and K4 100 140 Vac 3 32 Vdc input 8 2 27 2 5120109 Switch drip chamber 8 2 5 3 5120178 Switch pressure 25 psi 8 2 5 6 5120196 Switch rotary dual CRT 8 2 7 10 5120199 Switch waste filter 5609034 Transformer T3 100 220 240 8 2 27 4 5609035 Transformer T1 and T2 100 220 240 8 2 27 5 6027967 Cable assembly ground 7 50 in 8 2 14 6 6028002 Cable assembly ground 14 ga 5 0 in 8 2 14 4 6211015 Valve actuator pilot silver 8 2 18 14 23 8 2 19 1 6214106 Valve check for 0 125 in tubing 8 2 6 11 6214108 Valve check 6232205 Fitting barbed hose sample drawer elbow 0 125 i d to 10 32 nylon white 6703950 Backplane HV DAC PMT 8 2 13 3 6704171 Backplane data acquisition card cage 8 2 12 18 6704753 Camera stage 8 2 1 1 6704754 Deflection
169. 2unos XNY pue 1999 0 7 aures BY JE eui 0199109 3 9115 SMOJE SIU Jguueuo xf ue se 99 0 G ZLINd uonoun 141 N9ISSV ZXNY ANY LXNY 1 payey e e e aJnBiJ 3 9 9 PN 4237003C INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE FALS Adjustment Assignment and Adjustment Potentiometer Location Note When in the delay Enable section of the Gated Amp Assignment screen Figure 3 9 3 LWBM means no delay and UPBM means delay is on See Table 3 9 3 and Figure 3 9 4 for additional information on adjustments l Atthe Gated Amp Control screen Figure 3 9 5 a Ensure FS INT is on the top trace and PMTI INT is on the bottom trace Note amplitude of FS INT on scope upper trace 3 divisions b Change the delay to 7 microseconds then go to the Gated Amp Assignment screen Figure 3 9 6 c Select FALS for the UPBM and record the amplitude of the FALS trace on Scope upper trace 2 Adjust R105 on the Quad 7 Delay 1 slot 9 so the delayed pulse has the same amplitude as the original FALS pulse 3 divisions 3 Atthe Gated Amp Control screen change the delay to 20 microseconds ATTENTION The following adjustment must be made on the component side of the Quad 20 40 60 card slot 10 4 the component side of the Quad 20 40 60 Card slot 10 adjust R95 so the delayed FALS pulse has the same amplitude as
170. 3 1 Purpose 4 13 1 Tools Supplies Needed 4 13 1 Procedure 4 13 1 ILLUSTRATIONS 4 1 1 Camera Assembly Adjustment 4 1 12 4 2 1 Waveforms 4 2 1 4 2 2 Front and Back Porch 4 2 6 4 2 3 Pulse Signals and Histograms for Sort Coincidence Verification 4 2 14 4 4 1 Power Supplies 4 4 1 4 11 1 Fittings on Valve Assembly Bracket 4 11 2 4 11 2 Connecting Clip from Flow Cell to DVM 4 11 3 4 11 3 Filament Connecting Last Attached Drop 4 11 6 4 12 1 Interlock Defeat Bar 4 12 1 4 12 2 Location of Optical Alignment Components 4 12 2 4 12 3 Laser Beam Reflected through Shutter Hole 4 12 2 4 12 4 Flow Cell Bodies Old and Newv 4 12 3 4 12 5 New Flow Cell Body 4 12 4 4 12 6 Old Flow Cell Body 4 12 4 4 12 7 Parabolic Mirror and LED 4 12 5 4 12 8 Laser Shutter Location 4 12 5 4 12 9 Laser Beam Centered on Forward Scatter Detector Mask 4 12 6 4 12 10Scope Screen Selections 4 12 6 4 12 11 Laser Beam Horizontal Adjustment Location 4 12 7 4 12 12Laser Beam Centered Across Forward Scatter Detector 4 12 7 4 12 13Flow Cell Z Axis Knob Location 4 12 8 4 12 14 PMT2 High Voltage Setting 4 12 9 4 12 15 PMT4 High Voltage Setting 4 12 10 4 12 16 Fluorescence Pickup Lens 4 12 10 PN 4237003C 4 iii 4 iv 4 12 17 Focus Knob Location 4 12 11 4 12 18PMT5 High Voltage Setting 4 12 12 4 12 19 Focus Knob Location 4 12 12 4 13 1 TABLES 4 2 1 4 3 1 4 4 1 4 5 1 4 10 1 4 10 2 4 10 3 4 10 4 4 10 5 4
171. 3 GIOAV Q3 11V343Q 390 1H31NI N3dO N3HM NOILLVIQVH H3SV1 3 8lSIANI HO QNYV 3 18lSIA aa auniuadv SIHL SI NOILVIGVH H3SV 1 HO GNV 3 181SIA 3unsodxa 1V343 90 TH31N ea1y Buisuag ui sjaqe Gulmem pL Z e PN 4237003C 3 7 14 INSTALLATION PROCEDURES 3 OPTIONAL LASER INSTALLATION Figure 3 7 15 Warning Lahels Rear of Instrument THIS LASER PRODUCT COMFORMS TO THE PROVISIONS OF 21 CFR SUBCHAPTER J SECTION 1040 10 AND 1040 11 VISIBLE AND OR INVISIBLE LASER RADIATION WHEN OPEN AVOID EYE OR SKIN EXPOSURE TO DIRECT OR SCATTERED RADIATION Manufactured DATE 19 by EPICS DIVISION OF COULTER CORP Hialeah FL 33010 7003094A 35 Verify interlock operation by interrupting the optical area safety cover a Verify that all covers are installed b Turn on the Cytometer c Power up Laser 1 Laser 2 and appropriate optional laser Result All three lasers should turn on d Lift open optical area safety cover Result All three lasers should shut down e Turn on all three lasers Result Lasers should not turn on with cover open Verify that the certification label and rear cover warning label are in place Note Ensure that the external blower draws cooling air through the laser head when the laser is turned ON 36 Replace covers before resuming operation
172. 31 32 33 At the Gated Amp Assignment screen Figure 3 9 6 a Put PMTI to LWBM and block the PMT 1 signal by putting one of the blanks supplied with the filter kit into the bandpass slot under it b Adjust R136 slot 11 to minimize any pulse on the upper trace during or after the window c Select PMT1 UPBM and adjust R111 slot 9 to minimize any pulse present on the upper trace during or after the window Note You may increase delay and open window up to the right of the signal At the Gated Amp Control screen Figure 3 9 5 a Setthe delay to 60 microseconds and the clock to 5 b Adjust R108 slot 10 to minimize any pulse present on the upper trace c Disable the trigger gate d Set the delay to 7 microseconds and the clock to 0 5 At the Gated Amp Assign screen a Set AUXI and AUX2 source to PMT2 b Set PARM A to PMT2 LOG PARM B to PMT2 LOG At the Scope screen put PARM A on the top trace At the Gated Amp Control screen enable GATED AMP At the Gated Amp Assign screen a Change AUXI to the LWBM and block PMT2 signal by moving the blank into the bandpass slot b Adjust R137 slot 11 to minimize any pulse present on the upper trace c Change AUXI to UPBM d Adjust R115 slot 9 to minimize any pulse present on the top trace At the Gated Amp Control screen a Change the delay to 60 microseconds and the clock to 5 b Move the gate and windows out to right c Adjust R109 slot 10 to minimize any pul
173. 4 sort performance temperature affect of 7 5 3 Sort subsystem block diagram 2 8 3 block diagram circuit card functions 2 8 3 circuit cards functions of 2 8 1 function 2 8 1 Multibus CPU card function 2 8 1 overview 2 8 1 sort cycle 2 8 1 sort waveforms See waveforms sort sort wells electrostatic buildup of See Autoclone Sorting Option sort window See window sort SP short pass filter function 2 3 12 stream See sort function stretched pulse defined 2 13 4 subtraction function See color compensation switchable amplifiers definition of SW and SWITCH designators 2 6 5 description 2 6 5 integrator circuit illustration of 2 6 6 pulse width variation of 2 6 5 switcher power supply adjustments and measurements 4 4 2 A 3 2 switches 16 INDEX drip chamber switch function 2 9 1 limit Autoclone Sorting Option function of 2 2 3 mechanical Autoclone Sorting Option function of 2 2 2 2 2 3 overtravel Autoclone Sorting Option function of 2 2 2 pressure switch adjustment See pressure switch adjustment system pressure function of 2 9 1 system pressure in Compressor module 2 9 1 waste filter function of 2 9 1 sync function verifying synchronicity of sort pulses with crystal drive 4 2 4 system test procedures 3 6 1 system electronics troubleshooting 7 1 6 system pressure valve VL1 function of 2 9 2 T Table 3 21 1 Technical Specs 3 21 1 temperature affect on sorting performance
174. 4 5096 and ensure Trace 2 decreases by 5096 Put back to 096 Replace 575 BP with 640 DL filter under PMT 4 At the Scope Screen a b Select PMT 4 Peak for Trace 1 Select PMT 3 Peak for Trace 2 Place the 550 DL filter under PMT 3 Check for three division tall pulses for 3 4 and 5 Adjust HV if necessary At the FL Compensation Screen a C d Select PMT 4 minus PMT 3 for 5096 and ensure Trace 1 decreases by 5096 Put back to 096 Select PMT 3 minus PMT 4 for 5096 and ensure Trace 2 decreases by 5096 Put back to 096 Put PMT 5 Peak on Trace 2 Select PMT 5 minus PMT 4 for 5096 and ensure Trace 2 decreases 5096 Put back to 096 Put all light filters back in proper positions in slot Turn off Align LED 3 11 5 INSTALLATION PROCEDURES 5TH PMT KIT WITH SWITCHABLE AMPS INSTALLATION PROCEDURE 3 11 6 PN 4237003 INSTALLATION PROCEDURES BEAM TRANSLATOR UPGRADE 3 12 BEAM TRANSLATOR UPGRADE Beam translators are available as upgrade parts the translators are not part of the modification kit for the HeCd laser The customer can order the kit using PN 6912985 4 The kit contains Three Beam Translators one for each laser UV light either HeCd or Argon UV laser PN 6858244 0 488nm light Argon laser PN 6858245 8 633nm light HeNe laser PN 6858243 1 Three micro optical rails for mounting PN 3814206 2 Two machine screws for each rail that mounts to the optical bench PN 2851763 7 The
175. 4 6703761 Data Taker Interface card 5 7000681 External Memory card Note Use only with the Computer IBC 86C card PN 6706571 61 6704660 Camera Interface card 7 6704089 Dual CRT Control card 8 6703772 Digiscope card 9 6704463 Dual Laser Control card 10 2603033 Fan blower 24 Vdc 73 CFM On units manufactured after 10 30 96 t Elite Analyzer does not contain this component PN 4237003C 8 2 11 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 12 Data Acquisition Card Cage Elite ESP See Table 8 2 12 7003142A Table 8 2 12 Elite ESP Data Acquisition Card Cage Detail of Parts See Figure 8 2 12 Item Part Number Description Item Part Number Description 1 6704906 Pulse Generator and Clock card 10 6704142 Data Lister Cut card 2 N A Spare 11 6704185 Prism and Sort Window Test card 3 N A Spare 12 6704186 Bitmap and Sort Decision card 6706232 Bitmap and Sort Decision R2 card 4 N A Spare 13 6704147 Interface and Scaler card 5 6705954 Pulse Pileup Det TOF card 14 6704198 Sort Delay card 6 6704164 Mux and Scope Interface card 15 6704188 Sort Oscillator card 7 6704169 Quad PSH card 2 16 6704201 Sort Output card 8 6704169 Quad PSH card 1 17 6702608 Sort Transistor card 9 6704139 ADC and PSH control card 18 6704171 Data acquisition backplane 8 2 12 PN 4237003C PARTS LISTS 8 ILLUSTRATED PARTS Figure 8 2 13 Cytometer Inside Rear Vi
176. 5 8 Multibus CPU card communication pathways to Pneumatic Interface card 2 9 4 function 2 5 1 Multibus subsystem function 2 5 1 Mux and Scope card block diagram 2 13 3 42370036 cables functions of 2 13 2 function 2 5 5 2 13 2 jumpers 3 9 2 A 2 1 A 2 3 jumpers illustration of 3 9 3 signal flow summary of 2 6 9 Mux and Scope Interface card jumpers 4 10 3 A 2 3 ND neutral density filter function 2 3 13 negative lens description 2 7 3 neutral density ND filter function 2 3 13 noise isolating card s input from 2 13 4 Norgren See pressure regulators Notice of Information Update form purpose 1 1 1 use 1 1 1 0 operating clearances required for Cytometer 3 2 2 operational tests See VIP optical components illustration of 2 3 1 optical drive connections of 3 4 4 optical elements definition of light 2 7 1 flow cell tips high sensitivity description 2 7 8 image formation description 2 7 2 lenses 2 7 2 light sources 2 7 8 negative lens description 2 7 3 overview 2 7 1 parallel plate function 2 7 7 positive lens description 2 7 2 refraction of light description 2 7 1 optical filters BK blocking function 2 3 12 BP band pass function 2 3 12 DL dichroic long pass function 2 3 12 DS dichroic short pass function 2 3 13 function of overall 2 3 11 LP long pass function 2 3 12 ND neutral density function 2 3 13 replacement when to do 5 2 2 SP s
177. 5 V 5A power supply Measure power supply and replace if defective See Heading 4 4 POWER SUPPLIES MEASUREMENT AND ADJUSTMENT PMT high voltages cannot be adjusted Problem with 15 V 3 4A power supply Measure power supply and replace if defective See Heading 4 4 POWER SUPPLIES MEASUREMENT AND ADJUSTMENT 42370036 TROUBLESHOOTING v ISOLATING THE PROBLEM AREA Table 7 1 8 Power Supplies General Troubleshooting Information Continued Problem Probable Cause Corrective Action No signals Problem with 15 V Measure power supply and replace if defective See Heading 4 4 0 25A power supply POWER SUPPLIES MEASUREMENT AND ADJUSTMENT Unable to make screen Problem with 5 V Measure power supply and replace if defective See Heading 4 4 selections computer locks power supply POWER SUPPLIES MEASUREMENT AND ADJUSTMENT up No image on CRTs wavy Problem with 12 V Measure power supply and replace if defective See Heading 4 4 lines on displays or CPU power supply POWER SUPPLIES MEASUREMENT AND ADJUSTMENT problems Solenoids do not fire Problem with 24 V Measure power supply and replace if defective See Heading 4 4 Power Supply POWER SUPPLIES MEASUREMENT AND ADJUSTMENT PN 4237003 7 1 7 TROUBLESHOOTING ISOLATING THE PROBLEM AREA 7 1 8 4237003C TROUBLESHOOTING TROUBLESHOOTING THE ACQUISITION SUBSYSTEM 7 7 2 TROUBLESHOOTING THE ACQUISITION SUBSYS
178. 600C Color Printer PN 6417268 9 gt Installation Guide for Elite Color Printer PN 4237252 8 HP 1600C Ink Cartridges kit PN 6914967 7 which includes gt Magenta ink cartridge PN 2016691 6 gt Black ink cartridge PN 2016692 4 gt Cyan ink cartridge PN 2016693 2 gt Yellow ink cartridge PN 2016694 1 Printer Setup 1 Unpack the Printer and verify all items are included 2 Setup the Printer according to the instructions supplied with the printer 3 Install the parallel printer cables from the rear of the Printer to the parallel port at the rear of the Elite Workstation Software Installation 1 Atthe Workstation keyboard press to exit to DOS 2 Insert the Driver for HP 1600C diskette into the 3 5 in drive 3 Type the 3 5 in drive letter A or B and press Enter PN 4237003 3 20 1 INSTALLATION PROCEDURES COLOR PRINTER UPGRADE 3 20 2 COPY C ELITE GSS and press Enter Type and press Enter Remove the diskette from the drive Verify that ELITE is the prompt displayed Type CD GSS and press Ente Verify the DOS version Edit the CGI CFG file for the HP 1600C Printer according to the DOS version instructions under either Editing CGI CFG File DOS 6 0 and Higher or Editing CGI CFG File Lower Than DOS 6 0 Editing CGI CFG File DOS 6 0 and Higher Type EDIT CGI CFG and press Enter Locate the printer driver without a REM statement before it This should
179. 7 Socket modified 8 2 26 2 6028266 Cable flex circuit 14 in 8 2 23 2 3 6028268 Cable main Autoclone Sorting Option 8 2 25 1 6232037 Cylinder air single acting 8 2 24 7 6704311 Deflection body 8 2 19 7 8 2 26 3 6704313 Autoclone Sorting Option mechanism 8 2 25 3 6705191 Card Autoclone Sorting Option 8 2 25 2 6705328 Card Interconnect 8 2 23 1 6705329 Card Position Detection B 2 23 8 6856635 Bezel cover 8 2 26 1 6858187 Tray support sub assembly 8 2 23 10 6858880 Beak Autoclone Sorting Option 8 2 24 4 6912869 Kit Autoclone Sorting Option 8 2 23 PN 4237003 8 1 1 PARTS LISTS MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY Table 8 1 2 Beam Shapers Part Number Description Figure Item 6856826 Standard 6 5 um ht 151 um width 8 mm front focal length 80 mm 8 2 20 4 rear focal length sub assembly for package PN 6604660 6858427 Optional 8 um ht 151 um width 10 mm front focal length 80 mm rear focal length PN 6604895 6856696 Standard 12 um ht 51 um width 15 mm front focal length 60 mm 8 2 19 5 rear focal length PN 6604657 6857111 Optional 12 um ht 70 um width 15 mm front focal length 80 mm 8 2 19 5 rear focal length PN 6604656 8 2 20 4 6858019 Standard 12 um ht 151 um width 15 mm front focal length 80 mm 8 2 20 rear focal length PN 6604658 6857269 Standard 6 um ht 112 um width 15 mm front focal length 80 mm rear focal length PN 6604659 6857630 Standard
180. 7003 2 2 1 INSTRUMENT DESCRIPTION AUTOCLONE SORTING OPTION 2 2 2 Mechanical Assembly Description Motors The assembly contains three stepper motors two move the collection tray from left to right and from front to back and the third motor rotates the collection tray clockwise CW or counterclockwise CCW The motors rotate a discrete number of degrees whenever the current is applied to their multiple phase windings The tray rotation motor is inside the arm on top of the assembly and directly rotates the collection tray holder Rotation of the motor shaft is monitored by two optical sensors that detect when the motor shaft is in the normal position ready for a tray to be loaded or in 180 degree rotated position A third optical sensor detects the presence of the cover assembly installed by the operator when the Autoclone Sorting Option is retracted and not in use Tray Rotation Arm The tray rotation arm is mounted on a translation stage that moves from left to right when driven by a stepper motor This movement is monitored by optical sensors that move with the stage Holes drilled in a stationary metal strip trigger one of the optical sensors at each extreme of the stages normal travel If the stage overtravels in either direction a mechanical switch closes alerting the electronics and the operator that an error has occurred The left right stage is mounted to the forward backward movement stage This stage is also moved by
181. 71 Laser mounting brackets and support plate 7 3814233 Optical table 8 2851762 Machine screws metric M6x25 mm 8 2 34 PN 4237003C Figure 8 2 35 HeNe Laser Mounting Hardware See Table 8 2 35 7003012A OPTICAL TABLE VIEWED FROM REAR PARTS LISTS ILLUSTRATED PARTS Table 8 2 35 HeNe Laser Mounting Hardware Detail of Parts See Figure 8 2 35 Item Part Number Description Item Part Number Description 1 6857424 Duct plenum 9 6912985 Beam reducer expander 2 1019658 Air duct bracket 10 6857611 Beam splitter Argon 488 3 2523549 Spring 0 25 o d 11 6857612 Beam splitter HeNe 633 4 6857515 HeNe Laser 12 6856681 Argon mounting upright 5 6857112 HeNe mounting feet 13 6857516 Argon laser 6 6028016 Laser control W49 cable 14 2814004 Screw 25 20 x 0 50 7 6011002 Wire tie 6 7 x 0 14 nylon 15 6857347 Laser intake duct 8 N A Mounting plate PN 4237003C 8 2 35 8 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 36 Laser Placement Top View See Table 8 2 36 1 2 TOP VIEW Di 7003128 Table 8 2 36 Laser Placement View Detail of Parts See Figure 8 2 36 Item Part Number Description 1 6912749 Red HeNe laser laser 2 2 6912750 Argon laser laser 1 3 6912792 Innova 305 Coherent laser 4 3814286 325 mirror reflects 325 to 365 nm 5 6856298 Beam reducer expander part of kit PN 6912946 6 3814223 488 m
182. 8 2 35 8 2 35 Laser Placement Top View See Table 8 2 36 8 2 36 Cytometer Inside Right Rear View See Table 8 2 37 8 2 37 Omnichrome HeCd Laser See Table 8 2 38 8 2 38 File Paths B 1 1 PN 4237003C PN 42370036 TABLES 2 2 1 2 3 1 2 3 2 2 3 3 2 5 1 2 6 1 2 6 2 2 7 1 2 8 1 2 8 2 2 9 1 3 1 1 3 1 2 3 1 3 3 1 4 3 7 1 3 9 1 3 9 2 3 9 3 3 9 4 3 10 1 3 11 1 3 13 1 3 13 2 3 13 3 3 21 1 4 2 1 4 3 1 4 4 1 4 5 1 4 10 1 4 10 2 4 10 3 4 10 4 4 10 5 4 10 6 4 10 7 7 1 1 7 1 2 11 3 7 1 4 7 1 5 7 1 6 7 1 7 7 1 8 7 3 1 7 5 1 8 1 1 CONTENTS Autoclone Sorting Option Screen Marker Definitions 2 2 3 Laser Summary 2 3 2 Laser Technical Specifications 2 3 3 Working Beam Spot Dimensions 2 3 7 CRT Display 2 5 4 Dye Emissions 2 6 6 Configuration Summary 2 6 8 Typical Applications of Beam Shapers 2 7 6 Coincident Condition Responses 2 8 12 Power Supply Functions 2 8 19 Solenoid Valves Manifold 2 9 2 Power Requirements 3 1 2 Water Specifications for Coherent 305 Laser 3 1 2 Environmental Requirements 3 1 3 Required Operating Clearance 3 1 4 Possible Laser Configurations 3 7 1 Gated Amp Jumper Configuration 3 9 4 Coax Cable Interconnections 3 9 5 PMT Adjustments 3 9 12 Offset Adjustments 3 9 18 Spade Lug Connections 3 10 5 Card and Cable Connections 3 11 3 Circuit Card Revision Level 3 13 2 Additional Information on Cards 3 13 2 Circuit Cards in ESP Equipped
183. 80 mm Elite Analyzer 8 2 20 4 6857066 Pinhole assembly Elite Analyzer 8 2 20 10 6857111 Beam shaper 15 x 80 mm Elite Analyzer 8 2 20 4 6857133 Shutter single acting Elite Analyzer 8 2 19 4 8 2 20 2 6857388 Valve pinch double acting 8 2 19 1 6857400 Prism holder 8 2 19 10 PN 4237003C 8 1 11 8 PARTS LISTS MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY Table 8 1 8 Flow Cell Area Continued Part Number Description Figure Item 6858019 Beam shaper 15 x 80 mm 8 2 20 4 6858222 Deflection body holder 3 000 V old 8 2 19 6 6859007 Deflection body holder new 8 2 19 6 6859300 Flow cell tip 100 u 3x 8 2 31 5 6859313 Flow cell tip 76 u 3x 8 2 31 5 6859314 Flow cell tip Profile 3x 6859315 Pinhole Snout 3x 8 2 19 8 6859397 Flow cell tip ESP Sort Sense 140 8 2 31 5 Table 8 1 9 Fuses Part Number Description Figure Item 5102002 250 0 13A NB fuse 5102007 250 V 3A 5102013 250 3A slo blo 5102021 250 V 2A slo blo 5102026 250 V 0 25A slo blo Table 8 1 10 Hardware Part Number Description Figure Item 1005697 Fitting barbed metal 0 062 i d 8 2 6 7 1007908 Pin threaded Autoclone Sorting Option 8 2 24 6 1010932 Nut upper insertion rod gland 8 2 31 3 8 2 32 2 1010933 Clamp nut Elite Analyzer 8 2 20 7 1015427 Fitting restrictor tubing bottom 0 020 i d 1016827 Plate support upright rear HeCd laser 8 2 33 7 1017261 Plate front mountin
184. 9 Quick connect connector orange 2 6232092 Quick connect connector white 12 1020382 Waste bracket 3 6232451 Quick connect body white 13 6232304 Quick connect body orange 4 2851668 Paddle latch 14 6232489 Gas filter waste 5 6857374 Sheath bottle assembly 15 6232450 Quick connect connector blue 6 6028142 Sheath service cable assembly 16 1020382 Sheath bracket 7 6857373 Waste bottle assembly 17 6232309 Quick connect body blue 8 6856678 Water trap assembly 18 6232473 Water filter 0 2 micron 7260050 Clear bowl assembly 7260051 Filter element kit 9 6232452 Quick connect connector yellow 19 6216286 Muffler breather 10 6232303 Quick connect body yellow 20 2523515 Hidden hinge PN 4237003C 8 2 3 8 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 4 Pneumatics Cabinet See Table 8 2 4 7003086A Table 8 2 4 Pneumatics Cabinet Detail of Parts See Figure 8 2 4 Item Part Number Description Item Part Number Description 1 6856859 Table optical laser mount 8 6232092 Quick connect body white 2 6857872 Rinse tank assembly 9 6028142 Sheath service cable 3 6704552 Pneumatic Interface card 10 1020380 Rinse bracket 4 2603012 Fan blower 11 6202814 Cable assembly 5 6856889 Pneumatics door 12 6232451 Quick connect connector white 6 6856677 Compressor module 13 6232450 Quick connect connector blue 7 6232309 Quick connect body blue 14 6856539 Pneumatics drawer
185. ADE PC MODEL PENTIUM 133 MHz UPGRADE 3 21 4 IRQ4 IRQ5 IRQ7 IRQ9 IRQ10 IRQ11 IRQ12 IRQ14 IRQ15 Chipset Setup CPU TO PCI BURST WRITE CPU TO PCI POSTED WRIT BUS PARKING AT CPU PCI MASTER BURST LENGTH VGA PALETTE SNOOPING PCI MASTER LATENCY TIMER NON CACHABLE BLOCK 1 NON CACHABLE BLOCK 2 Power Management Disabled Peripheral Setup N BOARD FLOPPY CONTROLLER CLKS P P INFORMATI VE INFORMATI VE Y NO SELECTION SA SA PCI PNP SA SA SA SA Z Dp D E 5 A e o onore g N IO N BOARD PRIMARY SECONDARY IDE CI IDE CARD PRESENT IN PORT 1 PORT 1 FIFO PORT 2 PORT 2 FIFO PARALLEL PORT SABLED Y NO SELECTION PCI PNP FOR OPTICAL DRIVE PN 4237003C INSTALLATION PROCEDURES ELITE 256 SORT UPGRADE 3 22 ELITE 256 SORT UPGRADE Purpose Use this procedure to install and set up the Elite 256 SORT Upgrade kit Tools Supplies Needed Elite 256 Sort Upgrade kit PN 6915082 which contains gt Bitmap and Sort Decision R2 card PN 6706232 gt Elite version 4 5 Software PN 6915099 gt Elite ESP Reference Manual PN 4235904 Procedure Install the New Bitmap and Sort Deci
186. ADIATION AVOID EYE OR SKIN EXPOSURE TO DIRECT OR SCATTERED RADIATION ARGON ION LASER 12 WATTS MAN CW PRACTICAL LIMIT CLASS IV LASER PRODUCT VISIBLE AND INVISIBLE LASER RADIATION IS EMITTED AVOID EXPOSURE FROM THIS APERTURE 7003160A Figure 1 2 7 Laser Warning Labels Coherent Spectrum Laser VISIBLE LASER LIGHT AVOID EXPOSURE TO DIRECTOR SCATTERED LIGHT ARGON AND KRYPTON MIXED GAS LASER 8 WATTS MAX CW MULTILINE OUTPUT PRACTICAL LIMT CLASS IV LASER PRODUCT 7003161A PN 4237003C INTRODUCTION SAFETY PRECAUTIONS Figure 1 2 8 Laser Warning Labels Uniphase Red HeNe Laser CLOSE lt OPEN BEAM SHUTTER LASER RADIATION AVOID DIRECT EXPOSURE TO BEAM LASER RADIATION IS EMITTRD FROM THIS APERTURE HELIUM NEON 20MW CLASS LASER PRODUCT 7003158A Figure 1 2 9 Laser Warning Labels Melles Griot Green HeNe Laser AVOID EXPOSURE Laser light is emitted from this aperture OPEN SHUTTER gt CLOSE LASER SHUTTER mw THIS LASER PRODUCT 05 LHP 991 COMPLIES WITH DHHS CDRH MFG 0 NOVEMBER 1994 RADIATION S N 27478T PERFORMANCE DH ME ICD AMI SUE CHAPTER RLSBAD CALIFORNIA 92008 LASER LIGHT AVOID DIRECT EXPOSURE TO BEAM HELIUM NEON LASER 32 mW MAX OUTPUT AT 632 8 nm CLASS LASER PRODUCT S PAT 4 311 969 7003096A 4237003 1 2 7 SAFETY PRECAUTIONS INTRODUCTION
187. AL calibration cbl cable cfm cubic foot per minute Ch channel CK choke CL cylinder cm centimeter Coin coincidence Coin Abrt coincidence abort CPU central processing unit CRT cathode ray tube CV coefficient of variation D D degree diameter of laser beam D A digital to analog DAC digital to analog converter dc direct current DCN document control number det detector DI dichroic diam diameter DL dichroic long pass DMS data management system DS dichroic short pass DUAL FL Dual FL SW R card DVM digital voltmeter ABBREVIATIONS 1 ABBREVIATIONS E ea each EPROM erasable programmable read only memory ESD electrostatic discharge F f focal length of lens F Fahrenheit function fuse FALS forward angle light scatter FF feed through fitting FIFO first in first out FL filter fluorescence FPW full pulse width FRU field replaceable unit FS forward scatter ft foot FT T fitting FU fitting union FY Y fitting G g gram GA gauge gal gallon GIG gigabyte GND ground H hd head HeCd helium cadmium HeNe helium neon Hg mercury ht height HV high voltage high volume Hz Hertz IA aperture current 2 ABBREVIATIONS i d inside diameter ID identification in inches I O input output IR infrared K K constant thousand kilohm KB kilobytes kg kil
188. ATTENTION is in bold faced text in the printed manual and is red in the online manual Note A Note contains information that is important to remember or helpful in performing a procedure Conventions This manual uses the following conventions to make the material clearer and more concise An example is given for each explanation Italics indicate screen messages such as Waste Bottle Full e Bold indicates a menu item such as Acquisition Courier font indicates text that you must type such as INSTALL The software path to a specific function or screen appears with the double solid right triangle gt gt symbol between succeeding screen options such as Acquisition Parameter e Whenever keys are to be pressed in succession the keys are printed with a space but no punctuation between them such as F9 J Enter e Whenever keys are to be pressed simultaneously the keys are printed with a plus sign between them such as ctrl ait Delete Links to additional information are in blue and are underlined in the online manual To access the linked information select the blue underlined text such as Conventions 1 INTRODUCTION MANUAL DESCRIPTION 1 1 4 PN 4237003 INTRODUCTION SAFETY PRECAUTIONS 1 2 SAFETY PRECAUTIONS This section covers safety precautions that you must take whenever you work on the Elite Additionally when performing a procedure always follow any safety precautions included in th
189. Across Forward Scatter Detector 4 12 7 4 12 13Flow Cell Z Axis Knob Location 4 12 8 4 12 14 PMT2 High Voltage Setting 4 12 9 4 12 15 PMT4 High Voltage Setting 4 12 10 4 12 16 Fluorescence Pickup Lens 4 12 10 4 12 17 Focus Knob Location 4 12 11 4 12 18PMT5 High Voltage Setting 4 12 12 4 12 19 Focus Knob Location 4 12 12 4 13 1 CPU Card Jumper Locations 4 13 1 7 2 1 Acquisition Troubleshooting Flowchart 7 2 2 7 2 2 Filter Configuration 7 2 3 7 3 1 Fluidics Troubleshooting Information 7 3 3 7 4 1 Troubleshooting Fluidics for No Sheath Flow 7 4 1 7 5 1 Troubleshooting Sort Problems 7 5 1 7 5 2 Satellite Droplets 7 5 8 8 2 1 Elite Cytometer Right Panel Removed See Table 8 2 1 8 2 1 8 2 2 Workstation See Table 8 2 2 8 2 2 8 2 3 Sheath and Waste Compartment See Table 8 2 3 8 2 3 8 2 4 Pneumatics Cabinet See Table 8 2 4 8 2 4 8 2 5 Pneumatics Drawer See Table 8 2 5 8 2 5 8 2 6 Pneumatics Panel Rear View See Table 8 2 6 8 2 6 8 2 7 Dual CRT and Connector Panel See Table 8 2 7 8 2 7 8 2 8 Card Configuration Elite See Table 8 2 8 8 2 8 8 2 9 Card Configuration Elite ESP See Table 8 2 9 8 2 9 8 2 10 Cable Interconnections Elite and Elite Analyzer See Table 8 2 10 8 2 10 8 2 11 Multibus Card Cage All Configurations See Table 8 2 11 8 2 11 8 2 12 Data Acquisition Card Cage Elite ESP See Table 8 2 12 8 2 12 8 2 13 Cytometer Inside Rear View See Table 8 2 13 8 2 13 8 2 14 Cytometer R
190. Amp SW R card 18 6705694 Sort Oscillator R2 card 4 6705246 3 PMT Sub Amp SW R2 card 19 6706034 Sort Delay R3 card 5 6705246 3 PMT Sub Amp SW R1 card 20 6704147 Interface and Scaler card 6 6704008 PMT Gated Amp card 21 6704186 Bitmap and Sort Decision card 6706232 Bitmap and Sort Decision R2 card 7 6704904 Quad 20 40 60 Microsecond Delay 2 22 6704185 Prism and Sort Window Test card card 8 6704241 Quad 7 Microsecond Delay 2 card 23 6704142 Data Lister Out card 9 6704241 Quad 7 Microsecond Delay 1 card 24 6705400 Peak ADC and PSH Control card 10 16704904 Quad 20 40 60 Microsecond Delay 1 25 6704169 Quad PSH 2 card card 11 6704009 Scat Aux Gated Amp card 26 6704169 Quad PSH 1 card 12 6705246 3 PMT Sub Amp SW R 2 card 27 6704164 Mux and Scope Interface card 13 6705369 Peak Scatter Mux SW R card 28 6705954 Pulse Pileup Det TOF card 14 16705252 Scat CV SW R1 card 29 16784913 Gated Amp card data acquisition 15 16704913 Gated Amp Extender card data 30 16704907 Pulse Generator and Clock card acquisition 8 2 8 PN 4237003C PARTS LISTS ILLUSTRATED PARTS Figure 8 2 9 Card Configuration Elite ESP See Table 8 2 9 38 37 36 35 34 33 32 77 9 10 11 12 13 14 15 16 17 18 19 20 21 22 7003156 Table 8 2 9 Elite ESP Card Configuration Detail of Parts See Figure 8 2 9 Part Number Descripti
191. Argon laser 90 degree dichroic adjustments to align for maximum co linear signals where both Argon and HeNe signals appear on top of each other on both traces of the Digiscope Close the Argon laser output shutter Open the UV laser output shutter ATTENTION Do not change any flow cell or beam shaper alignment otherwise you will undo the alignment that was just completed in step 13 18 19 20 Align for maximum co linear signals on both traces of the Digiscope using the 90 degree mirror only Open the Argon laser output shutter Block each laser off one at a time with a target or opaque object Observe the change in displayed signals on the Digiscope Note Now that all three lasers are active be sure all three resultant signals are occurring in the same time frame This means the signals should be co linear where all three appear to be on top of each other Time Space Separated Laser Alignment Procedure 1 2 10 Close the Argon laser output shutter Set the UV laser power up or down to create a distinguishable difference in amplitude between the HeNe and UV Laser resultant signals Install the adjustable beam translator between the HeNe laser and its adjustable dichroic filter Note Allow enough space between the translator and the optical rail to mount a single laser target Verify that the a Translator knobs face away from laser b Translator window is perpendicular to the optical bench Adj
192. Be careful not to touch the lens or quartz portion of the flow cell IMPORTANT Risk of inconsistent sheath flow and inconsistent stability If the tubing coming out of the flow cell body is bound or kinked sheath flow and stability inconsistencies may occur Ensure the tubing is not bound or kinked 8 10 11 12 13 14 15 Check the sheath and flush tubing on the two ports on the flow cell body and verify the tubing a Does not bind or kink b Isfree and clear of the cables and the bezel Remove the beam shaper and align the flow cell so the reflection back to the laser interlock is just slightly off to one side Replace the beam shaper Run fluorospheres and align the system Use the flow cell and beam shaper adjustment knobs to ensure that the new flow cell is optimally aligned with the maximum fluorescence and forward scatter intensity possible and that half peak CVs are as low as possible at least under 2 096 After acceptable alignment press SHEATH to stop the beads from running out Enter standard values for the sort set up functions a Set Drive Frequency to 28 32 kHz b Set Crystal Drive to 65 75 c Set Deflection amplitude to 75 8596 Place the ground plate between the bottom of the beam shaper lens and the bottom of the beam shaper itself Note You can vary this a bit as long as it is not too high to cut off the laser signal into the flow cell Set up the camera to view the droplets
193. CH VALVE MANIFOLD MOUNTING ARRANGEMENT OF THE VALVES ON THE MANIFOLD 7003047 4237003C 2 9 3 INSTRUMENT DESCRIPTION PNEUMATICS SUBSYSTEM 2 9 4 Circuit Cards Pneumatic Interface Card There are two communication pathways to the Pneumatic Interface card from the Multibus CPU card The first pathway is a direct cable connection between the Multibus CPU card and the Pneumatic Interface card The Pneumatic Interface card buffers these signals into the appropriate 24 V drive levels to operate the solenoids The second pathway goes from the Acquisition card cage through the Sensor Interface card This pathway allows the CPU to issue requested pressure to the Pneumatic Interface card and to receive information from the pressure sensors and switches The Pneumatic Interface card Controls all pneumatic functions Controls the solenoids The card converts the logic level signals received from the CPU to 24 V levels that drive the fluidic solenoids Note relay activated by the watchdog signals from the Sensor card controls the 24 V Controls the pressure The card receives the requested sheath and sample pressure from the Multibus CPU card via the Sensor card After the values are received the card achieves and maintains the requested pressures Each system uses one of two types of pressure regulators and Pneumatic Interface cards Refer to the chart below Card Used With Reference Pneumatic Interface
194. CK Gain Adjust 10 Sort Output 2 R84 Binary Voltage Step Counter Generation lil Q VW Binary to MN 1 of 10 AA N Decoder ea ANN 7003049A 2 8 10 PN 4237003C INSTRUMENT DESCRIPTION SORT SUBSYSTEM The logic circuits examine the sort event information and the controls settings to either generate e A sort window envelope Figure 2 8 7 of the user preprogrammed length 1 2 or 3 drops No expected sort envelope because an abort condition has occurred abort A sort window envelope longer than preprogrammed sort extension or Asort window envelope shorter than preprogrammed truncation Figure 2 8 7 Sort Envelope TWO DROP SORT ENVELOPE ONE DROP SORT ENVELOPE THREE DROP SORT ENVELOPE 7256011A Function The Sort Oscillator card e Generates various timing signals for sorting A crystal oscillator provides a stable time base A variable digital divider rate multiplier divides the clock to generate the droplet breakoff clock frequency The clock times sorting using circuits on this card and on the sort Delay card The digital clock also creates a synthesized sine wave signal that drives the bimorph This signal goes to the Sort Output card for amplification The strobe drive signal is also derived from this clock second programmed divider generates the FL test frequency e Processes the delayed sort signals received from the Sort Delay card The Sort Osc
195. COULTER EPICS Elite Flow Cytometer am Service Manual RR ee SI 4237003 December 2001 Beckman Coulter Inc Miami Florida 33196 2500 USA LEGAL NOTICES Beckman Coulter Inc makes no representation that upon furnishing this service manual the holder of the manual will have the necessary technical capabilities and know how to properly troubleshoot and repair any of the equipment specified in the manual Beckman Coulter Inc assumes no liability whatsoever including consequential and incidental damages resulting from improper operation of Beckman Coulter instruments after maintenance of Beckman Coulter instruments has been performed by persons not employed by Beckman Coulter Inc Furthermore Beckman Coulter Inc assumes no liability whatsoever for any personal injury or property damage resulting from maintenance and or repair of Beckman Coulter instruments performed by persons not employed by Beckman Coulter Inc READ ALL PRODUCT MANUALS AND CONSULT WITH BECKMAN COULTER TRAINED PERSONNEL BEFORE ATTEMPTING TO OPERATE INSTRUMENT HAZARDS AND OPERATIONAL PRECAUTIONS AND LIMITATIONS WARNINGS CAUTIONS and IMPORTANTS alert you as follows WARNING Can cause injury CAUTION Can cause damage to the instrument IMPORTANT cause misleading results Beckman Coulter Inc urges its customers to comply with all national health and safety standards such as the use of barrier protection This may inclu
196. CPU 1 0 commands Control Card 1 PMT signals 2 The signals EE Ke EE 1 shown are 5 1 M as they would Le PROGAMMABLE BACKPLANE be set for e ra ATTENUATOR wx ditat zs 5 operation These attenuators along with the 10X amps on the Scat Amp Gated Amp Card Ce provide the 1 10 gain function Hie SENSOR A OUT for the AUX channels PTO PMB l ye g MM SENSORSOUT P J11 re PMT4 PMT4 DC RESTORE IN i J7 SENSOR 6 OUT ns DCRESTORE Tum IN 70030042 42370036 2 12 5 INSTRUMENT DESCRIPTION CARD DESCRIPTION GATED AMPLIFIER CONFIGURATION 20 40 60 Microsecond Delay Card The 20 40 60 Microsecond Delay card delays the incoming pulse for a selectable delay of 20 40 or 60 microseconds by selecting one two or three cascaded 20 microsecond delay lines The gain of each 20 microsecond delay section is individually adjustable to ensure unity gain See Figure 2 12 4 Figure 2 12 4 Block Diagram 20 40 60 Microsecond Delay Card DELAY IN INPUT J1 J2 BUFFER 44 25 J2 J7 J8 AMPLIFIER AMPLIFIER AMPLIFIER J10 J11 Note The x second input is used to tee 20 20 20 the signal to
197. E RELAY This cable is part TERMINAL BLOCK of relay panel ON RELAY PANEL This cable is part BLACK STAYCON BROWN SPACE CONNECTORS 24 VOLT GROUND Connected to terminal block 11 TB11 5 24V TB11 11 CND When looking into unit from the back this will be the upper left TB Be sure to match color code J2 J3 CARD EDGE of relay panel assembly EN J1 CARD EDGE Connections to be added 3 7 4 DUAL LASER CONTROL CARD FRONT VIEW 7003004A PN 4237003C 9 INSTALLATION PROCEDURES OPTIONAL LASER INSTALLATION Install the Argon laser as shown in Figure 3 7 3 a Move the Argon laser b Install the rail on the optical table c Connect the new external blower Figure 3 7 3 Argon Laser Installation LASER INTAKE DUCT SCREW X i ARGON CN DUCT PLENUM AIR DUCT NEA A HeNe LASER ARGON e MOUNTING UPRIGHT i e BEAM SPLITTER s 5 HeNe 633 11 8 BEAM SPLITTER gd SPRING BEAM REDUCER EXPANDER 4 PN 4237003C ARGON 438 el L A 10 11 12 13 14 1246 MOUNTING l FEET ep d fl a Ne LASER CABLE WIRE TIE b OPTICAL TABLE VIEWED FROM REAR MOUNTING 7003155A PLATE Target the HeNe laser and air cooled Argon lasers a Ins
198. E 256 SORT UPGRADE 3 22 2 6 10 11 12 13 14 15 16 1T 18 If the Cytometer does not have a printer a At the A prompt B prompt if the system contains two floppy drives type README DOC MORE then press Enter b Review the readme doc file before installing the Elite Software c step 8 If the Cytometer is equipped with a printer a At the A prompt B prompt if the system contains two floppy drives type PRINT README DOC then press Enter b Review the printed readme doc file before installing the Elite Software At the A prompt B prompt if the system contains 2 floppy drives a Type INSTALL then Enter b Follow the instructions on the screen to install the Elite Software When the installation of the Elite Software is complete a From the Main screen select File Open a Select a protocol then click on Ok Press then Enter When the error message UNABLE TO RECEIVE CYTOMETER PROGRAM VERSION PLEASE ACKNOWLEDGE appears press again When the message DO YOU WISH TO TRANSFER THE CONTROL FILE appears press Y then Enter On the Cytometer right monitor the number 1 appears Wait for this number to sequence from 1 to approximately 425 This process will take several minutes If the Elite is equipped with the Autoclone sorting option a When the Control File is transferred the message PLEASE POWER UP THE VALVES AND CALIBRATE THE AUTOCLONE appe
199. E AND REPAIR PROCEDURES PNEUMATICS SYSTEM 4 5 4 Non Low Bleed Regulator Adjustment Procedure Purpose ATTENTION Do this procedure for non low bleed regulator instruments only This procedure applies only to systems with the non low bleed old type regulators This procedure permits minor field alignment to allow for matching the Pneumatic Interface card to the system upon card or sensor replacement This procedure assumes the card has been previously factory adjusted Do this procedure after replacement of the Pneumatic Interface card either regulator or the sheath pressure sensor The following definitions appear in this procedure Set pressure is the bottom pressure displayed on the Control screen which is the pressure entered by the operator in the Service valve screen Read pressure is the upper pressure displayed on the Control screen which is the pressure sensed by the system Gauge pressure is the pressure shown on the external gauge Tools Supplies Needed 0 15 psi gauge Procedure 1 Ifthe card is not known to be system tested factory adjusted make the following adjustments Pot Number Test Point Voltage R31 TP5 7 00 R34 1 3 00 R51 TP2 8 00 R54 TP3 10 00 2 Display the set and read pressures at the Valves screen on the Cytometer and verify that the sheath bottle is filled to the 2200 mL mark 3 Connect the external 15 psi gauge to the sample pressure line with a Y fitting at the sample
200. E AND REPAIR PROCEDURES SORT SENSE FLOW CELL ADJUSTMENT AND VERIFICATION Replacing Metal Fittings with PVC Fittings l Verify that fittings marked 2 and 4 in Figure 4 11 1 are PVC not metal fittings a Ifthe fittings metal replace with PVC fittings b Ifthe fittings are PVC but the flow cell body exhibits a cloudy blackish greenish substance near the point where the insertion rod enter the flow cell body there may be a defective PVC fitting causing a voltage leak during sorting and electrolyzing the insertion rod Go to step 2 c Ifthe fittings are PVC and there is no cloudy blackish greenish substance go to step 3 Figure 4 11 1 Fittings on Valve Assembly Bracket CHECK VALVE e 0 08210 TUBING FF1 SN S 6 FF3 LS UPPER BUBBLE CHECK VALVE H CHAMBER 0 156 1 0 TUBING H i 4 D ri T FITTING Ca gt gt CHECK VALVE 1 0 187 1 0 BRASS z ne vac W 15 gt LOWER BUBBLE CHAMBER SHEATH PRESSURE TRANSDUCER P TRANSDUCER MOUNTING BRACKET 7003188 Regarding Figure 4 11 1 Item 115 a large metal barbed fitting 0 115 i d to 0 180 id Item 2 is a large PVC barbed fitting 0 115 i d to 0 180 i d e Item 3 is a small metal barbed fitting 0 062 i d Item 4 is a small PVC barbed fitting 0 062 i d 4 11 2 PN 4237003C SERVICE AND REPAIR PROCEDURES 4 SORT SENSE FLOW CELL ADJUSTMENT AND VERIFICATION 2 Locate and replace the defective PVC fitting
201. EDURES PREINSTALLATION CHECK The specifications in Table 3 1 2 include the two 7 6 m 25 ft of 16 mm 5 8 in 1 4 hose supplied with each laser However these specifications do not include a 60 p water filter which when new introduces a 5 kPa 0 75 psi pressure delta at 22 7 liters 6 gallons per minute or a 2 kPa 0 30 psi pressure delta at 8 3 liters 2 2 gallons per minute Drainage Drainage must be able to accommodate the water from lasers if any Environment The room where the Elite is to be installed must meet the specifications in Table 3 1 3 Table 3 1 3 Environmental Requirements Description Specification Humidity 0 90 non condensing Heat dissipation into room 2 300 W 7 850 Btu hr with one air cooled Argon laser 2 400 W 8 191 Btu hr with one air cooled Argon laser and one water cooled laser 2 400 W 8 191 Btu hr with one air cooled Argon laser and one Omnichrome Model 56 HeCd laser CAUTION Risk of instrument damage For systems with water cooled lasers the cooling water temperature must not fall below the ambient dew point otherwise condensation forms inside the power supply and may cause catastrophic damage In some situations it may be necessary to provide humidity control for the laser environment Ambient temperature 18 29 64 85 F Stability rate of change Temperature fluctuations within the ambient temperature range can affect sorting performance For optimal sortin
202. ELITE EXE and Cytometer files which the INSTALL program copies over the version 4 0 files Unchanged files are not overwritten Newly manufactured instruments beginning at serial number W48072 are shipped with the software upgrade already installed Tools Supplies Needed PN 42370036 Elite Software Version 4 01 Upgrade PN 6913114 0 US Country use only Elite Software Version 4 01 Upgrade PN 6913060 7 International Use only 3 19 1 INSTALLATION PROCEDURES SOFTWARE VERSION 4 01 INSTALLATION 3 19 2 Installation Procedure ATTENTION Customers can install version 4 01 l Verify that a Elite Software Version 4 0 is already installed on the instrument See Heading 3 18 SOFTWARE VERSION 4 0 INSTALLATION for installation instructions b Watchdog Timing has been completed See Heading 3 17 WATCHDOG TIMING Power up the instrument Press to exit to DOS Insert the 3 5 in disk in the 3 5 in floppy drive Print the README DOC file for additional information SE ee a Verify that the printer is connected and ready to print If your 3 5 in floppy drive is drive A 1 Type A PRINT README DOC P and press Enter 2 Type A INSTALL and press Enter Ifyour 3 5 in floppy drive is drive B 1 Type B PRINT README DOC P and press Enter 2 Type B INSTALL B and press Enter 6 Transfer the control file a When the software screen appears press b Press YJ to transfer the file
203. Education Center Training 3 1 1 Cartons Received 3 1 1 Power Requirements 3 1 1 Water 3 1 2 Drainage 3 1 3 Environment 3 1 3 Space and Accessibility 3 1 3 Optional Printer 3 1 4 Labels 3 1 4 Supplies 3 1 4 INITIAL SETUP 3 2 1 Unloading and Moving the Instrument 3 2 1 Dual CRT Installation 3 2 2 Instrument Installation 3 2 4 CYTOMETER CHECK 3 3 1 CONNECTING ASSEMBLIES 3 4 1 Electronic Cable Connections 3 4 1 Tubing Connections 3 4 3 Options Connections 3 4 4 Laser Option 3 4 4 Optical Drive Option 3 4 4 Printer Option 3 4 4 TESTING ASSEMBLIES 3 5 1 Cytometer Control Computer 3 5 1 Compressor Module 3 5 2 Pneumatics 3 5 2 Camera 3 5 3 3 6 3 7 3 8 3 9 3 10 Lasers 3 5 4 Argon Laser 3 5 4 HeNe Laser 3 5 4 Optional Lasers 3 5 4 Computer 3 5 4 Alignment Check 3 5 4 SYSTEM TESTING 3 6 1 Acquisition Check 3 6 1 Analysis Check 3 6 1 Sort Check 3 6 1 Drive Frequency 3 6 3 Phase Adjustment 3 6 3 Delay Calculation 3 6 5 Monitoring the Sort 3 6 7 PART B OPTIONS AND UPGRADES INSTALLATION OPTIONAL LASER INSTALLATION 3 7 1 Purpose 3 7 1 Tools Supplies Needed 3 7 1 Procedure 3 7 1 HOUSE AIR INSTALLATION 3 8 1 Procedure 3 8 1 GATED AMPLIFIER UPGRADE 3 9 1 Purpose 3 9 1 Tools Supplies Needed 3 9 1 Procedure 3 9 2 FALS Adjustment Assignment and Adjustment Potentiometer Location 3 9 10 PMTI Adjustment 3 9 12 AUX 1 Adjustment
204. Elite water cooled laser options the jet in air flow cell is used for high speed sorting It is available in the following sizes e 51 76u e 1004 e 1504 e 2004 2 3 10 4237003 INSTRUMENT DESCRIPTION 2 OPTICAL COMPONENTS When using the jet in air flow cell an obscuration bar must be placed in front of the fluorescence pickup lens Fast Quartz Flow Cell The 76 high velocity flow cell incorporates the high signal to noise characteristics of sense in quartz design of the sort sense with the sorting speed advantages of the jet in air design The interrogation point is within the 76 p cuvette which minimizes the interference from stream perturbations See Figure 2 3 7 Figure 2 3 7 76 High Velocity Quartz Flow Cell LASER INTERSECTION POINT 7256003 Optical Filters The Elite uses optical filtering to select bandwidths corresponding to specific fluorochromes The optical block allows you to view the filter configuration at a glance and easily change the filter configuration See Figure 2 3 8 PN 4237003 2 3 11 INSTRUMENT DESCRIPTION OPTICAL COMPONENTS 2 3 12 Figure 2 3 8 Optical Filters Location XI XY x x CED 5 c N I Lag veer 5904023A Each filter holder is clearly labeled to identify the filte
205. F OFF Note If paper size is A4 then the statement should appear in front of PAPER LETTER and PAPER ISOA4 should not contain a rem statement 12 Save the changes by pressing E Enter 13 Reboot the computer by pressing ctrl At betete Printer Test l Selecta histogram and press F3 2 Verify that the printout reflects the histogram colors displayed Note The printout may take a while to print depending on the resolution selected 3 20 6 PN 4237003 INSTALLATION PROCEDURES PC MODEL PENTIUM 133 MHz UPGRADE 3 21 PC MODEL PENTIUM 133 MHz UPGRADE Purpose Use this procedure to upgradselecte a workstation computer to 90 MHz Pentium for the following Elite models e 2356 Elite ESP with Gated Amp and or Autoclone Sorting Option and AL 2357 Elite ESP with Gated Amp and or Autoclone Sorting Option 2358 Elite ESP e 2360 Elite with Gated Amp and or Autoclone Sorting Option e 2362 Elite See Table 3 21 1 for the DTX Pentium computers technical specifications Table 3 21 1 Technical Specifications of the DTX Pentium Computer Item Specifications Processor Intel Pentium processor 133 MHz Ports 2 serial ports async 9 pin 160550 compatible 1 parallel EPP port Memory 16 megabyte standard upgradable to 128 megabyte 4 72 pin SIMM sockets 70nS or faster with parity x36 Cache 256k on board second level dire
206. From To Mux and Scope Gated Amp Control R3 slot 1 J25 J2 Mux and Scope Gated Amp Control R3 slot 1 J31 J6 Interconnections from Sensor Area FALS Scat Aux Gated Amp slot 11 J2 PMT1 Scat Aux Gated Amp slot 11 J3 PMT2 PMT Gated Amp slot 6 J4 Gated Amp slot 6 J5 PMT4 PMT Gated Amp slot 6 J6 PMT5 If present PMT Gated Amp slot 6 J7 Bundle these cords and continue 25 Remove the Quad 20 40 60 Microsecond Delay card from slot 10 and put into the extender 26 At the Cytometer a Go to the Scope screen and set TIME DIV to 10 msec div b Go to the Sort screen and set the Frequency to 2 kHz and turn Strobe on c Open the detector compartment and remove the 488 BP filter d Set the Lower Trace to PMT1 INT and the Upper Trace to FALS INT 27 Move the deflection body all the way up and remove the FALS filter 28 Adjust the PMTI gain to 10 and the HV gain for about a three division high PMTI pulse 29 Adjust the FALS INT gain and HV to produce a three division high FALS signal The FALS signal should be clean on the upper trace 30 Close the detector compartment door 3 9 8 PN 4237003C INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE 10sues sseJd eDueu2 0 Aejap Josuas juaJuno s ejdsip yag aures BY je Nd BY
207. GAIN CSC E J6 Filii FL2 LIN OUT J8 LOG AMP EXER He TIE LOG OUT DECODE COMMANDS Deni e FROM BACKPLANE CPU AND CONTROL CARD FUNCTIONS P1 BACKPLANE CONNECTOR 1003024 DUAL FL AMP NON SWITCHAMPS PN 4237003C 2 11 3 INSTRUMENT DESCRIPTION CARD DESCRIPTION NON GATED AMP NON SWITCH CONFIGURATION EXCEPT ANALYZER 2 11 4 PN 4237003C CARD DESCRIPTION GATED AMPLIFIER CONFIGURATION INSTRUMENT DESCRIPTION 2 12 CARD DESCRIPTION GATED AMPLIFIER CONFIGURATION Gated Amp Control Card The Gated Amp Control card provides an interface and cable connection between the DT Interface card and the Gated Amp backplane upper backplane The card recognizes the range of port addresses that correspond to cards in the upper backplane and puts the proper addresses and data words on the backplane See Figure 2 12 1 Figure 2 12 1 Block Diagram Gated Amp Control Card LATCH FOR DISCRIMINATOR VALUE AND D TO A CONVERTER TP3 The trigger source is J4 gated by the comparator J2 PMT TRIGGER a so trigger gt disc level to TO SOURCE TRIGGER LOG be displayed MUX AND GAIN AMP SCOPE J3 e FOR SCAT AUX 4 _ DISPLAY URGE Select scat aux Compare trigger signal
208. H Parallel Port Address 378 H Press to return to the Setup menu Select Write to CMOS and EXIT Press Y Enter PN 4237003C 4 9 PN 42370036 SERVICE AND REPAIR PROCEDURES SOFTWARE INSTALLATION SOFTWARE INSTALLATION Purpose This procedure provides general software installation procedures Always follow the instructions provided with the software package for specific installation instructions This procedure affects the following serial numbers low ORSO9119 high V39054 Tools Supplies Needed Software Kit PN 6912816 Procedure 14 15 16 1T Install the Elite Software diskette 1 in drive B At type B Install Press and follow the instructions provided Select the customer s Printer type when prompted Select Standard and press any key when prompted to do so until install is finished and prompt appears Turn Workstation pc OFF and wait 10 seconds Turn Workstation pc ON and allow system to boot Install PrintQ diskette in drive B If in Elite Software press to exit Type B Install Press Enter Before You Can Use PRINTQ 6 to Make Printing Fly appears on the screen Press Enter Answer the following questions by typing the correct response and pressing To accept the response that appears press Ente Install from Which Drive Type B and press Enter Boots from Which Drive Type C and press Enter Install on Which Drive Ty
209. ION PROCEDURES ENTERPRISE LASER UPGRADE 3 14 4 18 19 20 21 22 23 24 Remove all targets 17 Block all beams except the 488 and align the flow cell and beam shaper using fluorescent particles Adjust the beam focus for narrowest peak fluorescence Block the 488 beam and allow the UV beam to enter the flow cell Note You can monitor the beam s location by holding a business card between the flow cell and scatter sensor Make slight adjustments to the optical rail UV beam mirror until you see a scatter or GFl signal Adjust for optimum amplitude Compare the UV beam and 488 beam peak pulse width Adjust the UV beam width on the beam separator to match the 488 pulse width This disturbs the UV beam alignment compensate as needed Block the 488 and UV beams and adjust the HeNe laser mirror for peak scatter Connect the laser interlock cable between the laser power supply and the heat exchanger interlock cable Note If the system operates with tap water connect the laser interlock cable between the laser power supply and the heat exchanger interlock defeat Verify that the Enterprise laser shuts down when the Laser ON OFF key switch on the Elite system is turned to OFF or when the laser compartment door is opened Replace all covers PN 4237003C INSTALLATION PROCEDURES COLLECTION OPTICS UPGRADE 3 15 COLLECTION OPTICS UPGRADE PN 4237003C Purpose Use this procedure to upgrade flow cell
210. J6 EE NON CEN RR d NORM Re T lt 1 TOF NE gt l BEM WIDTH TME NORMAL 5 DELAY gt IZATION VOLTAGE CONVERSION RAW Ww PULSE WIDTH ADJUSTED PULSE WIDTH 7003060 The output voltage of the card is proportional to the length of time that the pulse exceeds 1496 of its own amplitude The stretched pulse output from PSH8 goes to a resistor voltage divider to develop the 14 reference level that is applied to one input of a voltage comparator The comparator initially sees the reference level on its positive input and outputs a high As the delayed input pulse rises above the reference level the comparator output goes low until the input pulse falls back below the reference level 2 8 16 PN 4237003C INSTRUMENT DESCRIPTION 2 SORT SUBSYSTEM As the particle enters the laser beam the signal pulse rises to its peak value it does not drop until the particle leaves the laser beam Since each particle in the sample flows through the laser at the same speed the card actually measures the diameter of the particle by measuring how long it takes that particle to pass a certain point However since the laser beam has a finite diameter the pulse length at the comparator output actually represents the time it takes the particle to pass through the beam And si
211. J7 SHEATH INLINE J2 939 JTO PRESSURE SENSOR J9 ji SHEATH EMPTY SWITCH WASTE FULL SWITCH PSU RINSE EMPTY SWITCH LEFT RIGHT SYSTEM PRESSURE SWITCH CRT CRT Joo O CHAMBER FILTER CLOGGED SWITCH MEE WASTE FILTER CLOGGED SWITCH J2 d J2 DUAL CRT jg SEN j 90V CONTROL 78 Lou 490 a REM Er _ 36 V REGULATOR DUAL LASER m LASER 90 36 V REGULATOR e CONTROL i CONTROL a CONNECTORS 36V REGULATOR CARD 0 WIDTH POWER FRONT BACK PORCH XSISTORS SWITCHING POWER EN SUPPLY e n n K Jb i IIN 39335858 53859383 58555355 55555995 PULSE QUAD QUAD ADC amp PSH Tus BIWAR INTERFACE SORT SORT SORT m ux GENERATOR PPU SCOPE PSH 1 PSH 2 CONTROL DUT DECISION amp SCALAR DELAY OSCILLATOR OUTPUT 58 AE TEST s T ESP UNITS ONLY SEE SEPARATE SHEET WORKSTATION COMPUTER LOWER BACKPLANE amp T PN 4237003 2 4 3 INSTRUMENT DESCRIPTION 2 4 4 2 4 4 PN 4237003 INSTRUMENT DESCRIPTION CONTROL AND DISPLAY SUBSYSTEMS 2 9 CONTROL AND DISPLAY SUBSYSTEMS Multibus Subsystem The Multibus subsystem is the control computer for the Cytometer The Multibus is an industry standard backplane that provides the necessary Address lines Data lines Timing lines e Control lines These lines allow the various cards attached to the multibus to function as a computer system Although many lines such as a
212. K DEFEATED AVOID EYE OR SKIN EXPOSURE TO DIRECT OR SCATTERED 12 WATTS MAX CW RADIATION ARGON ION LASER 5965003B PRACTICAL LIMITS CLASS IV LASER PRODUCT Figure 1 2 3 Laser Warning Labels Helium Cadmium Laser INVISIBLE LASER RADIATION AVOID DIRECT EXPOSURE TO BEAM LESS THAN 50 0 MW CLASS HeCd LASER PRODUCT DANGER LASER LIGHT WHEN OPEN AVOID DIRECT EXPOSURE TO BEAM INVISIBLE RADIATION IS EMITTED FROM THIS APERTURE AVOID EXPOSURE 59650228 1 2 4 4237003C INTRODUCTION SAFETY PRECAUTIONS Figure 1 2 4 Laser Warning Labels Omnichrome Helium Cadmium 74 Laser INVISIBLE LASER RADIATION AVOID DIRECT EXPOSURE TO BEAM LESS THAN 50 0 MW CLASS HeCd LASER PRODUCT INVISIBLE RADIATION IS EMITTED FROM DANGER THIS APERTURE LASER LIGHT WHEN OPEN AVOID DIRECT EXPOSURE TO BEAM AVOID EXPOSURE ETT Figure 1 2 5 Laser Warning Labels Coherent Innova Enterprise Laser DANGEROUS VOLTAGE UNDER THIS COVER VISIBLE AND INVISIBLE LASER RADIATION IS EMITTED FROM THIS APERTURE AVOID EXPOSURE VISIBLE AND INVISIBLE LASER RADIATION AVOID EYE OR SKIN EXPOSURE TO DIRECT OR SCATTERED RADIATION ARGON ION LASER 2 WATTS MAN CW PRACTICAL LIMIT CLASS IV LASER PRODUCT 7003159A 1 2 5 1 INTRODUCTION SAFETY PRECAUTIONS 1 2 6 Figure 1 2 6 Laser Warning Labels Coherent Innova 305 Laser VISIBLE AND INVISIBLE LASER R
213. LASER INSTALLATION Purpose Perform this procedure to install an optional third laser which can be e An Innova 305 laser e Omnichrome 356 laser An Innova 90 laser with remote box Currently Elite systems are shipped already equipped with the appropriate optical components interlock components and covers As such installation is limited to unpacking and mounting the optional laser system However older Elite systems with optional lasers require installation of the cover kit optics kit and appropriate laser mounting kit Table 3 7 1 lists possible laser configurations Table 3 7 1 Possible Laser Configurations Standard Plus Standard Plus Standard Plus One Water Cooled U V 1305 One Water Cooled U V 90 5 One HeCd U V Series 74 Standard Model Model 1 air cooled Cyonics 1 air cooled Cyonics 488 laser 1 air cooled Cyonics 488 laser 1 air cooled Cyonics 488 laser 488 laser 1 HeNe 633 laser 1 HeNe 633 laser 1 water cooled Innova 305 351 1 Coherent laser 1 HeNe 633 laser 1 water cooled 90 5 351 1 Coherent laser 1 HeNe 633 laser 1 HeCd U V Multimode 325 Omnichrome laser PN 4237003C Tools Supplies Needed Kit for laser to be installed Procedure WARNING Risk of personal injury Laser manufacturers of certain accessory lasers may require factory certification of the installer Do not install any laser unless you have the required training and or certification WARNING Risk of persona
214. LUSTRATED PARTS Item Part Number Description 1 N A Vertical rotation knob 2 N A Horizontal rotation knob 3 3814223 Adapter 45D mirror 4 N A Height setscrew 5 6857634 Beam splitter stage 6 N A Rail setscrew 7 N A Fine adjustment setscrew 8 N A Fine adjustment knob PN 4237003C 8 2 29 8 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 30 Beam Shaper See Table 8 2 30 5 7003130A Table 8 2 30 Beam Shaper Detail of Parts See Figure 8 2 30 Item Part Number Description 1 N A Vertical translation knob 2 N A Horizontal rotation knob 3 N A Vertical rotation knob 4 N A Height setscrew 5 N A Rail setscrew 6 N A Fine adjustment setscrew 7 N A Fine adjustment knob 8 N A Horizontal translation knob 9 N A Focus ring 10 N A Lateral translation knob 8 2 30 42370036 PARTS LISTS 8 ILLUSTRATED PARTS Figure 8 2 31 Flow Cell Stage Elite ESP See Table 8 2 31 7003088A Table 8 2 31 Elite ESP Flow Cell Stage Detail of Parts See Figure 8 2 31 Item Part Number Description 1 3814271 Ultra compact positioner 2 1010932 Upper nut insertion rod gland nut 3 6856762 Flow cell body high optical 4 6232208 Hose barb fitting elbow 0 093 i d nylon white 5 6859397 Sort sense 140 u 6859300 Flow cell tip 100 u 3X 6859313 Flow cell tip 76 u 6 6858368 Flow cell bracket PN 4237003C 8 2 31 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 32 Flow Cell Sta
215. Lister Out card Data Taker Interface See Data Taker Interface card Digiscope See Digiscope card digital values reading 2 13 1 DT Interface See Data Taker Interface card Dual CRT Control See Dual CRT Control card Dual FL Amp SW R See Dual FL Amp SW R card Dual FL Amp See Dual FL Amp card Dual FL SW R See Dual FL SW R card Dual Laser Control See Dual Laser Control card Gated Amp configurations used in 2 12 1 Gated Amp Control R3 See Gated Amp Control R3 card Gated Amp Control See Gated Amp Control card HV DAC Control See HV DAC Control card HV DAC See HV DAC card in Cytometer 2 5 6 in Cytometer list of 2 5 6 Interface and Scaler R See Interface and Scaler R card Interface and Scaler See Interface and Scaler card KIR3 See KIR3 card Lister A B See Lister A B card Lister AT See Lister AT card Lister Out See Lister Out card Motor Control See Motor Control card Multibus CPU See Multibus CPU card Mux and Scope Interface See Mux and Scope Interface card Mux and Scope See Mux and Scope card noise isolating input from 2 13 4 non Gated Amp configurations used in 2 11 1 not to be removed 3 3 1 Peak ADC PSH Control card See Peak ADC PSH Control card PN 4237003C Peak See Peak ADC card Peak Scatter Sensor See Peak Scatter Sensor card Peak Scatter Mux SW R See Peak Scatter Mux SW R card PMT Gated Amp See PMT Gated Amp card Pneumatic Interface See Pneumatic Interfac
216. MT2 from PMT3 to 25 b Acquire and observe that PMT3 mean is reduced by 25 512 0 25 512 384 Note Mean for PMT3 should be 384 15 channels c Set PMT3 PMT2 subtract PMT2 from PMT3 to 5096 d Acquire and observe that PMT3 mean is reduced by 50 Note Mean for PMT3 should drop to 256 15 channels Log Amplifiers If there is no log amp output or if the output is incorrect 1 At the amplifier cards either on the backplane or at the card input measure the voltage to ensure that the 15 V power supplies yield 15 00 V a Ifnot adjust the power supply b If adjusting the power supply is not effective replace the power supply Disconnect the amplifier input Locate the correct output connection Note Refer to Chapter Heading SCHEMATICS for schematics and interconnect diagrams or read the card edge to locate the correct output connection Measure the amplifier output it should be negative at least 100 mVdc Note If the card has 0 output or positive output replace the card PN 4237003C TROUBLESHOOTING TROUBLESHOOTING FLUIDIC PROBLEMS 7 7 3 TROUBLESHOOTING FLUIDIC PROBLEMS Purpose Do this procedure to detect and repair fluidic problems Generally fluidic problems are categorized into three groups e Leaks where fluid air or liquid escapes from the system e Blockages where fluid air or liquid flow is restricted Improper functions where fluidic control components such as valves
217. MT2 gain and HV gain for a three division high pulse b Adjust the PMTI gain and HV gain for a three division high pulse At the Gated Amp Control Screen Figure 3 9 5 select FALS LWBM and PMT1 UPPBM Ensure that the delay is set to 7 microseconds at the Gated Amp Control Screen Adjust R107 on the Quad 7 Microsecond Delay card slot 9 for the same amplitude as in step 5 At the Gated Amp Control Screen change the delay to 20 microseconds ATTENTION The following adjustment must be made on the component side of the Quad 20 40 60 Microsecond Delay card slot 10 PN 4237003C 10 11 12 13 14 15 16 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE Adjust R96 on the Quad 20 40 06 Microsecond Delay card slot 10 as above Change the delay to 40 microseconds and repeat as above adjusting R100 Change to 60 microseconds and adjust R104 as above Change the delay to 7 microseconds Set to LWBW Press SHEATH at the Cytometer membrane panel At the Cytometer go to the Options screen and set the Align LED to Pulsed 100 and 2 kHz Remove the 488 blocker the 525 bandpass the 575 bandpass and the 675 bandpass filters ATTENTION Some instruments may not have a 625 DL but a 640 DL which produces a ragged pulse on PMT 5 The 600 DL may be moved out under PMT4 to obtain signals for PMT 4 and PMT 5 17 Verify that the 488 DL is under PMTI 550 DL is under PMT2 600 DL is under PMT3 PN 4237003C and 625 DL is un
218. Main Set the AUX2 peak gain to 10 Adjust the 5 high voltage for 5 V pulse height on trace 2 top trace See Figure 4 12 18 4237003 4 12 11 OPTICAL ALIGNMENT PROCEDURE SERVICE AND REPAIR PROCEDURES Figure 4 12 18 PMT5 High Voltage Setting A 7003192A 4 Obtain maximum pulse heights a b c d Adjust the flow cell horizontal knob as needed Adjust the flow cell vertical knob as needed Adjust the laser beam vertical knob as needed Adjust the fluorescence pickup lens for simultaneous maximum pulse heights Repeat step 4 at least once Obtain minimum pulse widths by adjusting the focus knob See Figure 4 12 19 for the knob location 7 Collect new histograms and recheck the statistics Figure 4 12 19 Focus Knob Location 4 12 12 FOCUS ADJUSTMENT KNOB 7003205A PN 42370036 SERVICE AND REPAIR PROCEDURES EXTERNAL MEMORY CARD REPLACEMENT 4 13 EXTERNAL MEMORY CARD REPLACEMENT Purpose Use this procedure to either replace the 256K Memory card with the External Memory card or to replace a defective External Memory card The circuitry on this External Memory card works effectively with the Cytometer CPU circuitry which minimizes the potential for a CPU lockup to occur Tools Supplies Needed External Memory card PN 7000681 Jumper 2 position discrete wire PN 2121023 Procedure 1 Remove the left panel to allow access to the Multibus card cage in the elect
219. Mounting Hardware Detail of Parts See Figure 8 2 33 8 2 33 Argon Laser Mounting Hardware Detail of Parts See Figure 8 2 34 8 2 34 HeNe Laser Mounting Hardware Detail of Parts See Figure 8 2 35 8 2 35 Laser Placement Top View Detail of Parts See Figure 8 2 36 8 2 36 Cytometer Inside Rear View Detail of Parts See Figure 8 2 37 8 2 37 Omnichrome HeCd Laser See Figure 8 2 38 8 2 38 Pneumatic Interface Card Adjustments A 1 2 Multibus Card Cage Jumpers and Switch Settings A 2 2 Non Gated Amp Card Cage Jumpers 2 2 Gated Amp Card Cage Jumpers A 2 3 Data Acquisition Card Cage Jumpers A 2 3 Line Voltage Jumpers A 2 4 Power Supply Adjustments A 3 2 XXV xxvi PN 4237003 CONTENTS WI INTRODUCTION 1 1 1 1 1 MANUAL DESCRIPTION 1 1 1 Scope 1 1 1 Intended Audience 1 1 1 Organization 1 1 2 Format 1 1 2 Numbering Format 1 1 2 Special Headings 1 1 3 Warning 1 1 3 Caution 1 1 3 Important 1 1 3 Attention 1 1 3 Note 1 1 3 Conventions 1 1 3 12 SAFETY PRECAUTIONS 1 2 1 Electronic 1 2 1 Biological 1 2 1 Laser 1 2 2 Laser Beam Hazards 1 2 2 Radiation Hazards 1 2 2 Notify the Customer About Barrier Protection 1 2 9 ILLUSTRATIONS 1 2 1 Laser Warning Labels Coherent Innova 300 Water Cooled Argon Laser 1 2 3 1 2 2 Laser Warning Labels Innova 90 Water Cooled Argon Laser 1 2 4 1 2 3 Laser Warning Labels Helium Cadmium Laser 1 2 4 1 2 4 Laser Warning Labels Omni
220. Ms Computer IBC 86C 8 2 11 2 6804637 Bertan Supply No 5 card PMT HV Bertan 7000681 External Memory 8 2 9 4 8 2 11 5 PN 4237003C MASTER PARTS LIST IN NUMERICAL ORDER CATEGORY Table 8 1 5 Covers Doors PARTS LISTS Part Number Description Figure Item 1017262 Cover plate bottom HeCd laser 8 2 33 5 1018721 Cover tray Autoclone Sorting Option 8 2 23 11 1018859 Cover pinch valve 8 2 17 4 6856509 Door bottle tray 8 2 3 1 6856889 Door pneumatic side 8 2 4 5 6858097 Door analyzer sample stage Table 8 1 6 Cytometer Part Number Description Figure Item 1010932 Nut upper insertion rod gland 8 2 31 2 8 2 32 2 1018859 Cover pinch valve sample drawer Analyzer 8 2 17 4 1018860 Template FALS detector sheet 1019167 Switch overlay and membrane 8 2 1 16 1019224 Probe pick up 8 2 17 5 1019224 Tube sample sample drawer 8 2 17 5 1019958 Bar obscuration jet in air 8 2 16 3 1020069 Tube holders 12 x 75 8 2 1 22 1020377 Gusset LH rear 8 2 37 1 1020909 Switch membrane Elite Analyzer 1020913 Block sample station sample drawer 8 2 17 12 1022054 Shield splash sample drawer 8 2 17 1 2105057 Power connector 8 2 14 5 2305005 Crystal bimorph 2306042 Clock crystal oscillator 2306046 Transducer sheath pressure 8 2 6 8 2510014 Mount shock 8 2 14 3 2603012 Fan blower rear door fans 79 CFM 115 Vac 8 2 4 4 2603065 Fan blower 112CFM 115Vac 8 2 14 2 8 2 15 1 2802006 Screw obscuration bar 8 2 16 1
221. NSTALLATION 3 16 1 Purpose 3 16 1 Tools Supplies Needed 3 16 1 Installation 3 16 1 Initial Flow Cell Alignment 3 16 2 3 17 WATCHDOG TIMING 3 17 1 Purpose 3 17 1 Tools Supplies Needed 3 17 1 Procedure 3 17 1 3 18 SOFTWARE VERSION 4 0 INSTALLATION 3 18 1 Purpose 3 18 1 Tools Supplies Needed 3 18 1 Procedure 3 18 2 3 19 SOFTWARE VERSION 4 01 INSTALLATION 3 19 1 Purpose 3 19 1 Tools Supplies Needed 3 19 1 Installation Procedure 3 19 2 Waste Bottle Modification for Software Version 4 01 3 19 2 PN 4237003 3 iii 3 20 3 21 COLOR PRINTER UPGRADE 3 20 1 Purpose 3 20 1 Tools Supplies Needed 3 20 1 Printer Setup 3 20 1 Software Installation 3 20 1 Editing CGI CFG File DOS 6 0 and Higher 3 20 2 Editing CGI CFG File Lower Than DOS 6 0 3 20 4 Printer Test 3 20 6 PC MODEL PENTIUM 133 MHz UPGRADE 3 21 1 Purpose 3 21 1 Tools Supplies Needed 3 21 2 Procedure 3 21 2 ELITE 256 SORT UPGRADE 3 22 1 Purpose 3 22 1 Tools Supplies Needed 3 22 1 Procedure 3 22 1 Install the New Bitmap and Sort Decision Card 3 22 1 Install The Elite Software 3 22 1 ILLUSTRATIONS 3 1 1 3 2 1 3 2 2 3 2 3 3 2 4 3 2 5 3 2 6 3 2 7 3 3 1 3 4 1 3 4 2 3 4 3 3 5 1 3 5 2 3 5 3 3 6 1 3 6 2 3 7 1 3 7 2 3 7 3 3 7 4 3 7 5 3 7 6 3 7 7 3 7 8 3 7 9 3 7 10 Warning Labels on Instrument 3 1 5 Unloading the Cytometer 3 2 1 Dual CRT Installation 3 2 3 Rinse Bottl
222. OCEDURES ELECTRONIC ADJUSTMENTS CABLE AND JUMPER CONFIGURATIONS Table 4 10 2 Multibus Card Cage Cards Jumper and Switch Settings Continued Card Jumper and or Switch Settings 256K Memory Note If the instrument was manufactured after 10 30 96 or if the instrument has the new computer card PN 6706571 see the jumper settings for the External Memory card 5 1 SW2 SW3 SW4 SW5 SW6 SW7 SW9 x O O X gt X Closed Open x O 5 O Oo X x O O ze O ze Cc O O O XxX O O O X E19 E20 open E25 E26 jumpered External Memory X1 jumpered X6 a 3 pin jumper jumpered in the 12 V position Camera Interface E19 E20 E21 E22 E37 E39 E38 E42 E41 E45 E44 E46 Dual CRT Control E11 E12 E13 E14 Digiscope E69 70 E31 39 E25 17 1 ALL ON Dual Laser Control E4 E7 E25 E26 E28 E29 E31 E32 E34 E35 E37 E38 E40 E41 Non Gated Amp Card Ca ge Jumper the cards in the Non Gated Amp card cage as shown in Table 4 10 3 Table 4 10 3 Non Gated Amp Card Cage Jumper Setting Slot Card Jumpers 1 left Gated Amp Control R2 E4 E5 E7 E8 7 3 PMT Sub SW R E9 E11 8 Dual FL SW R E8 E9 E11 E12 9 Scat CV SW R E9 E11 E5 E7 10 Peak Scatter Mux SW R E1 E2 E3 E4 E5 E7 16 Sensor Interface E2 E3 E5 E6 E8 E9 E11 E12 E13 E14 E16 E17 E19 E20 E22 E23 Gated Amp Card Cage Jumpe
223. OCUS RING HORIZONTAL TRANSLATION HORIZONTAL ROTATION VERTICAL KNOR FINE me ADJUSTMENT HEIGHT SETSCREW MICRO OPTICAL FINE ADJUSTMENT RAIL SETSCREW RAIL SETSCREW 59950138 The laser beam must pass through the optical center of the beam expander reducer to prevent optical irregularities The beam expander reducer should be installed directly in front of the laser to allow the beam translator and steering optics to be adjusted without having to realign and focus the beam expander reducer each time Side Scatter and Fluorescence Collection Optics Sort Sense Flow Cell Tips Sort sense flow cell is designed for Sense in Quartz sorting using an air cooled laser It is the principal flow cell for the Elite ESP The sort sense contains 250 square flow channel and a 100 um diameter jetting orifice There are two designs the original sort sense Figure 2 3 5 and the new 3x sort sense Figure 2 3 6 PN 4237003C 2 3 9 2 INSTRUMENT DESCRIPTION OPTICAL COMPONENTS Figure 2 3 5 Sort Sense Flow Cell Original X XN e SN FLOW FLOW Que WE 0 CELL MIRROR LENS 7256002A Figure 2 3 6 100 Sort Sense Flow Cell Cut away View SAMPLE INTRODUCTION ROD 2 4 FLOW CHAMBER HOUSING MAGNIFYING _ LENS JETTING n ii ORIFICE 7256010 Jet in Air Flow Cell Tips Available with
224. OLOANNOD 331 Xv09 Wad vLiNd Y MS 14 1vna Ld MS 81 LIN d E e 9015106 NOWO Suv er cbf ONY LEP 1NI 106 ILON AYJd 193 n vir 34098 UMS A9 1V9S YALLWOIS AYJd 5 3311595 Wad WAISAS LINd S e 18je3g xyeag sduy ejqeuaiMs LL 9 Z aunbl4 PN 4237003C 2 5 16 INSTRUMENT DESCRIPTION 2 ACQUISITION SUBSYSTEM Gated Amp Figure 2 6 12 is a block diagram that shows the circuit card signal interconnections for the Gated Amp system The functions that each card performs appear in the individual card descriptions including an explanation of how signals are selected and processed within the cards The illustration traces the FALS and PMT signals from where they enter on the left to where they pass into the Mux and Scope card on the right PN 4237003C 2 6 17 INSTRUMENT DESCRIPTION ACQUISITION SUBSYSTEM 2 6 18 PN 4237003C HAS 1 SCAT AUX TRIG SOURCE IN 52 EACH SYSTEM TRIG TRIG SCAT TRIGGER SRC WINDOW b 8027078 ll 5 S x 200000000 SRC OUT lt 2 USE BNC BULLET COAX 8028121 TRIG SCR IN 0 18 18 Figure 2 6 12 Block Diagram Gated Amplifier SLOT 11 SCAT AUX GATED FLAS OUT Lee E2 E3 OUT FL2 OUT AUX1 OUT FL3 INT AUX20UT FL3 LOG PMT1 OUT FL3 PK TRIG SRC OUT AUX IN AUX2 IN FROM FAST TO FROM SLO
225. ORTRAIT rem PAPER ISOA4 PAPER LETTER PLISTSIZE 4096 rem FF OFF Note If paper size is A4 then the rem statement should appear in front of PAPER LETTER and PAPER ISOA4 should not contain a rem statement PN 42370036 3 20 3 INSTALLATION PROCEDURES COLOR PRINTER UPGRADE 3 20 4 10 11 Save the changes by pressing At E S Exit by pressing At amp E X Reboot the computer by pressing Alt Delete Do Printer Test Editing CGI CFG File Lower Than DOS 6 0 1 2 3 EDLIN and press Enter Type L and press Enter Fight lines followed by a series of printer configurations appears Locate the printer driver without a REM statement before it This should match the printer currently in use if any Deactivate the active printer driver a Type the number of the line that contains the active printer configuration and press The same statement displays above the current cursor line b Press Insert c Type REM and press d Press Enter e If the active printer driver is for a Laserjet a standard dot matrix or an Epson printer driver 1 the number of the Resolution line and press Ente 2 Press Insert 3 Type REM and press 4 Enter f If the active printer driver is for an Epson printer 1 Typethe number of the Printer Epsonx sys line and press Enter 2 Press 3 Type REM and press 4 Press
226. PARTMENT FLOOR 7003124A PN 4237003 3 2 7 INSTALLATION PROCEDURES INITIAL SETUP 3 2 8 PN 4237003C INSTALLATION PROCEDURES 3 CYTOMETER CHECK 3 3 CYTOMETER CHECK ATTENTION Do do remove any of the circuit cards unless instructed to do so Unbolt and remove the two front panels from the Cytometer Verify that all circuit cards are properly seated in the rack Verify that all ribbons and coaxial cables are secure Ne Open the flow cell compartment door and tilt the cover up by removing the two retaining screws at the rear Gi If installing the Elite Analyzer proceed to step 7 6 Remove the pinch valve deactivator clip from the sample line pinch valve See Figure 3 3 1 Figure 3 3 1 Pinch Valve Deactivator Clip One pinch valve deactivator clip located here un 7003002A 7 Verify that the scatter sensor mask is correctly in place PN 4237003C 3 3 1 INSTALLATION PROCEDURES CYTOMETER CHECK 3 3 2 PN 4237003C INSTALLATION PROCEDURES 3 CONNECTING ASSEMBLIES 3 4 CONNECTING ASSEMBLIES Electronic Cable Connections l Verify that the two power switches on the Cytometer are in the OFF position 2 Verify that any optional accessories printers optical drives or lasers are turned OFF and disconnected from any power source 3 Inspect all cables to ensure that the pins are straight and not pushed in Insert the yellow plugs
227. PN 6704552 Norgren pressure regulators Old Type Pneumatic Interface PN 6705504 Low bleed regulators New Type See Heading 4 5 PNEUMATICS SYSTEM for additional information Sample Pressure Systems There are two types of sample pressure systems old type and new type 014 Type With the old type of sample pressure system Figure 2 9 2 after you select the sample pressure the CPU writes the information to the Pneumatic Interface card The card retains this value and applies it to the digital to analog converter DAC You can read the DAC output with a DVM at test point 8 TP8 To confirm operation of all the circuitry to TP8 you can monitor the test point while entering different samples pressure values through the Service Valves screen If you cannot confirm operation of the circuitry there is no need to replace any of the components beyond this point including regulators The exact voltage of TP8 is a function of the DAC reference voltage which you can measure at TP1 and adjust via R34 Therefore if TPI read 3 0 V which is a typical value and the requested sample pressure is 15 psi then the PN 4237003C INSTRUMENT DESCRIPTION 2 PNEUMATICS SUBSYSTEM DAC output is 3 0 V You can proportionally calculate the TP8 voltage for a pressure less than 15 psi 10 psi yields 2 0 V 5 psi yields 1 0 V and so on The closed loop feedback system consists of the following Sample regulator Sample pressure sensor Opamp networ
228. PPLY 15 V 3 4 A SUPPLY 7003068A REAR VIEW 2 8 20 42370036 INSTRUMENT DESCRIPTION PNEUMATICS SUBSYSTEM 2 9 PNEUMATICS SUBSYSTEM General The pneumatics system operates as follows Note For more specific information see the Pneumatic Hydraulic Layout and Pneumatic Hydraulic Layout Analyzer engineering schematics For the part numbers and location of these schematic files see Chapter 6 The Compressor module provides pressure and vacuum to the system The compressor module has one electrical and three pneumatic connections One connection is the air input for the compressor The third connection is a vacuum return The compressor output is connected to the cooling coil located under the tabletop The cooling coil allows the heat that has built up in the compressed air to dissipate and also allows moisture in the compressed air to condense The cooled air next passes through the air water separator where the condensed water is trapped and the dry air passes through Valve 15 water drain periodically removes the water trapped in the air water separator The system pressure switch monitors the air pressure after the air water separator This switch is adjustable and is nominally set to close when the pressure reaches 22 psi it reopens when pressure drops to about 18 psi The air goes to the manifold assembly where it supplies pressure to all the electrically controlled solenoid valves For more information on valves refe
229. PPY DRIVE SEEK AT BOOT FLOPPY DRIVE SWAPPING SYSTEM BOOT UP SEQUENCE SYSTEM BOOT UP SPEED BASE MEMORY SIZE EXTERNAL CACHE MEMORY NTERNAL CACH EMORY FAST POWER ON SELF TEST POST PASSWORD CHECKING VIDEO SHADOW C000 32K SHADOW C800 16K SHADOW CC00 16K SHADOW D000 16K SHADOW D400 16K SHADOW D800 16K SHADOW 16K F000 CACHABL IRQ3 DI I P P I D D 44 PC MODEL PENTIUM 133 MHz UPGRADE PC MODEL PENTIUM 133 MHz UPGRADE Connector on rear of Lister A B card connector located on card MB 3 5 NOT TYPE NOT INSTALLED 30 INSTALLED 1 PRESENT VGA EGA DISA BLED N N N o w WW wW A 3 A N UJ ON ENAB DISA A HIGH 640K WR BACK WR BACK ENAB ED SETUP CACHE SA SA SA SA SA ve te e DJs CJ w w w w Ed SA ENAB INFORMATIVE ONLY NO SELECTION 3 21 3 3 PC MODEL PENTIUM 133 MHz UPGR
230. PROCEDURE PN 42370036 Purpose Do this procedure to install PMT 5 on instruments that have switchable amplifiers Tools Supplies Needed LI Mod Kit 5 Switch Amps 6913056 9 Elite minimum software version 4 01 Analyzer minimum software version 4 01 Bertan Supply No 5 card PN 6804637 8 Analyzer HV DAC card PN 6703941 6 HV coax cable PN 6027024 4 PMT PN 6856833 1 6 32 x 1 5 in thumbscrew PN 2815003 2 PMT 4 cable PN 6028205 6 PMT signal coax cable PN 6028205 6 PMT Sub Amp Amp SW R2 card PN 6705246 3 BNC T connectors 2 PN 2121537 6 BNC bullet coax cables short 4 PN 6027482 7 27 in 69 cm bullet bullet coax cable 3 PN 6027803 2 Hardware Configuration and Installation 1 2 power Remove the top cover rear panel to expose the switcher and front right panel to expose the data acquisition and gated amp card cages Open the rear left door to expose the Bertan card cages Remove the lower Bertan card cage card retainer Install new Bertan Supply No 5 card in fourth card slot from the left including the slot for the HV DAC card in the lower Bertan card cage Configure new Analyzer HV DAC card as HV DAC 2 by jumpering e E5 E9 e E6 E10 e 7 11 e E8 El2 Install Analyzer HV DAC card in far left slot of the lower Bertan card cage with the component side to the right and the IC part of the card fac
231. Qu 90LS PMT1 LOG OUT IN PROGRAMMABLE J6 DCRESTORE GAIN AMPLIFIER 9018 OUT DECODE COMMANDS PN 4237003C FROM BACKPLANE CPU AND CONTROL CARD FUNCTIONS P1 BACKPLANE CONNECTOR 7003037A 2 4141 2 INSTRUMENT DESCRIPTION CARD DESCRIPTION NON GATED AMP NON SWITCH CONFIGURATION EXCEPT ANALYZER FL1 IN J3 FL2 IN J5 FL3 IN 3 PMT Sub Amp Card Function The 3 PMT Sub Amp card processes the FL3 signal to provide variable gain peak and integral signals and a log output See Figure 2 11 2 Jumpers In Figure 2 11 2 the jumpers set the bandwidth for sense in air or quartz flow cells systems and cards are set to the quartz configuration as the default For quartz operation Elite 76 u and 100 u sort sense flow cells X2 X4 X5 X6 X7 X8 X9 8 9 IN For sense in air operation X1 X2 X4 X5 X6 X7 X8 X9 X10 E8 E9 OUT Inputs The 3 PMT Sub Amp card accepts three FL inputs FL1 FL2 and FL3 Each input can have a percentage of one or both of the other inputs subtracted from it After subtraction the system outputs the FL1 and FL2 signals Figure 2 11 2 Block Diagram 3 PMT Sub Amp Card M 0 Note All divider outputs J2 Programmed Divider are inverted FL1 OUT Outputs to be subtracted Programmed Divider Outputs to be subtracted a SZ Programmed Divider J4 Outp
232. R BY CATEGORY Table 8 1 13 Lasers Continued Part Number Description Figure Item 7232506 Enterprise laser power supply air filter 1232507 Enterprise laser Temperature Sensor card 7232508 Enterprise laser hex wrench 7232509 Enterprise laser washer screen fine 7232510 Enterprise laser starter module 7232511 Enterprise laser water flow transducer 7232512 Enterprise laser head pc card 7232513 Enterprise laser beam splitter assembly 7232514 Enterprise laser light pick off 488 nm 7232515 Enterprise laser light pick off UV 7232516 Enterprise laser cable remote module 7232517 Enterprise laser cable heat exchanger interlock 25 feet 7232519 Enterprise laser cable umbilical 1232520 Enterprise laser model 621 operator s manual 7232521 Enterprise laser cooling fan power supply 1232522 Enterprise laser washer hose coarse screen 7232523 Enterprise laser power cord 8 gauge Table 8 1 14 Miscellaneous Part Number Description Figure Item 2016298 Drive floppy 5 25 in 8 2 28 4 2016328 Drive floppy 3 5 in 8 2 28 5 2121023 Jumper discrete wire 2 position single row 2523451 O ring sheath tank cap silicone 2838068 Clamps break apart metal 2851763 Screw machine M6 x 12 mm 2851920 Screw machine metric M6 x 20 mm 3814206 Rail mounting beam translator micro optical 3814224 Rail table laser optical 16 5 in Gloves latex 5415179 small medium 5415174 medium large 5415175 large
233. RAMMABLE J2 GAIN AMPLIFIER FL1 PEAK OUT 3 INTEGRATOR 1 PROGRAMMABLE e GAIN AMPLIFIER peeesesn FL1 LIN OUT A EN I pu py LOG AMP J4 FL1 LOG OUT PROGRAMMABLE J6 GAIN AMPLIFIER FL2 PEAK OUT gt INTEGRATOR e PROGRAMMABLE GAIN AMPLIFIER J7 FL2 LIN OUT HN ot t1 106 J8 ji FL2 NE LOG OUT A bk A I I SET A INTEGRATOR mI SPEED DECODE COMMANDS ji SET AMPLIFIER BANDWIDTH FROM BACKPLANE CPU IL GAINS emi ANDCONTROL 5 3 CARD FUNCTIONS P1 BACKPLANE CONNECTOR Scat CV SW R Card The Scat CV SW R card accepts FALS and 90LS inputs The FALS signal passes through a variable attenuator FALS HV control and is integrated The card provides the FALS signal in parallel to a variable gain linear amplifier and a log amp See Figure 2 10 3 7003057A This card also incorporates two multiplexer circuits The first Mux circuit selects either of the FALS signals as an additional output J4 The second Mux used only in a Gated Amp system selects one of the four Mux inputs to go to J9 2 10 2 PN 4237003C PN 42370036 INSTRUMENT DESCRIPTION CARD DESCRIPTION ALL CONFIGURATIONS Figure 2 10 3 Scat CV SW R Card Block Diagram
234. RATIONS 4 10 6 Table 4 10 6 Cable Connections Continued From To Quad PSH 2 J2 ADC J5 Quad PSH 2 J4 ADC J6 Quad PSH 2 J6 ADC J7 Pulse Pileup Det TOF J6 ADC J8 ADC 7 in cable J9 ADC J17 Sensor Interface J1 Pneu J4 Sensor Interface J2 Pneu J3 Sensor Interface J3 Multibus CPU J1 Sensor Interface J4 Multibus CPU J2 Non Gated Amp Units Scat CV J10 Peak Scatter Mux J12 Peak Scatter Mux J14 Mux and Scope J9 Scat CV J4 Mux and Scope J10 Scat CV J3 Mux and Scope J11 Scat CV J2 Mux and ScopE J12 Dual FL J3 Mux and Scope J13 Dual FL J4 Mux and Scope J14 Dual FL J2 Mux and Scope J15 Dual FL J7 Mux and Scope J16 Dual FL J8 Mux and Scope J17 Dual FL J6 Mux and Scope J18 3 PMT Sub J8 Mux and Scope J19 3 PMT Sub J7 Mux and Scope J20 3 PMT Sub J6 Mux and Scope J21 3 PMT Sub J4 Dual FL J1 3 PMT Sub J5 Dual FL J5 Gated Amp Units 7 Microsecond Delay 2 slot 8 J2 20 40 60 Microsecond Delay 2 slot 7 J2 7 Microsecond Delay 2 slot 8 J4 20 40 60 Microsecond Delay 2 slot 7 J5 7 Microsecond Delay 2 slot 8 J6 20 40 60 Microsecond Delay 2 slot 7 J8 PN 4237003C PN 4237003C ELECTRONIC ADJUSTMENTS CABLE AND JUMPER CONFIGURATIONS Table 4 10 6 Cable Connections Continued SERVICE AND REPAIR PROCEDURES From To 7 Microsecond Delay 2 slot 8 J8 20 40 60 Microsecond Delay 2 slot 7 J11
235. RIPTION ACQUISITION SUBSYSTEM 90800 peresueduioo p c ANS LINd LNI SLINd 30193 09 334 XV02 1 29 dV 14 z INd L 90 LNd A9 1V9S INI ST 039 3909 pue YOSNAS YALLVOS WALSAS S 1925 sdury LINdS 0L 9 2 91614 42370036 2 6 14 INSTRUMENT DESCRIPTION 2 ACQUISITION SUBSYSTEM 5PMTs Switchable Amps No Gated Amp The 5PMT Switchable Amps Figure 2 6 11 adds a fifth PMT and PMT Sub SW R card With this setup FLI and FL2 are routed to the Dual FL SW R card for linear and log amplification e The 3 PMT Sub SW R card completes processing the FL3 signal e Coax cables take all processed signals to the Mux and Scope card This configuration uses the Peak Scatter FALS sensor e Coax cables carry the FALS signal to the Peak Scatter Mux SW R card The Peak Scatter Mux SW R card splits the signal in two One signal is routed to J12 and then to the Scat CV SW R card which processes the signal to provide the integral and log FALS signals The second signal leaves the Peak Scatter Mux SW R card from J14 where coax cables carry the signal directly to the Mux and Scope card PN 4237003C 2 6 15 os EE amp Go gt SES gt eo ke 25 SE 22 E S lt 990001 8 981 LINd SY
236. Remove the scatter sensor Remove the laser shutter Remove the beam shaper Raise the flow cell all the way using the flow cell vertical adjustment knob 9o N uod Install the two smaller laser alignment targets on the vertical optical plate a Install the small targets b Install the first dichroic c Align the first laser through the small target holes d Repeat steps a through c for additional lasers 9 Atthe Cytometers Control Align screen turn on the HeNe and air cooled Argon lasers 10 Turn on the UV laser using the switch located on the laser 11 Close the output shutters on the air cooled Argon and the UV lasers to prevent beam over exposure Note At this point only the HeNe laser should be emitting a beam 12 Align the HeNe laser a Use the 90 degree dichroic mirror to align the HeNe through the two vertical optical plate mounted targets b Close the shutter on the HeNe laser 13 Move the two optical bench mounted targets to the front of the air cooled Argon laser 14 Open the shutter on the air cooled Argon laser 15 Align the Argon laser a Use the 90 degree dichroic mirror to align the Argon through the two vertical optical plate mounted targets b Close the shutter on the air cooled Argon laser WARNING Risk of personal injury due to ultra violet laser beam exposure Remove rings and other metal jewelry before continuing The laser beam can cause eye damage if viewed either directly or i
237. SIUL 100 wamo7 Lxnv GA 1 Wadan peg H jeddn wu uofuy sweaq 119 LXNY se 99 656 ZLINd ubissy 9 aq 0 9 7 ureeq 190 941 10 wgddn 1eddn y jeuBis 10 391049 uonouni al 7 L 93 E 318VSIq 318VN3 AY130 TVN9SIS se 10 aseaJoul NIV9 sseJd o u98J9S UIJ y uo pa1e20 0 09 NIV9 5x V3d 3 LNI INI LI Teuuou 94 0 1019 1 uonosjes NIV9 dI AV3ud SIUL uybuaJjs jeufiis jnjasn alow e Ajgissog paseaJour dn Aq jeuDis Dot 1 y pue jou Dot s Jaysew au Hurda y 192 aures y uo 1 pue uy Buipeap njdjau 1 al Bunos LINd 1ejnonued y uey euis 1942316 0 dn xv ay swojpe SL OL 0 DL Woy paysnipe NIV ASAPOV NIV9 dNYIYd ZXNY ANY Exnv AINO SLIM dA V 03159 LINd 304 x anon 7 eid 0L 01 NIV9 NIV9 dd
238. Sample Pressure Systems 2 9 4 Old Type 2 9 4 New 2 9 7 Sheath Pressure Systems 2 9 8 Old Type 2 9 8 New Type 2 9 10 Compressor Module 2 9 12 CARD DESCRIPTION ALL CONFIGURATIONS 2 10 1 3 PMT Sub SW R Card 2 10 1 Dual FL SW R Card 2 10 1 Scat CV SW R Card 2 10 2 Peak Scatter Mux SW R Card 2 10 3 CARD DESCRIPTION NON GATED AMP NON SWITCH CONFIGURATION EXCEPT ANALYZER 2 11 1 Scat CV Amp Card 2 11 1 3 PMT Sub Amp Card 2 11 2 Function 2 11 2 Jumpers 2 11 2 Inputs 2 11 2 Dual FL Amp Card 2 11 3 Function 2 11 3 Inputs 2 11 3 CARD DESCRIPTION GATED AMPLIFIER CONFIGURATION 2 12 1 Gated Amp Control Card 2 12 1 Gated Amp R3 Card 2 12 2 Functions 2 12 2 Scat Aux Gated Amp Card 2 12 2 Functions 2 12 2 Inputs 2 12 2 PMT Gated Amp Card 2 12 4 Function 2 12 4 Inputs 2 12 4 20 40 60 Microsecond Delay Card 2 12 6 PN 4237003C PN 42370036 7 Microsecond Delay Card 2 12 6 2 13 SIGNAL SELECTION AND DIGITIZATION 2 13 1 Overview 2 13 1 Signal Flow Description 2 13 2 Signal Source 2 13 2 Mux and Scope Card 2 13 2 Quad Peak Sense and Hold PSH Cards 2 2 13 3 ADC and PSH Control Card 2 13 5 ADC and PSH Timing 2 13 8 Peak ADC Card 2 13 9 ADC and PSH Control Card 2 13 11 Lister Out Card 2 13 12 Lister A B Card 2 13 14 3 INSTALLATION PROCEDURES 3 1 1 3 1 3 2 3 3 et 32 PART A INSTRUMENT INSTALLATION PREINSTALLATION 3 1 1
239. Sensor Interface card 2 9 4 acquisition check creating a protocol 3 6 1 acquisition start delay See ASD Acquisition subsystem analog signal processing description 2 6 1 description 2 6 1 PMT configurations 2 6 1 troubleshooting how to 7 2 3 ADC and PSH Control card block diagram 2 13 6 function 2 13 5 2 13 11 ratios calculated 2 13 5 ADC card function 2 13 8 ADC timing illustration 2 13 12 adjustments Autoclone Sorting Option mechanism 2 2 4 Autoclone Sorting Option speed 3 10 7 Autoclone Sorting Option stream 3 10 9 Autoclone Sorting Option waste catcher 3 10 9 Autoclone Sorting Option See Autoclone Sorting Option camera image R99 of Camera Interface card 4 1 11 camera See camera electronic pressure regulator 4 6 1 low bleed regulator 4 5 1 non low bleed regulator 4 5 4 optical knobs for 4 12 2 Pneumatic Interface card See Pneumatic Interface card porch settings front and back 3 6 5 pressure switch 4 7 1 sample regulator 4 5 3 sheath regulator 4 5 3 sort pulse amplitude 4 2 3 air water separator See Compressor module alignment procedures INDEX 1 INDEX Argon laser 3 5 4 beam translator equipped instruments 4 1 3 collinear laser alignment procedure 3 12 4 flow cell 4 1 3 4 12 1 flow cell after Piezo bimorph installation 3 16 2 HeNe laser 3 5 4 laser beam 4 12 1 lasers for Gated Amp instruments 4 1 4 parabolic mirror on tip optional 4 12 4 standard laser 3 12 1 4 1 1 t
240. Service Bulletin 1328 Changes were made on the following pages 1 2 2 1 2 9 2 3 13 2 5 1 2 5 2 2 8 4 2 9 1 3 1 1 3 1 3 3 9 1 3 13 1 through 3 13 5 3 22 1 through 3 22 2 4 10 1 through 4 10 2 6 1 1 through 6 1 4 8 1 1 8 1 2 8 1 3 8 1 6 8 1 7 8 1 16 8 1 18 8 1 20 8 1 22 8 2 5 8 2 8 8 2 9 8 2 11 8 2 12 8 238 A 2 2 ABBREVIATIONS TRADEMARKS Additionally Installation procedure 3 22 CYTOMETER CPU CARD UPGRADE was replaced with ELITE 256 SORT UPGRADE Heading 4 13 EXTERNAL MEMORY CARD REPLACEMENT was added e Heading 7 6 RESETTING CYTOMETER SOFTWARE LOCKUPS was added Under Heading 8 1 MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY Table 6 Cytometer and Table 7 Cytometer were combined As a result table numbers 8 and above are decreased by one For example Table 8 1 8 is now Table 8 1 7 and so forth Since check digits are not used in the VANTIVE or CARES systems all check digits have been removed from the part numbers in Chapter 8 To avoid confusion when ordering parts that entire chapter has been added to the change page packet Changes that are part of the most recent revision are indicated in the printed copy by a bar in the margin of the amended page This document applies to the latest software listed and higher versions When a subsequent software version affects the information in this document the changes will be included on minor revision change pages or summarized on a Notice of In
241. TB9 2B jumper brass TB9 3B to TB9 4B jumper brass TB9 5B to TB9 6B jumper brass 240 Vac input to main system transformer TB9 3B to TB9 5B jumper cable TB9 1B to TB9 2B jumper brass TB9 4B to TB9 8B jumper cable TB9 6B to TB9 10B jumper cable 100 Vac input to laser auto transformer TB10 1B to TB10 4B jumper cable TB10 2B to TB10 8B jumper cable TB10 6B to TB10 9B jumper cable TB10 8B to TB10 10B jumper cable TB10 4B to TB10 5B jumper brass TB10 6B to TB10 7B jumper brass Remove TB10 1B to TB10 10B jumper cable 115 Vac input to laser auto transformer TB10 1B to TB10 10B jumper cable 230 Vac input to laser auto transformer TB10 1B to TB10 4B jumper cable TB10 6B to TB10 9B jumper cable TB10 8B to TB10 10B jumper cable TB10 7B to TB10 8B jumper brass Remove TB10 1B to TB10 10B jumper cable 240 Vac input to laser auto transformer TB10 1B to TB10 3B jumper cable TB10 6B to TB10 9B jumper cable TB10 8B to TB10 10B jumper cable TB10 7B to TB10 8B Jumper brass Remove TB10 1B to TB10 10B jumper cable 4 3 1 SERVICE AND REPAIR PROCEDURES LINE VOLTAGE SELECTION 4 3 2 PN 4237003C SERVICE AND REPAIR PROCEDURES POWER SUPPLIES MEASUREMENT AND ADJUSTMENT 4 4 POWER SUPPLIES MEASUREMENT AND ADJUSTMENT Purpose Perform this procedure after the Pneumatic Interface card regulator sensor or t
242. TEM Purpose Do this procedure to troubleshoot the Acquisition subsystem Tools Supplies Needed None Procedure 1 Use Figure 7 2 1 Acquisition Troubleshooting Flowchart as a guide when troubleshooting the Acquisition subsystem 2 Ifthe problem persists see Acquisition Troubleshooting Methods for additional instructions 4237003 7 2 1 TROUBLESHOOTING TROUBLESHOOTING THE ACQUISITION SUBSYSTEM Figure 7 2 1 Acquisition Troubleshooting Flowchart ACQUISITION Troubleshooting Guide Create protocol to acquire Test pulses Can FI Test pulses be acquired Yes Y Can Fluorespheres beads No Yes be acquired v v v SAMPLE OPERATION Optical Fluidics Electronics acquires beads problems with cells cannot acquire beads Use CYTO TROL control cells to test If OK consider 3 aser s problem cosider what Use protocol as a tool Laser Alignment the instrument must do Simplify protocol until system acquires UR the then add features until problem per aiig ensure the following are Refer to Optical Alignment Reorder parameters to reroute signals Procedures e light filters elaser s e beamshaper eflow Cell discriminators i No eret Does sheath flow out of lower beam lockout flow cell tip Yes ET Remove sample tube and go to Run Does sheath drip from sam
243. TENTION The paper size to be used must not contain a REM statement before it Press Ctri C L and press Enter Verify that the 1 DRIVI DRIVI DRIVI CGI CFG file matches the following ER C ELITE GSS GSSCGI SYS ER C ELITE GSS META SYS G OUTPUT ER C ELITE GSS HRVGA SYS G DISPLAY DISPLAY HRVGA SYS ELITE GSS MSMOUSE SYS G INPUT MSMOUSE SYS Lj 2 3 4 5 DRIVI 6 7 8 MOUSI MOUSI 9 10 rem 11 rem 12 rem 134 14 rem 15 rem 16 rem 17 18 rem 19 rem 20 rem 24s 22 rem 23 rem 24 rem 25 rem 26 rem FONTS C ELITE GSS standard laser printer driver DRIVER C ELITE GSS LASERJET SYS G PRINTER RESOLUTION 75 standard dot matrix printer driver DRIVER C ELITE GSS IBMGPR SYS G PRINTER RESOLUTION 75 HP Paintjet driver 90 dot per inch printer driver DRIVER C ELITE GSS HPPJ90 SYS G PRINTER Easy 2 users printer driver configuration DRIVER C ELITE GSS EPSONX SYS G PRINTER PRINTER EPSONX SYS EPSONX PRN RESOLUTION 120 3 20 5 3 INSTALLATION PROCEDURES COLOR PRINTER UPGRADE 27 28 DRIVER C ELITE GSS HPPCL5 SYS G PRINTER 29 HPPCL5 PRN 30 RESOLUTION 300 31 COLORS 16 32 FF ON 33 3 34 ORIENTATION PORTRAIT 35 rem PAPER ISOA4 36 PAPER LETTER 37 PLISTSIZE 4096 38 rem F
244. TION ATTENTION Users of pre 486 computers will experience a reduction in system operating speed ATTENTION Country service organizations must track the installation of Software Version 4 0 in instruments with the Autoclone option The international implementation status must be reported through the field Corrective Action Status by Country form Purpose Use this procedure to install Elite Software Version 4 0 in all instruments currently with version 3 25 software Version 3 25 was installed when the Autoclone option was installed The Autoclone Version 3 25 serial numbers affected are 00N44015 00N44016 00509119 00513191 00535515 00835572 00539664 00539665 00544740 00544743 00552904 00707115 00708151 00130519 00133553 00146776 00746777 00746778 00005081 00008166 00008167 00012302 00018397 00023601 00023602 00032613 00036621 00036623 00040632 00049649 00049650 This procedure also applies to Elite Analyzer Flow Cytometer 2819 Elite Software single 2365 Elite Software multiple 2366 e High SN ORSO9119 Low SN V39054 Note For Elite Analyzers operating with version 3 0 or 3 1 software version 4 0 is considered an enhancement Version 4 0 software is required and should be installed on instruments that include the Panelyzer option Installation of additional options may require version 4 0 software Installation in stand alone workstations can be do
245. The Cytometer Control screen disappears during the software transfer Do Waste Bottle Modification for Software Version 4 01 When the Control screen appears select POWER UP VALVES and wait for the valve action to be completed 9 Ifthe instrument is equipped with an Autoclone Sorting Option do Calibration Procedure under Heading 3 10 AUTOCLONE SORTING OPTION INSTALLATION to allow for proper drainage in the Sort Collection area Note Even if the Autoclone Sorting Option is not being used at this time you must still do the Calibration Procedure Waste Bottle Modification for Software Version 4 01 The Elite uses a magnetically actuated switch to detect when the waste bottle is full In instruments with version 4 0 or earlier software the switch is closed when the bottle is full The software interprets this switch closure as Waste Bottle Full error condition If the switch fails or if the user fails to reconnect the switch after emptying the bottle the bottle full condition may not be detected Although the instrument incorporates other safeguards the software logic of version 4 01 detects an open switch condition This software change combined with the modification to the level switch provides a self monitored system This means that if the user fails to reconnect the switch or if the switch fails the Waste Bottle Full error message appears on the Cytometer Control screen ATTENTION Software Version 4 01 must be installed befor
246. There are various configurations for this part of the system e 4PMT Nonswitchable Amps without Peak Scatter No Gated Amp e 4 PMT Switchable Amp with Peak Scatter No Gated Amp e 5 PMT Nonswitchable Amps without Peak Scatter No Gated Amp e 5 PMT Switchable Amps with Peak Scatter Gated Amp For information on the signal flow for each configuration see Signal Flow for Each Configuration in this section All the configurations Develop integral signals e Provide programmed linear gain of peak and integral signals e Develop logarithmically amplified signals e Provide subtraction FL compensation Subtraction Color Compensation Color compensation allows the use of dyes that have overlapping emission spectra Electronic subtraction circuits are used to perform color compensation Figure 2 6 1 shows the operation of a single subtraction circuit e FL2 is divided by a resistor network to provide the desired subtraction percent The values shown in Figure 2 6 1 are for 2096 1 5 In the actual circuit an electronically controlled divider called a DAC is used The circuit is configured to add two voltages e The output is FL1 2096 FL2 4 PN 4237003C 2 6 1 2 INSTRUMENT DESCRIPTION ACQUISITION SUBSYSTEM Figure 2 6 1 Subtraction Circuit Example FL2 pulse passes through the circuit unchanged Iy 40 KQ This preamp inverts the pulse FL1 pulse ST 7003056 Multiple circuits pe
247. UBSYSTEM Figure 2 9 4 Block Diagram Sheath Pressure System Old Type ccu TP10 ge 10 00 V VOLT REF emm NPUTLATCH wm DAC 11 1 1 1 1 1 1 10 00 V m R54 FEEDBACK 1 RISGA vou AND R76 ZERO REF DRIVE 1 REGULATOR LIQUID PRESSURE TRANSDUCER 0 10 IN 499909900990929992 999 3 15 PSI OUT INSTRUMENT DESCRIPTION E 7003075 Regarding Figure 2 9 4 30 PSI IN The Input Latch is an 8 bit word set pressure written by CPU U17 The DAC provides a voltage proportional to the digital pressure Full scale is set by Volt Ref 2 9 9 2 INSTRUMENT DESCRIPTION PNEUMATICS SUBSYSTEM New Type With the new type of Sheath Pressure system Figure 2 9 5 the circuit is slightly more complex than the Sample Pressure Regulation circuit because the system measures sheath pressure using a liquid pressure transducer at the output of the sheath tank The amplified pressure transducer output goes to the Sensor Interface card for display on the Cytometer s Control screen In normal operation the amplified pressure transducer is connected to the pressure regulator to provide a feedback signal The Pneumatic Control card can also be put in a tes
248. V 435 Brown 424V None 24 24V 436 Grn Yel CHAS GND None AC NEUT None AC NEUT None AC HOT None AC HOT 34 If wires from the deflection body holder are running down in front of the optical plate a Taking care to note their location remove the wires from the terminal block assembly b Route the wires horizontally along the optical plate toward the back of the unit c Wrap the wires along the back and insert wires down through the hole on the PMT plate d Reconnect wires to terminal block assembly in their original location 3 10 5 INSTALLATION PROCEDURES AUTOCLONE SORTING OPTION INSTALLATION 3 10 6 35 36 3T 38 39 40 41 42 Reroute any other cables or tubing coming up along the front of the optical table through this same hole on the PMT plate Place a wire tie mount on the front of the optical plate taking care it does not interfere with anything Use a wire tie to clamp all the cables and tubing to the wire tie mount Replace the terminal block assembly and fasten with screws from step 31 Place the Autoclone Sorting Option mechanism under the optical table with 37 pin D connector to rear of Cytometer directly behind the sample collection area Check the level of the Autoclone Sorting Option to Cytometer optical table Ifthe Autoclone Sorting Option is not level use double sided tape to fasten the metal strip to the top of the inner frame rail
249. Vdc TP3 R51 8 00 0 01 Vdc TP2 4 5 1 4 SERVICE AND REPAIR PROCEDURES PNEUMATICS SYSTEM 10 11 12 13 14 15 16 1T 18 19 20 21 22 23 24 Remove the jumper from EI to E2 and place E2 to Connect the external pressure gauge to the sample pressure by plugging a T fitting into the pinch tubing at the sample pressure pinch valve far left fitting Set Sample Pressure to 11 5 psi at the Cytometer Control Valves screen Adjust R34 slowly until the external gauge reads 11 5 0 02 psi This should require no more than two turns of the potentiometer If more than two turns are required there may be a problem or your test equipment is not accurate enough for this procedure Correct the problem and repeat steps beginning at 1 Adjust R51 slowly until read pressure on Cytometer screen is 11 5 0 02 psi Set Sample Pressure to the following pressures and verify that the set read and gauge pressures agree within 0 1 psi 11 5 psi 4 5 psi Repeat steps 6 through 8 if needed Set Sample Pressure to 5 00 psi Verify that the set read and gauge pressures agree within 0 2 psi Repeat steps 6 through 8 if needed Remove the external gauge from the sample pressure line and tee in the sheath pressure at the sheath pressure pinch valve middle fitting Set Sheath Pressure to 15 00 psi on the Cytometer screen Adjust R81 for 15 00 0 02 psi on the gauge Adjust R75 for 10 00 0 01 Vdc a
250. W SLOT 12 DELAY DELAY 3 PMT SUB SWITCH R 3 E1 E2 IN E3 E4 OUT FL1 QUT E5 E6 IN y 1 QUAD 20 40 60 SLOT 10 LEGEND FOR E1 E2 IN COAX USED 24 EACH USE 27 COAX CABLE 8027803 USE 17 COAX CABLE TESTED BOARD 51515 PART NUMBERS SUB SWITCH 2 QUAD 20 40 60 6705246 81818 SLOT 7 SCATICV SWITCH amp i i 6705252 01010 E1 E2 IN PEAK SCAT MUX SW 4 IN E10 E11 6705389 e DUAL FL SWITCH to NEN 6705249 5 HI PEAK SCAT SENSOR ii 6705182 E leie GATED S555 2055 S555 6704008 TO FROM SLOW SCAT AUX GATED a DELAY DELAYS DELAY 6704009 J3 QUAD 20 40 60 AUX2 OUT Ji 6704904 TRIG SCR OUT io QUAD 7 DELAY AUX1 OUT 6704241 J4 PMT 2 mE PMT 2 IN PMT 3 IN d PMT 4 Kr EE PMT 5 PRX PMT 5IN PMT GATED kee E SLOT46 og 3 1 2 007 6 OUT _________ _ SLOT 1 SCATTER MUX SWITCH R 1 AUX 1 IN AUX 2 INT IN AUX 2 LOG IN AUX 2 PK IN PMT2 PK E1 E2 IN E1 E2 IN PARAM B FALS PK FALS PK AUX 1 INT AUX 1 LOG FALS IN SLOT 5 3PMT SUB SWITCH R 1 E1 E2 OUT 4 OUT E5 E6 OUT SLOT 4 SUB FL 3 INT
251. a Adjustment Procedure 4 1 9 Aiming the Camera 4 1 9 Focusing the Camera 4 1 10 Zooming the Camera 4 1 10 Positioning Movement of the Camera 4 1 11 Optimizing the Image 4 1 11 42 SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE 4 2 1 Purpose 4 2 1 Tools Supplies Needed 4 2 2 Crystal Drive Test Procedure 4 2 2 Crystal Drive Adjustment Procedure 4 2 2 Sort Pulse Amplitude Test 4 2 2 Sort Pulse Amplitude Adjustment 4 2 3 Phase Settings 4 2 3 Delay Settings 4 2 3 Sync Adjustment 4 2 4 Sort Function Verification 4 2 4 Drive Frequency Sort Settings 4 2 4 Phase Adjustment 4 2 5 Delay Calculation 4 2 7 Monitoring the Sort 4 2 8 Sort Coincidence Verification 4 2 9 Tools Supplies Needed 4 2 10 Setup Pulse Generator 4 2 10 Acquire and Setup Sort 4 2 10 Pulse Pileup Verification 4 2 11 Verification of Sort Pulse Extension 4 2 12 Sort Pulse Truncation Test 4 2 12 Extended Abort Test 4 2 13 PN 4237003 4 i 4 1 4 3 4 4 45 4 6 4 7 4 8 4 9 4 10 Pulse Generator Card Adjustment 4 2 14 LINE VOLTAGE SELECTION 4 3 1 POWER SUPPLIES MEASUREMENT AND ADJUSTMENT 4 4 1 Purpose 4 4 1 Tools Supplies Needed 4 4 1 Procedure 4 4 2 PNEUMATICS SYSTEM 4 5 1 Low Bleed Regulator Adjustment Procedure 4 5 1 Purpose 4 5 1 Tools Supplies Needed 4 5 1 Pneumatic Interface Card Adjustment 4 5 1 Low Bleed Regulator Adjustment 4 5 2 Non Low Bleed Regulator Adjustment Procedure
252. a stepper motor and has optical sensors to determine when the stage is in the full forward load position back to the end of the tray home position and the full backward retracted position The stage also carries overtravel switches to give an error condition if the stage moves too far forward or back Optical Sensors You can see the status of the optical sensors on the Autoclone Sorting Option screen on the Cytometer in the box labeled Markers Only those sensors currently read appear on the screen See Table 2 2 1 PN 4237003C INSTRUMENT DESCRIPTION AUTOCLONE SORTING OPTION Table 2 2 1 Autoclone Sorting Option Screen Marker Definitions Markers on Screen Description Lx Tray is in the full forward load position Row 12 of a 96 well plate is under the flow cell when this marker is displayed Hx Tray is in the back home position Row 1 of a 96 well plate is under the flow cell when this marker is displayed Ly Tray is all the way to the left Column A or column H of a 96 well plate is under the flow cell when this marker is displayed Hy Tray is all the way to the right Column D or column E is under the flow cell when this marker is displayed Cw Tray is in the load rotational position the cutout is to the front CC Tray is rotated 180 degrees the cutout is in the back Hx and Lx both appear Arm is in the fully back retracted position CC and CW both appear ERROR condition tray holde
253. adjustments to keep the Elite operating smoothly This schedule is only a recommendation and may vary with instrument usage and environment Tools Supplies Needed Elite PMI Kit PN 6913419 0 Brass T fittings PN 6216129 9 Tubing connecting the T fittings PN 3202039 9 Waste carrying tubing PN 3213194 8 PN 3202039 9 on older systems Hydrophobic filter round disk type 0 3 p PN 6232143 1 Halogen lamp PN 3908025 7 Metal holder clamps for both sheath and waste filters PN 2838068 2 PN 3203027 1 Sheath filter interconnections PN 6232472 4 PN 6232475 9 PN 6232522 4 Sheath filter PN 6232473 2 Waste filter PN 6232489 9 Vial cap PN 6856967 2 Check valves in the waste lines PN 6214106 9 PN 6801536 7 Male and female waste tank connectors PN 6232429 5 PN 6232452 0 PN 6232546 1 PN 6232547 0 Sheath tank cap O ring PN 2523451 1 Before You Begin l Inspect the instrument for operational problems and correct if found 2 Do the appropriate portion s of the VIP under Heading 5 1 VERIFICATION INSPECTION PROCEDURE to verify operation of the repaired component s Procedure 1 Ensure that the kit is complete by comparing the contents with the packing list Turn the unit OFE Open pneumatics covers Replace all the brass T fittings Replace all tubing that connects the T fittings Replace all remaining waste carrying tubing Mma Me WY Replace hydrophobic filter r
254. agram PMT Gated Amp Card FAST 7 MICROSECONDS SLOW 20 40 60 MICROSECONDS DELAY CARD 2 DELAY CARD 2 This is a quad This is a quad 4 channel card 4 channel card J8 TO DELAY J12 CARDS J13 This analog switch choses between fast and slow delay Note that all channels are set the same S This switch chooses delays undelayed These are individually controllable J4 PMT 2 DC e F IN ide 1 9 me SENSOR 1 OUT BS gi ke 1 5 2 THIS SWITCH ALLOWS Ge THE SIGNAL TO PASS 49 J14 J15 OR GROUNDS THE Bag SENSOR 2 OUT OUTPUT e PMT 3 l a TET Ale SENSOR 3 OUT J5 Ii ev PMT4 PMT3 DC RESTORE e He gp cre IN E 1 l Ka jr id 0 TRIGGER BEEN MCN bo multiplexed where o SSS 554 SETBYI O These commands originate GATEPULSE each output COMMAND FROM at Gated Amp control where from PROGAMMABLE CPU they are decoded from GatedAmp of the bd P2 ATTENUATOR
255. al Autoclone Sorting Option function of 2 2 2 2 2 3 Serial I O card function 2 5 2 jumpers 4 10 1 A 2 2 serial numbers version 3 25 software instruments with 3 18 1 set pressure See pneumatics settings delay calculation procedure 3 6 5 drive frequency procedure 3 6 3 droplet delay calculating 4 2 7 phase adjustment procedure 3 6 3 setup camera initial 4 2 7 setup instrument See instrument setup sheath leaks air testing for 7 3 2 leaks liquid testing for 7 3 2 sheath pressure troubleshooting problems 7 4 1 sheath pressure system new type block diagram 2 9 11 description 2 9 10 sheath pressure system old type block diagram 2 9 9 description 2 9 8 sheath pressure valve VL6 function of 2 9 2 sheath regulator adjustment See pneumatics short pass SP filter function 2 3 12 42370036 signal flow description 2 13 2 source of signals 2 13 2 summary of various configurations 2 6 8 signal selection and digitization overview 2 13 1 Snell s Law equation 2 7 2 software Autoclone Sorting Option 2 2 4 Elite 2 1 2 Elite software installation of 4 9 1 files location of B 1 1 for the Workstation 4 8 1 HP 1600C color printer software installation 3 20 2 installation general 4 9 1 PC Model 486 See PC Model 486 version 4 0 installation 3 18 1 version 4 0 when required 3 18 1 version 4 01 installation 3 19 1 waste bottle modification for software version 4 01 3 19 2 3 19 3
256. al Troubleshooting Information 7 1 2 General Troubleshooting Information 7 1 3 Lasers General Troubleshooting Information 7 1 5 Optics General Troubleshooting Information 7 1 5 Electronics General Troubleshooting Information 7 1 6 Software General Troubleshooting Information 7 1 6 Power Supplies General Troubleshooting Information 7 1 6 Fluidic Components 7 3 1 Troubleshooting Sort Problems 7 5 2 PN 4237003C 7 1 ISOLATING THE PROBLEM AREA Purpose This is a general procedure for isolating a problem area The troubleshooting charts and TROUBLESHOOTING methods in this section are examples of recommended troubleshooting approaches for use with common problem areas Refer to the Special Procedures and Troubleshooting manual for further information Tools Supplies Needed None Procedure l Dostartups alignment check or other symptom checks to obtain a clear symptom 2 Use the symptom to isolate the problem area 3 Check the problem area for the types of problems shown in Table 7 1 1 Table 7 1 1 Isolating Problem Areas Problem Area s Possible Causes Biological sample Bad reagents Poor preparation Abnormal sample Electronics Adjustment Poor connection Defective components Filters Wrong filter Dirty filter Defective filter Fluidics sheath sample or waste Pinch Plug Leak Defective component Laser s No beam No power Current too high
257. al alignment principle adjust the front laser mount to align to the target closest to the laser Adjust the rear laser mount to align to the target furthest from the laser series of small iterative adjustments works best PN 4237003C PN 42370036 SERVICE AND REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE Flow Cell Alignment Initial Use this procedure to center the flow cell channel in the laser beam The flow cell face must be normal perpendicular to the laser beam Remove the beam shaper 2 Verify that the laser beam is centered on the scatter sensor If not or if there is any doubt as to laser alignment install the targets align the laser 3 Turn Sheath on and open the laser shutter Adjust the flow cell vertically to ensure the laser beam is passing through the quartz cell 5 Rotate the flow cell so the laser beam reflects back through the laser a Loosen the nut securing the flow cell to the bimorph b Rotate the flow cell as needed c Tighten the nut 6 Adjust the flow cell tilt so the laser beam reflects back through the laser shutter a Loosen the two bolts securing the flow cell stage to the vertical plate only enough to allow the flow cell to be tilted b Adjust the flow cell tilt as needed c Tighten the bolts 7 For systems with the flat bimorph Slightly loosen the two bolts which secure the bimorph plates to the flow cell stage b Slide the plate and flow cell assembly in order to center the
258. al rail to mount a single laser target Verify that the a Translator knobs faces away from the laser b Translator window is perpendicular to the optical bench Adjust the translator height in post for the laser to penetrate as close as possible to the center of the translator window Refer to the multi hole target and set the alignment mode for lowest target hole Move the pin on the bottom of the target to line up with the desired target hole and lock down the pin with setscrew Install the lowest target in the space between the beam translator and the Argon dichroic and ensure that the target is square with the table top Rotate the translator window so that the Argon beam is aligned with the lowest laser target hole Remove the lowest Argon laser target Use the adjustable Argon dichroic on the optical rail look at the lower trace on the Digiscope Screen and adjust the trace between the peak of the Argon caused signal and the peak of the UV caused signal The adjustment should be e 20 microseconds for a 100 pm quartz tip 30 microseconds for a 76 um quartz tip 3 5 microseconds for a 76 pm jet in air tip Note The UV signal is the middle one on the Digiscope Test this by increasing and decreasing the UV laser power and observing the far middle peak that follows At this point the beams should be Argon UV and HeNe Refer to the multi hole target and set the Alignment mode for highest hole Move the
259. aller forms this population G Both PMT 2 peaks are acquired as one population G COUNT COUNT 0 0 T T T T T 1 T1 71 1 T 1 T T 0 200 400 600 800 1000 0 200 400 00 800 1000 2 2 7003179 Pulse Generator Card Adjustment 1 Put the Pulse Generator card on the extender and ensure jumpers X1 X2 X4 are in place Install jumpers if necessary Attach the scope to 1 Use the Service screen to turn on double pulses a At the Cytometers Service screen access code 26 28 32 select Debug 2 b Ifthe Mode box does not display single press it until it does c Press the following blocks on the touchscreen PORT 0 2 0 2 OUTPUT D 6 0 1 START PORT 0 2 0 4 OUTPUT F F F F START 4 Adjust the card for the values in Table 4 2 1 4 2 14 PN 4237003C SERVICE AND REPAIR PROCEDURES SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE Table 4 2 1 Pulse Generator Card Values POT Function Test Point TP Adjust For R109 Delay TP1 100 microsecond delay between the two pulses R108 Rate TP1 1 millisecond delay between pulse pairs R54 Phase1 width TP1 12 microsecond R53 Phase2 width TP1 12 microsecond initial value R44 Ref volt TP19 10 V R75 Gain TP20 5 5 V pulse R107 Gain TP4 Maximum undistorted pulse
260. an open collector gate that permits all of these outputs to share a common line to the ADC and PSH Control card This common line is called DISCSAT and is normally pulled high to 5 V through a resistor Therefore when any discriminator on any PSH channel is exceeded the DISCSAT line is pulled low This signals the ADC and PSH Control card that a cell is present and that data needs to be acquired Captures and holds the highest voltage value of the incoming pulse The Peak Sense and Hold circuit performs this function The signal pulse can charge a capacitor through a diode This ensures that the capacitor is charged to the highest peak value of the incoming pulse and prevents the capacitor from discharging when the pulse ends The capacitor holds the charge until the ADC and PSH Control card sends a discharge signal back to the capacitor The pulse that leaves the card is the stretched pulse The PSH circuits input contains switch to ground that the ADC and PSH Control card controls This isolates the card s input from noise after a valid pulse is captured PN 4237003C INSTRUMENT DESCRIPTION SIGNAL SELECTION AND DIGITIZATION Figure 2 13 2 Block Diagram PSH Card s This is the Hold circuit The capacitor is charged to the peak value of the pulse The diode prevents the capacitor from discharging Signal is delayed for 1 microsecond at the end of the pulse The
261. and solenoids do not function properly See Table 7 3 1 for fluidic components Use Figure 7 3 1 as a guide for general fluidic troubleshooting guidelines Table 7 3 1 Fluidic Components Pilot actuator Air pressure moves a piston inside the actuator which then moves something else such as a pinch valve 7003184A Check valve Permits flow in one direction only 70031834 Pinch valve Controls the path of fluid flow 7003182A Solenoid Controls the pressurization of other components 7003181A PN 4237003 7 3 1 TROUBLESHOOTING TROUBLESHOOTING FLUIDIC PROBLEMS 7 3 2 Tools Supplies Needed Pressure and Vacuum gauge accurate to 01 psi range 0 to 30psi Hemostats Oh DVM Air Leaks Sheath Air Leak Testing 1 At the Cytometer select Powerup Valves and wait for sheath pressure to stabilize Note At this point the air in the sheath tank is pressurized to the Sheath pressure setting Press VACUUM and listen to the sheath regulator Ifyou detect a leak go to step 3 Note Since no liquid is leaving the Sheath Tank no additional air should be entering the tank If you hear a buzzing sound coming from the regulator this means air is being supplied to compensate for a leak Ifyou do not detect a leak go to step 4 Apply soapy water to the bottle top and connections to locate the source of the air leak If you cannot find the leak a Tee your pressure gauge to the regulator
262. and the center on the other side Front Porch is not set correctly With Sort Test on and 3 drops sorted adjust the Front Porch while watching the side streams until they form a single side stream Note Default setting is 8 Flow cell is suspect Replace the flow cell 7 5 6 42370036 Table 7 5 1 Troubleshooting Sort Problems Continued TROUBLESHOOTING TROUBLESHOOTING THE SORT SUBSYSTEM Condition Symptom Problem Corrective Action Cannot gettight There is no phase The stage housing for the Remove the Newport label on the bimorph stage side streams setting that enclosed bimorph may not be and tighten the two screws that preload the produces a clean secured tight enough which bearings in the stage housing Do this carefully side stream causes a slight vibration in to remove any free movement and to avoid the bimorph assembly binding movement of the housing Wide or fanning center stream Center stream is fuzzy not a clean single stream Back porch is not set correctly With Sort Test on and 3 drops sorted adjust the Back Porch while watching the center stream until it forms a single side stream Default setting is 2 Cannot get There is no Hydraulic or pneumatics leak Check the sheath tank cap sheath filter sheath sharp focused frequency setting or problem tubing through the system Use soapy water to drops that produces find the leaks when bubbles
263. ard signal flow function of 2 6 8 3 PMT Sub Amp 2 card signal flow summary of 2 6 8 3 PMT Sub Amp card block diagram 2 11 2 FL inputs accepted 2 11 2 function 2 11 2 3 PMT Sub Amp SW R card jumpers 4 10 3 A 2 2 A 2 3 PMT Sub Amp SW RI card jumpers 4 10 3 A 2 3 3 PMT Sub Amp SW R2 card jumpers 4 10 3 A 2 3 3 PMT Sub SW R card block diagram 2 10 1 function 2 10 1 jumpers for Gated Amp instruments 3 9 3 3 9 4 3 PMT Sub SW R2 card jumpers A 2 1 4 PMTs Nonswitchable Amp configuration block diagram 2 6 10 4 PMTs switchable amp with peak scatter function block diagram 2 6 12 5 PMTs Nonswitchable Amp No Gated Amp block diagram 2 6 14 5 PMTs switchable amp no gated amp block diagram 2 6 16 7 Microsecond Delay card block diagram 2 12 6 15 V 0 25A PMT power supply adjustments and measurements 4 4 2 A 3 2 15 V 3 4A power supply adjustments and measurements 4 4 2 A 3 2 15 V 5 0A power supply adjustments and measurements 4 4 2 A 3 2 20 40 60 Microsecond Delay card block diagram 2 12 6 function 2 12 6 90 V power supply adjustments and measurements 4 4 2 A 3 2 9096 bead requirement for sorting 3 6 6 256K Memory card batteries Lithium life of 2 5 2 function 2 5 2 jumpers 4 10 2 A 2 2 PN 4237003C INDEX m 1023 See channel 1023 A acquisition channels defined 2 13 3 prior to done by system 2 13 4 window creating the 2 13 1 Acquisition card cage communication pathway to
264. arning Labels on Instrument 3 1 5 Unloading the Cytometer 3 2 1 Dual CRT Installation 3 2 3 Rinse Bottle Holder Location 3 2 4 Screws Securing Compressor to Pneumatics Cabinet 3 2 5 Pinch Valve Deactivator Clip 3 2 6 Filter Slots 3 2 6 Sample Collection Compartment 3 2 7 Pinch Valve Deactivator Clip 3 3 1 Electronic Cable Connections 3 4 1 Connections to Workstation CPU 3 4 2 xix XX 3 4 3 3 5 1 3 5 2 3 5 3 3 6 1 3 6 2 3 7 1 Af 3 7 3 3 3 7 5 3 7 6 3 7 7 3 7 8 3 7 9 3 7 10 3 7 11 3 7 12 3 7 13 3 7 14 3 7 15 3 9 1 3 9 2 3 9 3 3 9 4 3 9 5 3 9 6 3 9 7 3 10 1 3 12 1 3 13 1 3 14 1 3 14 2 3 19 1 4 1 1 42 1 4 2 2 4 2 3 4 4 1 4 11 1 4 11 2 4 11 3 4 12 1 4 12 2 4 12 3 4 12 4 4 12 5 4 12 6 412 7 Sheath and Waste Bottles 3 4 4 Fan Locations 3 5 1 Flow Cell Shipping Plug Removal 3 5 2 Deflection Plate 3 5 3 Tube Placement for Sorting 3 6 2 ESP Front and Back Porch Adjustment Knobs 3 6 5 Path of Interlock Cable to Multibus System 3 7 3 Cable Connections for Laser Installation 3 7 4 Argon Laser Installation 3 7 5 HeCd Laser Head Mounting 3 7 7 HeCd Power Supply Connections 3 7 7 HeCd Optical Arrangement 3 7 8 HeCd Laser Configuration 3 7 8 1305 Mounting 3 7 9 Water Cooled Optical Arrangement 3 7 9 Water Cooled Laser Arrangement 3 7 10 Beam Expander Reducer 3 7 11 HeCd Laser Covers 3 7 12 Water Cooled Laser Covers 3 7 13 Warning Label
265. ars on the Cytometer screen b Make sure this process is completed before continuing Press to stop acquisition At the Menu bar select Applications gt gt Utilities The Utilities menu lists a series of options in one column and whether or not these options are PRESENT in a second column Locate the Sort 256 x 256 option and verify the word PRESENT is associated with the Sort 256 x 256 option If this option is listed as PRESENT the 256 Sort upgrade is successfully installed Under Heading 5 1 VERIFICATION INSPECTION PROCEDURE a Complete the instructions under heading Operational Test Cytometer Electronics Optics or Fluidics as written b Complete the instructions under heading Operational Test Sorting Ability as written PN 4237003C CONTENTS IE 4 SERVICE AND REPAIR PROCEDURES 4 1 1 41 OPTICAL ALIGNMENT PROCEDURE 4 1 1 System Alignment 4 1 1 Purpose 4 1 1 Standard Configuration Procedure 4 1 1 Tools Supplies Needed 4 1 1 Procedure 4 1 1 Standard Laser Alignment 4 1 2 Purpose 4 1 2 Tools Supplies Needed 4 1 2 Procedure 4 1 2 Flow Cell Alignment Initial 4 1 3 Beam Translator Equipped System Alignment 4 1 3 Purpose 4 1 3 Tools Supplies Needed 4 1 3 Laser Mounting Procedure 4 1 4 Laser Alignment for Gated Amp Systems Standard Laser Alignment 4 1 4 Collinear Laser Alignment Fine Tuning for Three Lasers 4 1 6 Gated Amp Laser Alignment Time Space Separated Laser Alignment 4 1 7 Camer
266. at procedure as they supplement the precautions listed in this section Electronic WARNING Risk of electric shock Be very careful when operating the instrument with safety interlock bypass jumpers installed as you may be exposed to electric shock Always REMOVE ALL SAFETY INTERLOCK BYPASS JUMPERS after servicing the instrument To protect the operator from personal injury this instrument is equipped with safety interlock switches that turn off the power when the front or rear cover is removed Be very careful if you bypass these safety interlocks and operate the instrument with the covers off WARNING Risk of personal injury Rings or other jewelry can contact exposed electronic components causing personal injury from electronic shock Remove rings and other metal jewelry before performing maintenance or service on the electronic components of the instrument CAUTION Risk of damage to electronic components e If you remove or replace a printed circuit card or electronic component while the power is ON the component may be damaged To prevent damage to delicate electronic components turn OFF the power before removing or replacing printed circuit cards and or components Electrostatic discharge ESD can damage disk drives add in circuit cards and other electronic components If there is a possibility of ESD damage with a procedure then perform that procedure at an ESD workstation or wear an antistatic wrist strap attac
267. ated Amp slot 6 J4 J9 J7 J10 J10 J11 J3 J13 J8 J6 J14 J9 J17 J12 J18 Quad 7 Microsecond Delay 2 slot 8 PMT Gated Amp slot 6 12 J3 J15 J5 J16 J7 J19 Quad 7 Microsecond Delay 1 slot 9 Quad 20 40 60 Delay 1 slot 10 J8 J11 J6 J8 J4 J5 J2 J2 Quad 20 40 60 Delay 1 slot 10 Scat Aux Gated Amp slot 11 J6 J4 J7 J7 J8 J10 J9 J3 J11 J6 J12 J9 J15 J12 J16 Quad 7 Microsecond Delay 1 slot 9 Scat Aux Gated Amp slot 11 10 J3 J13 J5 J14 J7 J17 PMT Gated Amp slot 6 3PMT Sub SW R 1 slot 5 J20 J3 J21 J2 J22 Bundle these cords and continue 3 9 5 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE 3 9 6 Table 3 9 2 Coax Cable Interconnections From To PMT Gated Amp slot 6 3PMT Sub SW R 2 slot 4 J23 24 2 25 J3 3 PMT Sub SW R 1 slot 5 Dual FL SW R slot 3 J5 3 Sub SW R 2 slot 4 Dual FL SW R slot 3 J5 5 PMT Installation J5 J4 4 PMT Installation J5 Scat Aux Gated Amp slot 11 Peak Scatter Mux SW slot 13 J18 J11 Scat Aux Gated Amp slot 11 3PMT Sub SW R3 slot 12 J19 20 2 21 J3 3 PMT Sub SW R3 slot 12 Peak Scatter Mux SW slot 13 J5 J6 J8 J7 J6 J8 J5 3PMT Sub SW R 3 slot 12 Scat CV SW R1 slot 14 J4 Peak Scatter SW slot 13 Scat CV SW R1 slot 14 J2 J6 J3 J5 3 PMT Sub SW R 1 slot 5 Peak Scatter Mux SW slot
268. ated beam shutter and install second alignment target at this location 5 Remove the flow cell tip beam shaping lens sample tubing and sample drawer ATTENTION This alignment must be done very carefully It may be helpful to use a power meter to measure and record the laser power passed through the targets This will give you a reference value to which you should return to when you replace the plate 6 Adjust the mounting hardware of the Argon laser as needed to perfectly center the laser beam through the two target apertures Be sure laser is securely mounted when done 7 Block the laser beam with a tall target or other nonreflecting metallic object Note Be sure not to disturb laser alignment Do not turn laser off 8 Ensure that crystal drive is set to 0 096 and disconnect electrical connector to the Bimorph crystal drive 9 Ensure that deflection is set to 0 096 and a Disconnect the alligator clip from the flow cell sample introduction tube b Disconnect the ground connector from the same cable at the deflection plate holder 10 Using a scribe or permanent marker mark the table top at the edge of the plate to provide a reference for horizontal plate location with reference to the table top 11 Locate the six Allen screw bolts that secure the vertical optical plate to the housing assembly There are four bolts forming a vertical line between the flow cell and lens stages There are two bolts at the lower edge of th
269. ater cooled laser Optional Printer The instruments table can accommodate an optional Printer However the user may prefer to use a separate Printer stand to free up the work space Labels Verify that all labels are in place on the instrument See Figure 3 1 1 Supplies Verify that the necessary supplies are available The supplies include e IsoFlow sheath fluid liters e Fluorospheres e Deionized water liters 4237003C INSTALLATION PROCEDURES 3 PREINSTALLATION CHECK Figure 3 1 1 Warning Labels on Instrument THIS LASER PRODUCT COMFORMS TO THE PROVISIONS OF 21 CFR SUBCHAPTER J SECTION 1040 10 AND 1040 11 VISIBLE AND OR INVISIBLE LASER RADIATION WHEN OPEN AVOID EYE OR SKIN EXPOSURE TO DIRECT OR SCATTERED RADIATION Manufactured DATE 19 by EPICS DIVISION OF COULTER CORP Hialeah FL 33010 7003094A PN 4237003C 3 1 5 INSTALLATION PROCEDURES PREINSTALLATION CHECK 3 1 6 PN 4237003C INSTALLATION PROCEDURES 3 INITIAL SETUP 3 2 INITIAL SETUP Unloading and Moving the Instrument The instrument arrives at the customer site with the Cytometer packed in a palletized box The remainder of the instrument arrives in boxes that are not palletized WARNING Risk of personal injury Do not attempt to unload the main assembly without using a fork lift or a pallet carrier Failure to use a fork lift or pallet carrier can result in serious
270. avelengths with a maximum of about 620 nm Air cooled Argon Laser that provides a wavelength of 488 nm Hoechst Range of wavelengths that overlap the PI emission band UV Laser that provides a wavelength of 351 nm Within Figure 2 6 7 the following are true e The optics are aligned so that the two laser beams intersect the sheath stream The 488 beam upper beam is above the 351 beam The 351 beam lower beam is below the 488 beam Two PMTs and a 550 dichroic filter detect the signals 2 6 6 PN 4237003C 2 INSTRUMENT DESCRIPTION ACQUISITION SUBSYSTEM 2 15280H 82000 end BN gir EM weag 19m07 Z Jase pe ejeg uoN JEUbIS d payey Ie Ad 1 weag 1 49 spied 3429 yeas pue 5 50101 09 x Jase Aq 1 10 Op OZ Z Se wo paAejaq jeubis jeny Aejaq 4 5 vmm cet sx 1 aen a ureag 29567 eufis pacejaq uoN asind 31 V5 sayesauay 101219199 929 UPIM l Du dwy payey CTI 4 MODUM asing IWY
271. ay 1 8 2 8 10 8 2 9 29 6704904 Quad 20 40 60 Microsecond Delay 2 8 2 8 7 6704906 Pulse Generator and Clock 8 2 12 1 6704907 Pulse Generator and Clock 8 2 8 30 8 2 9 9 6704913 Extender Data Acquisition Gated Amp power 8 2 8 15 PN 4237003C 8 PARTS LISTS MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY Table 8 1 4 Cards Continued Part Number Description Figure Item 6704914 Extender Motor Controller power 6704915 Extende r Multibus power 6704916 Extender HV DAC 6704933 Cytometer Computer Backplane 8 2 11 1 6704959 Pulse Pileup Detector TOF 8 2 9 10 6705073 Sample Tube Indicator 6705117 Sensor Interface 8 2 8 16 6705182 Peak Scatter Sensor 8 2 19 9 6705191 Autoclone Sorting Option B 2 25 2 6705246 3 PMT Sub SW R 8 2 8 4 8 2 9 27 34 35 6705249 Dual FL SW R 8 2 8 3 8 2 9 36 6705252 Scat CV SW R 8 2 8 14 8 2 9 25 6705327 Gated Amp Control R3 8 2 9 38 6705328 Interconnect card Autoclone Sorting Option 8 2 23 1 6705329 Position Detection Autoclone Sorting Option 8 2 23 8 6705369 Peak Scatter Mux SW R 8 2 8 13 8 2 9 26 6705400 Peak ADC PSH Control card 8 2 8 24 8 2 9 14 6705504 Pneumatic Interface R2 used with low bleed regulators PN 6232490 and 6232491 6705681 Analyzer Interface 8 2 9 18 6705694 Sort Oscillator R2 8 2 8 18 6705954 Pulse Pileup Detector TOF2 8 2 8 28 8 2 12 5 6706034 Sort Delay 8 2 8 19 6706232 Bitmap and Sort Decision R2 8 2 8 21 8 2 12 12 6706571 Cytometer CPU card with PRO
272. b See Figure 4 12 9 Figure 4 12 9 Laser Beam Centered on Forward Scatter Detector Mask MASK LASER BEAM 7003200A FORWARD SCATTER DETECTOR 4 Atthe Cytometer a Turn the left scope knob clockwise b Select Scope 5 As shown in Figure Figure 4 12 10 select a 1 and FS Int b Trace 2 and PMT2 Peak Figure 4 12 10 Scope Screen Selections SCOPESCREEN Scatter lt gt creen FS PMT2 int Peak Screen axis TRACE DISPLAY Options KZ 7003203A 6 Place a sample of fluorospheres on the instrument and press RUN 7 Center the beam on the absorber in front of the light scatter detector by adjusting the laser beam horizontal knob See Figure Figure 4 12 11 for location PN 4237003C SERVICE AND REPAIR PROCEDURES 4 OPTICAL ALIGNMENT PROCEDURE Figure 4 12 11 Laser Beam Horizontal Adjustment Location FLOW CELL LASER BEAM INTERLOCK VERTICAL VERTICAL DEFEAT BAR ADJUSTMENT ADJUSTMENT FLOW CELL Z AXIS ADJUSTMENT FLOW CELL HORIZONTAL ADJUSTMENT LASER BEAM HORIZONTAL ADJUSTMENT GE LASER SHUTTER 7003196A BEAM SHAPER SCREW 8 Slowly adjust the laser beam horizontal knob in one direction and watch for the horizontal blue line or spot to appear disappear and reappear across the forward scatter detector 9 Adjust the laser beam horizontal knob half way between the two horizontal blue line
273. ble ON Set the Stream control to sort pulse baseline of 0 V 4237003C 4 2 1 4 SERVICE AND REPAIR PROCEDURES SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE 4 2 2 Verify the following with an oscilloscope Front porch length Back porch length 31 25 microseconds Center level length Number of droplets to sort 1 See text for voltage specifications from baseline to center level Front and back porches adjust from 0 to 10096 of center level Tools Supplies Needed Oscilloscope DVM Crystal Drive Test Procedure 1 5 Monitor the signal with a scope at the flow cell where the cable connects to the Bimorph assembly Set Drive Frequency to 32 kHz Set Crystal Drive to 10096 Ensure that the wave form is sinusoidal and measures 160 V peak to peak 5 V with little distortion If the voltage is incorrect do Crystal Drive Adjustment Procedure e If voltage is correct go to step 5 Decrease Crystal Drive setting and ensure the signal reduces proportionately Crystal Drive Adjustment Procedure MU Se ge Verify that the Crystal Drive is at 100 and the Drive Frequency is at 32 kHz Connect the oscilloscope to the Sort Output card 1 Adjust R84 on the Sort Oscillator card for 8 5 V peak to peak Connect the oscilloscope to the bimorph cable at the flow cell Adjust R118 on the Sort Output card for 160 V 5 V peak to peak signal Sort Pulse Amplitude Test Set Drive Fre
274. body 8 2 19 7 8 2 21 1 6704787 Sensor scatter non peak 6705402 Sensor scatter peak R2 8 2 19 9 6705493 Power supply 3000V 8 2 15 2 6705597 Sensor scatter analyzer 8 2 19 9 8 2 20 8 6705987 Deflection plates jet in air flow cell tips PN 4237003C Table 8 1 6 Cytometer Continued MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY PARTS LISTS Part Number Description Figure Item 6854718 Sensor fluid SN4 behind sample station 6856241 Bottle flow cell cleaning 6856505 4 8 2 1 13 8 2 22 1 6856675 Holder deflection body 6856677 Compressor module 115V 6856678 Trap water air water separator 8 2 3 8 6856696 Beamshaper 15 x 60 mm 8 2 19 5 6856703 CRT dual assembly with covers 8 2 1 12 8 2 7 1 6856762 Flow body thin fixed rod old mount 8 2 31 3 8 2 32 3 6856826 Beamshaper 8 x 60 mm 8 2 20 4 6856833 PMT 1 2 3 and 5 8 2 1 14 8 2 22 2 3 4 5 6856842 Drawer sample collection 8 2 1 3 6856866 Door rear transformer rack 8 2 14 1 6856930 Table top 6856967 Vial cap 6856970 Motor mixer 8 2 1 5 6856971 Sample station 8 2 1 4 6856984 Bubble chamber lower 6856985 Bubble chamber upper 8 2 6 1 6857012 Shield CRT housing 6857048 Holder base and end cap sample tube 6857066 Snout pinhole 8 2 20 10 6857133 Shutter assembly 8 2 19 8 2 20 6857170 Blower laser box assembly 6857276 Light source fiber optics PN 3814177 6857369 Bottle rinse 6857388 Valve pinch double action 8 2 19 1 6857400 Holde
275. but separated by 1 16 inch Install the UV dichroic mirror and beam expander Target beam through to target the FLS position Use all four targets Install the 457 dichroic mirror and repeat step 4 with the front laser Do not worry about the beam displacement at this point you will use the for pulses for fine tuning Run beads and establish peak signal pulses Adjust beams to see a 40 microsecond delay Pay special attention to focus You will find that you must compromise the focus to get the best pattern for both beams if the beam heights change greatly during focus 3 9 23 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE 3 9 24 Use targets to block each beam in turn to make sure you know which pulse is associated with which beam Delay appropriate signals establish the window and enable the gated amplifier When running beads the UV power must be high because the beads fluoresce brighter in the 457 nm region When running chromosomes the Hoechst signal is usually much brighter than the Mithromycin so the 457 laser can be run almost at full power and the UV laser must be reduced You can then adjust PMT voltages and so forth for fine tuning Calibration 1 10 11 12 13 Align both lasers with tall targets so both are parallel to each other and the optical table Ensure the beam width at the beam origin and at the wall at least 3 feet away is the same For Elite instruments the front laser
276. by the CPU prior to acquisition to process the correct number of parameters and use the specified timing parameters Once acquisition begins the card cycles through the acquisition process until it receives a command to stop acquisition Controls the acquisition cycle When a PSH card indicates a discriminator satisfied condition then the cycle starts PN 4237003C 2 13 7 2 INSTRUMENT DESCRIPTION SIGNAL SELECTION AND DIGITIZATION Note In a Gated Amp system this can also originate from the Gated Window pulse The and PSH Control card allows all active PSH cards to charge to the maximum input level then closes the PSH inputs to prevent corruption of the stored values Refer to ADC and PSH Timing for details The ADC and PSH Control card receives analog data stretch pulses from the PSH cards and digitizes these inputs one at a time and sequentially places the digital values on the data path bus When the cycle is completed depending on the number of pars being converted the PSH storage capacitors discharge and the PSH inputs reopen to prepare for the next event ADC and PSH Timing The Elite uses two different ADC and PSH Control cards as listed in the following chart Date of ASD Operation System System Manufacture Card Used Supported by Card Elite Before 09 93 ADC and PSH Control No Elite ESP 09 93 and after or any Peak ADC Yes field upgraded system The cards differ in
277. cat Aux Gated Amp slot 11 J2 PMT1 ac Filter Coax Assembly PN 6586479 7 ac Filter Coax Assembly PN 6586479 7 Scat Aux Gated Amp slot 11 J3 PMT2 PMT Gated Amp slot 6 J4 PMT3 PMT Gated Amp slot 6 J5 PMT4 PMT Gated Amp slot 6 J6 5 Gated slot 6 J7 If applicable Non Gated Amp Units Interconnections from Sensor Area FALS WM400 Peak Scatter Mux SW R slot 10 J11 PMT1 ac Filter Coax Assembly PN 6586479 7 ac Filter Coax Assembly PN 6586479 7 Scat CV slot 9 J1 PMT2 3 PMT Sub slot 7 J1 4 10 9 4 SERVICE AND REPAIR PROCEDURES ELECTRONIC ADJUSTMENTS CABLE AND JUMPER CONFIGURATIONS 4 10 10 Table 4 10 6 Cable Connections Continued From To 3 PMT Sub slot 7 J2 4 3 PMT Sub slot 7 J3 Table 4 10 7 Cable Connectors From To P99 Sort Transistor Bkt J2 P59 Dual Crt Controller J2 P61 Dual Crt Controller J3 J43 Dual Crt Controller J9 P60 Digiscope J2 P62 Digiscope J3 J44 Camera Interface J5 P95 Camera Interface J6 Camera Interface Cable Camera Interface J1 P49 Dual Laser Controller J3 P89 Dual Laser Controller J4 4237003C SERVICE AND REPAIR PROCEDURES SORT SENSE FLOW CELL ADJUSTMENT AND VERIFICATION 4 11 SORT SENSE FLOW CELL ADJUSTMENT AND VERIFICATION Purpose Use this procedure to adjust the sort sense flo
278. ccessory lasers may require factory certification of the installer Do not install any laser unless you have the required training and or certification The alignment procedures depend upon the type of lasers installed and their optical configuration Refer to the correct laser alignment procedure for details Argon Laser 1 Turn ON the Argon laser if present 2 Verify operation of the laser cooling fan 3 Verify light output from the laser which is approximately 20 second delays HeNe Laser 1 Turnon the HeNe laser if present 2 Verify operation of the laser cooling fan 3 Verify light output from the laser Optional Lasers Inspect all optional lasers for correct operation Refer to the user or service manual supplied with the laser for specifications and procedures Computer 1 Turn ON the upper power switch on the front of the Cytometer 2 Ensure that the Elite menu appears after bootup 3 Test the system for known software viruses by using the latest available version of virus detection software Alignment Check Do the applicable procedures under Heading 4 1 OPTICAL ALIGNMENT PROCEDURE 3 5 4 PN 4237003 3 6 PN 42370036 INSTALLATION PROCEDURES SYSTEM TESTING SYSTEM TESTING Acquisition Check 1 Create a protocol to acquire fluorospheres as follows e Histogram 1 FALS vs 9015 stop at 5 000 Histogram 2 FALS Histogram 3 9015 e Histogram 4 Log GFL PMT2 e Histogram 5 GFL e Histogram 6
279. cess takes several minutes 12 Under Heading 5 1 VERIFICATION INSPECTION PROCEDURE complete the instructions under heading Operational Test Cytometer Electronics Optics or Fluidics as written 4 13 2 PN 4237003 5 MAINTENANCE PROCEDURES 5 1 1 5 1 VERIFICATION INSPECTION PROCEDURE 5 1 1 Verifying Instrument Performance Every Call 5 1 1 Cytometer 5 1 1 Workstation 5 1 1 System Not Serviced Within Past 30 Days 5 1 1 Operational Test Cytometer Electronics Optics or Fluidics 5 1 1 Operational Test Sorting Ability 5 1 2 Operational Test Every Call 5 1 2 TOF Verification 5 1 2 Purpose 5 1 2 Procedure 5 1 2 5 2 PREVENTIVE MAINTENANCE PMI PROCEDURE 5 2 1 Purpose 5 2 1 Tools Supplies Needed 5 2 1 Before You Begin 5 2 1 Procedure 5 2 1 PN 4237003C 5 1 5 1 4237003C MAINTENANCE PROCEDURES 9 1 VERIFICATION INSPECTION PROCEDURE Verifying Instrument Performance Every Call Cytometer 1 Ensure all fixed panels and covers are in place 2 Ensure all moveable covers operate properly 3 Inspect all fans Two on Multibus card cage One above Gated Amp card cage e Two on right rear panel e One on left rear panel 4 Inspect sampling and collection areas for leakage and spillage Clean and repair as needed Inspect flow cell area for saline spray deposits and corrosion Clean and repair as needed Inspect air water separator Workstation 1 2 Go t
280. chrome Helium Cadmium 74 Laser 1 2 5 1 2 5 Laser Warning Labels Coherent Innova Enterprise Laser 1 2 5 1 2 6 Laser Warning Labels Coherent Innova 305 Laser 1 2 6 1 2 7 Laser Warning Labels Coherent Spectrum Laser 1 2 6 1 2 8 Laser Warning Labels Uniphase Red HeNe Laser 1 2 7 1 2 9 Laser Warning Labels Melles Griot Green HeNe Laser 1 2 7 1 2 10 Laser Warning Labels Sensing Area 1 2 8 PN 4237003 1 1 1 4237003C INTRODUCTION 1 1 MANUAL DESCRIPTION Scope This manual provides the reference information and procedures needed to service and maintain the COULTER EPICS Elite flow cytometer and the COULTER EPICS Elite ESP flow cytometer It is available both online and in hard copy The online manual is released on the Service Resource Kit CD ROM PN 6417471 1 This manual does not contain information and or procedures already covered in other manuals for these instruments Use this manual in conjunction with the following Elite customer manuals Reference manual PN 4235904 Operators Guide 4235932 e Options manual PN 4235965 Software manual PN 4237024 e Special Procedures and Troubleshooting manual PN 4235931 To service the Computer Workstation which consists of the central processing unit CPU color monitor keyboard and mouse refer to the manual supplied by the manufacturer of the Workstation To service the Graphic Printer refer to the manual supplied by the manufactur
281. cket mount pinch valve analyzer 8 2 18 22 6912841 Hardware laser mounting Innova 90 5 6912844 Hardware laser mounting Innova 305 6912869 Autoclone Sorting Option Sorting Option 6912870 Hardware laser mounting Cyonics Argon Laser 6912871 Hardware laser mounting HeNe red Uniphase 6913205 Hardware laser mounting Enterprise Innova 69132278 Hardware laser mounting Spectrum Table 8 1 11 Kits Part Number Description Figure Item 6912185 He Cad Expander 6912268 Filters Biohazard replacement 6912655 HeCd Indo optics 6912749 HeNe red Uniphase 8 2 36 1 6912750 Argon laser Cyonics 8 2 36 2 6912792 Innova 305 8 2 36 3 6912795 Piezo flow cell mount 6912797 PMT 5 upgrade before SN U07122 6912798 Gated Amp upgrade after SN 007122 6912801 HeCd laser 6912827 Gated Amp upgrade before SN U07122 6912831 Innova 305 water cooled laser 6912832 90 5 water cooled laser 6912833 Cover upgrade 6912834 Optics HeCd 74 laser 8 2 38 1 6912938 Filters sheath and waste 0 2 micron 6912971 He Cad Omnichrome mounting hardware 6912985 Beam translator with laser 8 2 35 9 6913056 PMT 5 switch amps 6913251 Snout upgrade 6913419 PMI Service kit 6913199 Enterprise laser Coherent PN 4237003C 8 1 15 8 PARTS LISTS MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY Table 8 1 11 Kits Continued Part Number Description Figure Item 6913206 Enterprise optics 6913211 Spectrum laser Co
282. crosecond Delay 1 card jumpers 4 10 3 A 2 3 Quad 20 40 60 Microsecond Delay 2 card jumpers 4 10 3 A 2 3 Quad 20 40 60 Microsecond Delay card jumpers A 2 3 Quad 7 Microsecond Delay 1 card jumpers 4 10 3 Quad 7 Microsecond Delay 2 card jumpers 4 10 3 A 2 3 4 10 3 A 2 3 Quad 7 Microsecond Delay card jumpers 3 9 4 Quad PSH 1 card jumpers 4 10 3 A 2 3 Quad PSH 2 card jumpers 4 10 3 A 2 3 Quad PSH card block diagram 2 13 3 2 13 5 R ramps use during unloading 3 2 2 ratio calculation of 2 13 5 read pressure See pneumatics regulators pneumatic See pneumatics requirements Cytometer installation 3 2 1 sample 2 1 2 sorting 3 6 1 requirements initial installation ambient temperature 3 1 3 customer training 3 1 1 drainage 3 1 3 heat dissipation into room 3 1 3 humidity 3 1 3 labels placement of 3 1 4 operating clearances 3 1 4 power 3 1 1 power outlets 3 1 2 Printer optional 3 1 4 space 3 1 3 supplies needed for installation 3 1 4 temperature stability 3 1 3 water for Coherent 305 Laser 3 1 2 rinse flow valve VL11 function of 2 9 2 INDEX 13 INDEX rinse pressure valve VL8 function of 2 9 2 S safety concerns description of 1 2 1 safety precautions biological 1 2 1 electronic 1 2 1 laser 1 2 2 what to tell the customer 1 2 9 sample collect drain valve VL10 function of 2 9 2 sample collection compartment installation procedure 3 2 7 sample pinch valve
283. ct mapped write back cache Bus ISA Local bus PCI 32 bit at 33 MHz Hard drive 1 2 gigabyte or higher enhanced IDE with Mode 3 timing Controller Tekram PCI caching controller with 4 MB cache transfer rates up to 8 megabytes per second Floppy drives One 3 5 in 1 44 megabyte and one 5 25 in 1 2 megabyte Video SVGA with 2 megabytes standard PCI local bus 1024x1280 resolution 16 million colors Expansion slots 4 ISA slots 2 PCI slots and 1 PCI ISA selectable slot Adapter card Dual serial and one parallel ISA Adapter card provides parallel port for printer BIOS Flash BIOS American Megatrends provides PCI plug and play PN 4237003C 3 21 1 PC MODEL PENTIUM 133 MHz UPGRADE PC MODEL PENTIUM 133 MHz UPGRADE Table 3 21 1 Technical Specifications of the DTX Pentium Computer Continued Item Specifications Power supply 200 W 115 230 V selectable 50 60 Hz Regulatory Safety compliances Us UL 1950 tst edition e Canada CSA C 22 2 No 950M 89 Europe TUV to EN 60950 with ZHI 618 EMI RF US FCC CFR 47 Part 15 Level B e Canada DOC CRC c 1374 Class B Europe VDE 0871 Class B CISPR B Tools Supplies Needed Elite Pentium Full Workstation Upgrade Kit PN 6915005 which includes gt Elite DTX Pentium computer with SVGA video board and 16 RAM gt Serial mouse gt Keyboard gt 17 in Sony color monitor 1024x1280 resolution gt Elite software lib
284. ction 2 9 2 VL9 function 2 9 2 VL10 function 2 9 2 VL11 function 2 9 2 VL12 function 2 9 2 VL13 function 2 9 2 VL14 function 2 9 2 VL15 function 2 9 2 voltages Bertans power supply produced by 2 5 8 DAC reference measurement of 2 9 4 error definition of 2 9 5 outputs held by peak sense and hold circuit 2 13 3 ratio routing of 2 13 5 INDEX 17 INDEX W waste bottle full error message when 3 19 2 modification procedure 3 19 3 overflow result of 2 9 1 Waste Filter Error 2 9 1 watchdog C44 Sensor Interface card location of 3 17 1 circuit description of 2 5 8 R77 Sensor Interface card location of 3 17 1 signal relay function of 2 9 4 timing when to do 3 17 1 timing See timing waveforms sort adjusting and verifying 4 2 1 adjusting the crystal drive 4 2 2 adjusting the sort pulse amplitude 4 2 3 delay settings 4 2 3 phase settings 4 2 3 setting the crystal drive 4 2 2 testing the sort pulse amplitude 4 2 2 A 3 1 verifying synchronicity of sort pulses with crystal drive 4 2 4 verifying the sort function 4 2 4 window pulse See gate window window sort description 2 8 11 Z zero value See 0 zero value defined zooming the camera See camera 18 INDEX PN 4237003C TRADEMARKS m The BECKMAN COULTER logo COULTER CYTO TROL EPICS IsoFlow and MDADS are trademarks of Beckman Coulter Inc All other trademarks service marks products or services are trademar
285. d Left Pulses Mux and Scope card Scatterplot Mux and Scope card Graphs Cytometer CPU card Camera Camera Figure 2 5 2 shows the pathways for the various display sources as they are processed and selected for display Figure 2 5 2 Block Diagram Video Subsystem CAMERA INTERFACE CARD IMAGE MEMORIES CONTROLS amp SYNC SYNC FOR CAMERA J9 VIDEO DISPLAY GENERATION LEFT SCREEN GRAPHS HISTOGRAMS SERVICE RIGHT SCREEN MENU SCREENS DUAL CRT CARD 2 5 4 DISPLAY SELECT KNOB UNDER LEFT CRT SCOPE CIRCUITS MEMORY DOT CLOCK CONTROL LOGIC SWEEP GENERATOR GRID GENERATOR SCATTERPLOT CAMERA MUX amp SCOPE CARD CONTROL SCREENS MENUS SYNC KNOB 7003025A PN 4237003C PN 42370036 INSTRUMENT DESCRIPTION CONTROL AND DISPLAY SUBSYSTEMS Camera Image Signal Flow Video Display The camera video output goes directly to the Camera Interface card via a DIN cable The synchronization signals horizontal and vertical sync for the camera are generated on the Dual CRT Control card and are supplied to the camera through the Camera Interface card Camera Interface Card The video image that the camera receives is digitized and temporarily stored in high speed memory on the Camera Interface card This allows the Cytometer s CPU to manipulate the camera image The pan feature is implemented by selecting the portion
286. d 2 8 10 Sort Envelope 2 8 11 Block Diagram Sort Output Card and Transistor Bracket Card 2 8 14 Interconnection of Pulse Pileup Det TOF Card 2 8 15 TOF Function of Pulse Pileup Det TOF Card 2 8 16 PPU Function of Pulse Pileup Det TOF Card 2 8 18 Power Supplies Location 2 8 20 Valve Operation 2 9 3 Block Diagram Sample Pressure System Old Type 2 9 6 Block Diagram Sample Pressure System New Type 2 9 7 Block Diagram Sheath Pressure System Old Type 2 9 9 Block Diagram Sheath Pressure System New Type 2 9 11 Block Diagram Compressor Module 2 9 12 3 PMT Sub SW R Card Block Diagram 2 10 1 Dual FL SW R Card Block Diagram 2 10 2 Scat CV SW R Card Block Diagram 2 10 3 Block Diagram Peak Scatter Mux SW R Card 2 10 4 Block Diagram Scat CV Amp Card 2 11 1 Block Diagram 3 PMT Sub Amp Card 2 11 2 Block Diagram Dual FL Amp Card 2 11 3 Block Diagram Gated Amp Control Card 2 12 1 Block Diagram Scat Aux Gated Amp Card 2 12 3 Block Diagram PMT Gated Amp Card 2 12 5 Block Diagram 20 40 60 Microsecond Delay Card 2 12 6 Block Diagram 7 Microsecond Delay Card 2 12 6 Block Diagram Mux and Scope Card 2 13 3 Block Diagram PSH Card s 2 13 5 Block Diagram ADC and PSH Control Card 2 13 6 Block Diagram Peak ADC PSH Control Card 2 13 7 ASD Settings Diagram 2 13 9 Peak ADC PSH Card Timing 2 13 10 ASD Settings Diagram 2 13 11 Old ADC Timing Diagram 2 13 12 Block Diagram Lister Cards 2 13 13 W
287. d focus on instrument setup filters and sample preparation This test can be performed for the other PMTs by moving the filters as needed Note that the test is always done with the PEAK PMT signal Although subtraction is also seen on the Integral and log histograms the reduction does not relate directly to the subtraction percent especially for log Do the electronic test by providing the same pulse to two and then observe the histograms as subtraction is used In this example PMT2 and PMT3 subtraction is tested You can use the same procedure with appropriate adjustments to test all subtraction combinations PMT 3 PMT 2 Compensation 1 At the Cytometer power down Sheath Vacuum and Valves 2 Turnlasers OFE 3 Configure the filters as shown in Figure 7 2 2 Figure 7 2 2 Filter Configuration BLANK NO FILTER C 7003081A OR MIRROR At the Cytometer Options screen set Align LED to Pulsed 0 25kHz and 100 5 Create a protocol to acquire PMT2 peak and PMT3 peak histograms as follows Parameters PMT2 Peak PMT3 Peak Histograms PMT2 Peak PMT3 Peak PMT2 x PMT3 Peak PMT2 discriminator 100 PMT2 gain PMT3 gain 20 color compensation to 096 initial PN 4237003C 7 2 3 TROUBLESHOOTING TROUBLESHOOTING THE ACQUISITION SUBSYSTEM 7 2 4 Begin acquisition and adjust PMT2 and so the mean channel of both histograms is at channel 512 Test subtraction a Set PMT3 PMT2 subtract P
288. d Amp card cage Two on the Multibus card cage One at the rear of the Cytometer One at the back of the sort collection drawer the fan is activated by the adjacent switch Figure 3 5 1 Fan Locations PN 4237003C 7003090A 3 5 1 INSTALLATION PROCEDURES TESTING ASSEMBLIES Compressor Module l Open the sheath and waste compartment door 2 Disconnect the center blue quick connect fitting from the lower front of the Cytometer 3 Verify the system pressure and the compressor pressure is correct a Open the pneumatics compartment door b Ensure the system pressure reads 30 psi Adjust the regulator as necessary c Ensure the Compressor modules pressure reads 50 psi Adjust the regulator as necessary 4 Reconnect the center blue quick connect fitting to the lower front of the Cytometer Pneumatics 1 Remove the shipping plug from the flow cell as shown in Figure 3 5 2 Figure 3 5 2 Flow Cell Shipping Plug Removal FLOW CELL SHIPPING PLUG 7003137A 2 Install the Biosense tip 3 5 2 PN 4237003C 4237003 INSTALLATION PROCEDURES TESTING ASSEMBLIES 3 Atthe Cytometer touch screen select a Power Up Valves b DEBUBBLE three times c Shutdown Replace the BioSense tip with the sort sense tip At the Cytometer touch screen select Power Up Valves Press CLEAR and DEBUBBLE as many times as necessary to obtain a stable sheath stream Examine the flow cell closely f
289. d stream in the wells with zero offsets Tray does not rotate exactly 180 degrees during operation Setscrew that secures the tray holder to the stepper motor shaft is loose Tighten the setscrew through the upper hole in the front of the arm Rotation speed is incorrect Adjust speed as needed to obtain smooth rotation without overshoot Incorrect movement LED markers do not appear as expected 1 Check optical sensor and encoding strip for dust or contamination Wipe clean as needed 2 Verify operation of sensor by manually moving the arm assembly while reading the marker box on the Autoclone Sorting Option Control screen 3 Observe the LED markers on the Autoclone Sorting Option card e f the markers indicate the Autoclone Sorting Option arm position correctly then the problem is with communication between the Autoclone Sorting Option card and the Cytometer computer Check the ribbon cable that connects the Autoclone Sorting Option card to the host system e f the markers do not indicate the Autoclone Sorting Option arm position correctly then check the following gt cable gt Position Detect card Interconnect card gt Main cable Autoclone Sorting Option card No movement No power to motors Check all power supplies Adjust or replace as needed Waste catcher beak overflows Insufficient vacuum from compressor Check compressor output Repair or replace
290. ddress and data are common to all card slots there is a set of priority lines that are not This means that the CPU card must always reside in slot 1 top but nonprocessing cards will work in any available slot The Multibus subsystem consists of the e Multibus CPU card e External Memory or 256K Memory card Data Taker Interface card e Serial I O card e Camera Interface card Digiscope card Dual CRT Control card Dual Laser Control card These cards are grouped by subsystem and are discussed in Heading 2 9 Cytometer Computer Subsystem This subsystem consists of the Multibus CPU card e External Memory or 256K Memory card e Serial I O card Multibus CPU The Multibus CPU card Executes the program stored in the EPROM and the battery backed RAM that are on the CPU and the External Memory or 256K Memory e Provides a user interface with the CRT as output e Accepts input from the touch screen and membrane pneumatic panel Communicates with the Workstation through one of the RS232 ports on the Serial Interface card PN 4237003 2 5 1 2 INSTRUMENT DESCRIPTION CONTROL AND DISPLAY SUBSYSTEMS Controls the air cooled lasers directly through the Multibus and the Dual Laser Control card Controls the two CRT displays through the Digiscope Dual CRT Control and Camera Interface cards See Video Subsystem Cytometer for more information on these cards Connects to the Pneumatic Interface card
291. de but it is not limited to protective eyewear gloves and suitable laboratory attire when operating or maintaining this or any other automated laboratory analyzer This Service Manual contains confidential information of Beckman Coulter Inc and its receipt or possession does not convey any rights to reproduce disclose its contents or to manufacture use or sell anything it may describe Reproduction disclosure or use without specific written authorization of Beckman Coulter Inc is strictly forbidden Copyright Beckman Coulter Inc 1991 2001 All rights reserved REVISION STATUS a Initial Issue System Installation Procedure 11 91 Software Version 2 0 Revision B Complete Service Manual 04 97 Released by CN 303160 0994 Software Version 4 01 Revision C 12 01 Released by CN 303160 1702 Software Version 4 5 This manual was reformatted to enhance the electronic version Reformatting the manual changed the appearance of each page but it did not change the content Consequently a revision C manual and a revision B manual with revision C change pages look different but they contain the same material page for page The material in the revision C change pages was updated for hardware and software changes made since the last revision including the Elite 256 Sort option Revision C is being released as change pages PN 4277432 This revision updates and incorporates information released by Service Memos 1382 and 1795 and
292. der PMT4 3 9 13 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE 06100 Duupadde asouy 19349 950049 sur 5105095 y yead pue Camp xne 29 9 99e1 Aejap pue upi 909 199 95 918 pares inejap jur zxne adoos sd j919jduuo2 29 xne sa21nos eubis osip n s dwe paye uaap TIVOAY 10 JAYS U9n0 7 19 99u99s9J0Jn j e pue jou Jo 099195 SUE 0 uono Lens smoys sasind yeed 19 0 Weag Jaddn 9 euim 12984 Pinoys 391049 Aejap 2 09 40 295 Qr en QZ I SN 5901000 9L 01 dn 2994 pue anes 0 NOA sung uonounj e gau SONILLAS L10v43d TIv93u TI V93U JAVYS U9919S NIYIN sKe dsip pajoe as uauM HEET SNOILdO sKe dsip payoajas uau M shejdsip pejoe as UIAM 9919 9155 03159 093 payogjas au 9919 sKe dsip pe ejdsip uauM AV T3Q 1VN9SIS Jaye o moys And Buros SIU pajqeua uauM ON ureaq Jaddn ay e Ajuo yng 921 05 11294 4104 Aq
293. diation with a finite size its wavelength According to the wave propagation theory the spot size that a laser beam can be focused to is _ D K where constant related to lens properties wavelength of laser f focal length of lens D diameter of laser beam Beam shapers use a combination of two cylindrical lenses that are made with one flat surface and one cylindrical surface there is no spherical surface on a beam shaper Since the glass curves in only one direction refraction occurs in only one direction By using two oppositely oriented lenses it is possible to tailor the horizontal and vertical spot sizes Figure 2 7 6 shows the beam shaper s optics 2 1 5 INSTRUMENT DESCRIPTION OPTICAL ELEMENTS Figure 2 7 6 Beam Shaping Optics Table 2 7 1 shows some typical applications for different beam shapers SIDE VIEW TOP VIEW VERTICAL LENS FOCAL LENGTH HORIZONTAL LENS FOCAL LENGTH P 7003176A Table 2 7 1 Typical Applications of Beam Shapers Beam Shaper DISCSAT Band PPU Data Sorting Application Flow Cell Laser Power Spot Size EXT Width Sensitivity Rate Lymphocytes with 76 um 200 mW 15 x 33 um 2 5 msec High 0 5 4 0 High high intensity staining jet in air or water cooled umsec fast quartz Lymphocytes with low 76 um sort 15 mW 15 x 60 um 10 umsec Low 10 20 Low inte
294. ds an error signal to the Pneumatics Control card and the Motor Control card These cards respond to this error by blocking the 24 V from the Camera Motor Control circuit the solenoid drive circuits and the pressure regulator drive circuits This action prevents damage to the system in the event of Multibus CPU card failure Motor Control Card The Motor Control card Controls the camera s movement and zoom motors The Multibus CPU card programs special dc motor control integrated circuits on this card for the desired position or zoom These circuits then activate the motors until the correct feedback voltage is obtained from the potentiometers Provides power to the mixer motor for fast and slow speeds The speed control potentiometer on the front of the unit is wired in series with the mixer motor Reads the tube sensor mechanism in the sample station DAC Card The DAC card receives the desired PMT voltages from the Data Taker Interface card Next the DAC card converts the digital values to analog voltages and sends them to the proper PMT power supply Bertans The Bertans power supply produces a voltage output of up to 2 000 V 2 5 8 42370036 INSTRUMENT DESCRIPTION ACQUISITION SUBSYSTEM 2 6 ACQUISITION SUBSYSTEM Analog Signal Processing The sensors PMTs and scatter detector generate signals that are processed to develop the signals that the Mux and Scope card selects for display acquisition and or sorting
295. e Keep the laser shutter closed and or block laser beams except when needed Perform all alignments at minimum power Align the UV by wearing protective eyewear and using a business card to observe fluorescence from the laser Avoid skin exposure to any laser beam Always consider where the laser beams may go before opening the shutter PN 4237003C 3 14 1 INSTALLATION PROCEDURES ENTERPRISE LASER UPGRADE 3 14 2 Important Operating Notes Optimum laser lifetime is achieved with the laser operated at 50 mW UV in UV regulation When the heat exchanger is used be sure the hot air exhaust from the exchanger is isolated from the laser head and the Elite system Ensure lab HVAC system can maintain a stable ambient temperature within specified range Refer to the Coherent Enterprise laser manual s For best power regulation lowest noise and best laser life the manufacturer recommends operation at 50 mW UV power continually The ND attenuator can be used to reduce the 488 power levels if desired If the laser is operated at reduced power levels it is important that it be operated at the high power level for one hour at least once a week You should discuss the different laser operating modes with the customer to determine how to best meet their needs The laser can be operated in either UV or 488 light regulation For single beam applications use the appropriate mode Note that 488 mode is a secondary control loop the 488 sensor ou
296. e card Pneumatics Control See Pneumatics Control card Position Detection See Position Detection card Prism and Sort Window Test See Prism and Sort Window Test card Pulse Generator See Pulse Generator card Pulse Generator and Clock R See Pulse Generator and Clock R card Pulse Pileup Det TOF See Pulse Pileup Det TOF Pulse Pileup Det TOF See Pulse Pileup Det TOF card Quad 7 Microsecond Delay See Quad 7 Microsecond Delay card Quad PSH 1 See Quad PSH 1 card Quad PSH 2 See Quad PSH 2 card Quad PSH See Quad PSH card requirements for sorting 3 13 2 Scat Aux Gated Amp See Scat Aux Gated Amp card Scat Aux Gated Amp R See Scat Aux Gated Amp R card Scat CV Amp SW R See Scat CV Amp SW R card Scat CV Amp See Scat CV Amp card Scat CV SW R See Scat CV SW R card Scat CV SW R1 See Scat CV SW R1 card Scatter Sensor R Pre Amp See Scatter Sensor R Pre Amp card Sensor I O See Sensor I O card Sensor Interface See Sensor Interface card Serial I O See Serial card Sort Delay See Sort Delay card Sort Delay R3 See Sort Delay R3 card Sort Oscillator See Sort Oscillator card Sort Oscillator R2 See Sort Oscillator R2 card Sort Output See Sort Output card summary of upgrades for ESP instruments 3 13 5 Transistor Bracket See Transistor Bracket card circuit discriminator range 0 to 10 V 2 13 1 circuit analog calculator PN 42370036 INDEX defined 2 13 5 circuit discriminator function of
297. e 3 drop sort envelope on the oscilloscope Slowly increase the delay between the pulses with R109 while observing the sort pulse You will see the center level of the 3 drop envelope stretch as the pulses move further apart this verifies the sort pulse extension function Sort Pulse Truncation Test This test verifies that an unwanted cell following a good cell can be aborted by shortening the sort pulse envelope For this test the two different integral PMT2 signals will simulate the good and bad cell 1 Adjust R53 on the Pulse Generator card so the second 2 Int pulses is about one division smaller than the first pulse You are actually adjusting the second pulses width so the PMT 2 Peak pulses will remain at about the same height Adjust R109 for a 120 microsecond delay between pulses Acquire the pulses and ensure that you get two populations on the PMT2 int histogram Create two new sort regions as wide as possible on the PMT2 integral histogram a Put region B around the left dimmer population b Put region C around the right brighter population At the Sort screen change it to sort region B to the left and region C to the right 4237003C SERVICE AND REPAIR PROCEDURES 4 SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE Send the new sort regions to the Cytometer Turn Sort Right Enable ON and Sort Left Enable OFF Set Coinc Abort to ON and Drops Sorted to 3 Start Sort and observe sort pulse on oscil
298. e 8 2 16 8 2 16 Analyzer Sample Drawer See Table 8 2 17 8 2 17 Analyzer Top Inside View See Table 8 2 18 8 2 18 Flow Cell Area Elite See Table 8 2 19 8 2 19 Flow Cell Area Elite Analyzer See Table 8 2 20 8 2 20 Deflection Body and Holder See Table 8 2 21 8 2 21 Photomultiplier Tubes PMTs See Table 8 2 22 8 2 22 Autoclone Sorting Option Mechanism See Table 8 2 23 8 2 23 Autoclone Sorting Option Beak Waste Catcher See Table 8 2 24 8 2 24 Autoclone Sorting Option Location and Connections See Table 8 2 25 8 2 25 Autoclone Sorting Option Accessories See Table 8 2 26 8 2 26 Transformer Drawer Assembly See Table 8 2 27 8 2 27 Computer Box Assembly See Table 8 2 28 8 2 28 Beam Splitter Dichroic See Table 8 2 29 8 2 29 Beam Shaper See Table 8 2 30 8 2 30 Flow Cell Stage Elite ESP See Table 8 2 31 8 2 31 Flow Cell Stage Elite See Table 8 2 32 8 2 32 Model 74 HeCd Laser Mounting Hardware See Table 8 2 33 8 2 33 Argon Laser Mounting Hardware See Table 8 2 34 8 2 34 HeNe Laser Mounting Hardware See Table 8 2 35 8 2 35 Laser Placement Top View See Table 8 2 36 8 2 36 Cytometer Inside Right Rear View See Table 8 2 37 8 2 37 Omnichrome HeCd Laser See Table 8 2 38 8 2 38 Autoclone Sorting Option 8 1 1 Beam Shapers 8 1 2 8 1 8 1 3 8 1 4 8 1 5 8 1 6 8 1 7 8 1 8 8 1 9 8 1 10 8 1 11 8 1 12 8 1 13 8 1 14 8 1 15 8 1 16 8 1 17 8 1 18 8 1
299. e Holder Location 3 2 4 Screws Securing Compressor to Pneumatics Cabinet 3 2 5 Pinch Valve Deactivator Clip 3 2 6 Filter Slots 3 2 6 Sample Collection Compartment 3 2 7 Pinch Valve Deactivator Clip 3 3 1 Electronic Cable Connections 3 4 1 Connections to Workstation CPU 3 4 2 Sheath and Waste Bottles 3 4 4 Fan Locations 3 5 1 Flow Cell Shipping Plug Removal 3 5 2 Deflection Plate 3 5 3 Tube Placement for Sorting 3 6 2 ESP Front and Back Porch Adjustment Knobs 3 6 5 Path of Interlock Cable to Multibus System 3 7 3 Cable Connections for Laser Installation 3 7 4 Argon Laser Installation 3 7 5 HeCd Laser Head Mounting 3 7 7 HeCd Power Supply Connections 3 7 7 HeCd Optical Arrangement 3 7 8 HeCd Laser Configuration 3 7 8 1305 Mounting 3 7 9 Water Cooled Optical Arrangement 3 7 9 Water Cooled Laser Arrangement 3 7 10 42370036 3 3 7 11 Beam Expander Reducer 3 7 11 3 7 12 HeCd Laser Covers 3 7 12 3 7 13 Water Cooled Laser Covers 3 7 13 3 7 14 Warning Labels in Sensing Area 3 7 14 3 7 15 Warning Labels Rear of Instrument 3 7 15 3 9 1 Anti Static Container of Gated Amp Kit 3 9 1 3 9 2 Jumpers and Scope Card 3 9 3 3 9 3 Gated Amplifier Assignment Screen 3 9 9 3 9 4 Gated Amplifier Assignment and Adjustment Potentiometer Locations 3 9 11 3 9 5 Gated Amplifier Control Screen 3 9 14 3 9 6 Gated Amp Assignment Screen 3 9 15 3 9 7 Standard Laser Configuration 3
300. e If Autoclone Sorting Option arm is level but there is no clearance between the bottom of the table and the top of the arm then adjust optical table for about 1 8 in clearance Loosely fasten the Autoclone Sorting Option mechanism with screws and washers to the frame Note Be sure to leave access to fan assembly cable Connect 37 pin D connector of cable assembly to Autoclone Sorting Option Interconnect card Calibration Procedure 1 2 10 Turn power ON and install new software See Heading 4 9 SOFTWARE INSTALLATION At the Cytometer screen select Options gt gt Autoclone Sorting Option Control gt gt Adjust Screen gt Calibrate gt gt Load Verify that after certain movements the Autoclone Sorting Option stops at the load position Place the tray holder into arm with notch toward the front and the tray parallel to optical table and secure with set screw Place 96 well tray into holder with letters to front Select Load at the Cytometer screen Verify that the arm moves and that when it stops the sheath is in center of well A12 If not adjust screws securing the Autoclone Sorting Option mechanism Note Press SHEATH to determine location of stream then VACUUM to stop flow Do not overflow the sample tray Select the A1 position on the screen to move tray to selected location Adjust the unit so that the sheath is in center of well A1 Repeat steps 6 and 7 as necessary to align tray Select position H1
301. e J9 P76 Pin 1 on top Gated Amp Control J1 P69 Pin 1 on top CPU J1 P80 Pin 1 to right PN 4237003C PN 42370036 ELECTRONIC ADJUSTMENTS CABLE AND JUMPER CONFIGURATIONS Table 4 10 6 Cable Connections Continued SERVICE AND REPAIR PROCEDURES From To CPU Card J2 P92 Pin 1 to left Serial 1 0 J1 P74 Pin 1 to left Serial 1 0 J2 P93 Pin 1 to left Digiscope P87 Pin 1 to right Data Taker Interface J1 P37 Pin 1 to right Data Taker Interface J2 P84 Pin 1 to right Sort Bracket J1 P2 Sort Osc R2 Card Other Cables Dual CRT Power Cable P65 Position P65 on Dual CRT Pivot Mount Mux and Scope Card Cable Connections Mux and Scope J1 Pulse Pileup Det TOF J1 Mux and Scope J2 Quad PSH 1 J3 Mux and Scope J3 Quad PSH 1 J5 Mux and Scope J4 Quad PSH 1 J7 Mux and Scope J5 Quad PSH 2 J1 Mux and Scope J6 Quad PSH 2 J3 Mux and Scope J7 Quad PSH 2 J5 Mux and Scope J8 Pulse Pileup Det TOF J3 Mux and Scope J9 Digiscope LT Mux and Scope J30 Digiscope UT Quad PSH 1 Card Quad PSH 1 J1 Pulse Pileup Det TOF J2 Quad PSH 1 J2 ADC J1 Quad PSH 1 J4 ADC J2 Quad PSH 1 J6 ADC J3 Quad PSH 1 J8 ADC J4 All Units Quad PSH 2 J7 Pulse Pileup Det TOF J4 Quad PSH 2 J8 Pulse Pileup Det TOF J5 4 10 5 4 SERVICE AND REPAIR PROCEDURES ELECTRONIC ADJUSTMENTS CABLE AND JUMPER CONFIGU
302. e Sorting Options speed e Use the XY adjustment to change the speed that the mechanism moves front to back and left to right Use 1 and 4 to increase or decrease the speed as needed 3 Use the Rotation adjustment to change the speed of rotation of the plate holder 4 Press Calibrate and observe the mechanism closely after any speed change Note Speed adjustments ensure that the mechanism operates smoothly without binding Operation at too high a speed results in faults during the Calibration cycle Too high a rotational speed results in the plate holder over rotating causing a fault condition The new speed values are retained by the system until Cytometer software is reloaded Note the numeric speed values for future reference Testing the Autoclone Sorting Option Mechanism You can perform two tests to test the mechanical and electronic systems In the first test the mechanism simply steps through each well position This provides test of the movement mechanism The second test generates sort pulses based on histograms acquired using FL test as the signal source This test provides further confidence in sorting electronics operation See Chapter 6 of the Options manual for additional information on adjustments Prepare the System 1 At the Cytometer screen a Select Options b Set DRIVE to 100 PN 4237003C 3 10 7 INSTALLATION PROCEDURES AUTOCLONE SORTING OPTION INSTALLATION Set FREQ to 0
303. e beams remain parallel through the flow cell all the way to the FALS sensor Note Lower laser power is easiest on the eyes It is recommended to place it right after the beam shaper and just before the scatter sensor PN 4237003C 3 12 7 INSTALLATION PROCEDURES BEAM TRANSLATOR UPGRADE 3 12 8 PN 4237003 INSTALLATION PROCEDURES 3 SORT PERFORMANCE ESP UPGRADE 3 13 SORT PERFORMANCE ESP UPGRADE Purpose Do this procedure to bring existing instruments to the correct revision level and to install the sort enhancement kit Do this procedure only on systems when installing the Sort Upgrade ESP kit Tools Supplies Needed Sort Upgrade Kit e PN 6913253 for Elite Systems without switch amps before SN U07122 and for Elite Systems with the Gated Amp option This kit includes switch amps e PN 6913250 for Elite Systems that have Switch Amps after SN 707122 and for Elite Systems with the Gated Amp option This kit does not include switch amps Sort Delay R3 card PN 6706034 Sort Oscillator R2 card PN 6705694 Pulse Pileup Det TOF card 6705954 Sort bracket PN 6856321 Control panel PN 6858680 UUUUU Preliminary Procedure l Verify that the kit is correct and complete 2 Verify that The software is version 4 0 or higher b Watchdog Timing has been done c Pneumatic Reliability Improvement Heading 4 5 PNEUMATICS SYSTEM has been done for low bleed regulators d The instrument is equipped with a 10
304. e chosen drop then there are less than 90 beads in the chosen drop Repeat steps 11 through 13 Note Your adjustment depends upon which direction the beads fell For example If the flow cell was raised too much the beads fell in the drop 28 before the chosen drop 29 if the flow cell was too low the beads fell in the drop after 30 the chosen drop 29 If the flow cell was turned the wrong way then most of the beads fell into the drop after 30 the chosen drop 29 Reduce the number stored to 20 Do a sort matrix on the phase of that droplet which would be 0 0 to 0 9 on the delay Choose one of the phases and ensure that the side streams are good Note Several phases should have 20 in them Sort several drops of 20 particles onto a slide and use the fluorescent microscope to count so that you are absolutely sure of your delay and phase Make the camera delay match the actual delay by using the Flow Factor adjustment option on the Camera screen Note The system stores this value unless the software is reloaded at which point it will have to be adjusted again for that particular flow cell PN 4237003C INSTALLATION PROCEDURES 3 SYSTEM TESTING Monitoring the Sort l Zoomin on the last attached droplet so that both the droplet position and connection neck are clearly visible 2 Monitor stream stability a Place one cursor tangent to the bottom of the last attached drop b Place the other cursor tangent to the bo
305. e delay cards For each channel the card uses the returned delayed signal or the undelayed signal Gates by switching the signal outputs between the signal inputs or ground The gate pulse from the Gated Amp Control card controls the gating Inputs The Scat Aux Gated Amp card has four signal inputs FALS PMT1 90LS Ausl and Aux2 PN 4237003C PN 42370036 INSTRUMENT DESCRIPTION 2 CARD DESCRIPTION GATED AMPLIFIER CONFIGURATION Figure 2 12 2 Block Diagram Scat Aux Gated Amp Card FAST 7 MICROSECONDS SLOW 20 40 60 MICROSECONDS DELAY CARD 2 DELAY CARD 2 This is a quad 4 channel This is a quad 4 channel card Other connections card Other connections are similar but not J8 shown for clarity TO DELAY CARDS 14 J15 This analog switch chooses between fast and slow delay are similar but not shown for clarity Note that all channels are set the same This switch chooses delayed or undelayed these are individually controllable J4 Le Aux1 DCRESTORE J20 AUX 1 OUT IN and 10x GAIN E Sy ire m d e scel x This switch allows the signal to pass or 1 grounds the output J9 J17J16 1 1 1 1 eo i J5 auo DCRESTORE he i n and 10x GAIN Cho a Teg d AUK 2 g
306. e optical bench is referred to as the columns The shorter dimension of the optical bench is referred to as the rows 1 Mount the UV laser in the farthest rear position possible on the table top with room for adjustment of the laser X Y axis column 16 row 9 Note You may want to mount the HeCd lasers closer to the flow cell 2 Parallel with the long dimension of the optical bench leave room in front of the laser output for the beam expander reducer a target and the optical rail 3 Mount the Argon laser in the column 8 9 and 10 row 7 Note Leave room in front of the laser output for the beam expander reducer a target and the optical rail 4 Mount the HeNe laser in its usual position next to the fluorescence collection assembly Note Leave room for alignment of the X Y axis of the HeNe laser between it and the fluorescent collection assembly Leave room for a target a beam expander reducer and the optical rail Laser Alignment for Gated Amp Systems Standard Laser Alignment l Decide which laser needs to be used as a trigger source for the Gated Amp Note This needs to be the laser the cell hits first 2 Decide which lasers are to be used and in what time space dimension top beam or bottom beam Note For this alignment procedure we will align the Argon on top and the HeNe and UV lasers on the bottom 3 Install the two standard 5 in targets on the optical bench in front of the HeNe laser separated
307. e optical plate 12 Remove the six bolts located in step 11 a Note the location of the shorter bolt Do not allow optical plate to fall b Carefully set aside optical plate PN 4237003C 13 INSTALLATION PROCEDURES COLLECTION OPTICS UPGRADE Remove the pickup lens assembly by removing the four bolts securing the pickup lens assembly to the PMT housing Replacement Procedure 1 Mv eoo qui 10 11 PN 4237003C Attach the replacement pickup lens to the PMT housing with the four bolts removed in step 13 of the Removal Procedure Reinstall the optical plate using the 6 bolts removed in step 12 of the Removal Procedure Align plate with mark made in step 10 of the Removal Procedure Tighten bolts slightly Remove laser block you installed in step 7 of the Removal Procedure allowing the beam to strike the target s on the optical plate Verify the laser beam passes cleanly through the target apertures a Loosen the plate mounting bolts enough to shift the plate position until the laser beam passes cleanly through the target apertures b Tighten the bolts securely and check the that the beam passes through the targets again c Repeat step a and b until proper alignment is achieved again Note If you used a power meter as suggested before step 6 of the Removal Procedure ensure that the laser power past the apertures is the same or better than before Reinstall the a FALS detector b Beam shutter c Flo
308. e pressure more than 0 20 psi above optimal setting for running beads Avoid compromising sort stability do not increase the sample pressure more than 0 20 psi above optimal setting for running beads 12 Run the sample by either of the following methods Run the actual sample at the data rate desired for sorting Note the sample pressure required to achieve the desired sort rate and use this same sample pressure when doing the sort matrix which means checking the Delay setting OR Run fluorospheres at the same concentration as the sample to be sorted 13 Examine the side streams while the sample is running 14 Use a 3 drop sort to ensure that the side streams are as clean as possible a make the streams cleaner you may need to adjust the Front Porch and Back Porch adjustments 1 ForESP systems the Front Porch and Back Porch adjustments are located on the front of the right pedestal panel as shown in Figure 3 6 2 2 Onnon ESP systems the Front and Back Porch adjustments are located on the Sort Oscillator card e Adjust R93 for the best side streams while keeping the front porch level between 6096 and 9596 of the main pulse e Adjust R92 for the best center waste stream while keeping the back porch level between 596 and 3096 of the main pulse PN 4237003C PN 42370036 INSTALLATION PROCEDURES SYSTEM TESTING b Achange in sheath pressure may require you to readjust the front and back porch even if you d
309. e second cell if the followed by a to sort both cells second cell falls in another droplet adjacent to the first cell s sort good cell window more than four quarter droplets away This means that In a one droplet sort if the second cell falls in the next drop the both cells meet sort window extends to form a two droplet sort A three droplet the criteria to sort can extend to five droplets sort in the same direction Note Extension occurs under the above conditions in any sorting coincidence mode including Abort OFF Extension is actually an unabort Note With Abort ON in any mode extension will not occur if the cells are close enough together to fall in the same droplet which means within four quarter droplets from one another In this case the system aborts both cells Good cell Truncation System sorts the The system truncates the pulse if followed by a good cell but The system is performing a sort of at least two droplets and bad cell the bad cell This means that a cell does not meet all the criteria to sort in the same direction as the first cell The bad cell occurs in one of the droplets in the first cell s sort window and Abort is ON any mode except Abort OFF Definitions Good a particle or cell that meets the conditions to be sorted meets the sort equation by being in the correct regions and or bitmaps Bad a particle or cell that does not meet the conditi
310. e slot Wet deflection plates Remove the plates Clean with deionized water Dry thoroughly and replace Bubble in the flow cell Press CLEAR several times until wavering stops Sample tubing needs to be changed Change the sample tubing No deflection No deflection of side streams when Sort Test is on and deflection is approximately 85 Deflection plates arced and shorted out 1 Press RESET on the Cytometer If that does not work power down the system then power up again 2 Clean and dry plates before replacing Blown fuse on Sort Output card Replace the fuse 7 5 4 PN 42370030 Table 7 5 1 Troubleshooting Sort Problems Continued TROUBLESHOOTING TROUBLESHOOTING THE SORT SUBSYSTEM Condition Symptom Problem Corrective Action Run of sorted samples is 9096 Low sort purity Coincidence and or PPU not used in sort setup Enable Coincidence Abort and PPU Sort tubes too close to center stream Increase deflection 5 1096 and move sort tubes out slightly Sort regions include unwanted cells not seen on histogram being displayed Include all histogram gates in the sort equation Discriminator set too high Lower the discriminator setting Sort delay and phase not set properly Review sort setup procedure and ensure the correct drop delay is sorted Too many drops sorted Use a 1 drop sort window So
311. e the flow cell stage to the vertical plate only enough to allow the flow cell to be tilted b Adjust the flow cell tilt as needed c Tighten the bolts Center the flow channel in the laser beam using the adjustment on the Piezo assembly Observe the reflected laser beam and path of the beam through the flow cell Repeat the above adjustments to ensure a The laser beam reflected from the flow cell is coincident with the incoming laser beam b The laser beam is centered on the flow channel PN 4237003C INSTALLATION PROCEDURES WATCHDOG TIMING 3 3 17 WATCHDOG TIMING Purpose This procedure extends the watchdog time out and prevents momentary dropouts in 24 V power which can occur when the Cytometer CPU is busy Do this procedure when Elite Software Version 4 0 is installed Tools Supplies Needed 221 kOhm resistor PN 4717745 6 22 uF capacitor PN 1515017 3 Procedure 1 Remove the Sensor Interface card from the instrument Locate R77 and C44 located about 4 in 10 cm back from J9 between U21 and U22 Replace C44 with the 22 pf capacitor Remove the jumper from E25 E26 Reinstall the Sensor Interface card Do Heading 5 1 VERIFICATION INSPECTION PROCEDURE to ensure the instruments fluidics are normal o wem PN 4237003C 3 17 1 INSTALLATION PROCEDURES WATCHDOG TIMING 3 17 2 PN 4237003C INSTALLATION PROCEDURES 3 SOFTWARE VERSION 4 0 INSTALLATION 3 18 SOFTWARE VERSION 4 0 INSTALLA
312. e you do this procedure PN 4237003C INSTALLATION PROCEDURES SOFTWARE VERSION 4 01 INSTALLATION WARNING Risk of personal injury Wear barrier protection including but not limited to protective eyewear gloves and suitable laboratory attire when performing this procedure 1 Remove the waste bottle from the instrument a Disconnect the color coded connectors b Disconnect the level detector wires at the bracket Carefully remove the waste bottle cap from the waste bottle Look inside the clear plastic float to verify that the two magnets grayish metallic cylinders are attached to the bottom of the float See Figure 3 19 1 Figure 3 19 1 Waste Bottle Caps FOR 4 0 AND EARLIER FOR 4 01 AND LATER SOFTWARE VERSIONS SOFTWARE VERSIONS ITEM 1 FLOAT ITEM 2 RETAINING WASHER 7003180A 4 Carefully remove the plastic retaining washer See Figure 3 19 1 The washer has a slit to facilitate removal e Apply sufficient force to remove the washer being careful not to break it An extra washer is provided in case of breakage e Do not allow the float to fall to the table or floor Carefully slide the float off the waste bottle cap Rotate the float 180 degrees and slide it back onto the waste bottle cap Verify that the magnets are now attached to the top of the float PN 4237003C 3 19 3 INSTALLATION PROCEDURES SOFTWARE VERSION 4 01 INSTALLATION Carefully reattach the retaining washer Reconnect
313. ece of tubing will be needed Connect the tubing that was going to top of the removed T fitting to FF33 Note longer piece of tubing will be needed Do not remove check valve Remove tubing from the bottom of the T fitting coming from FF36 Add 1 inch 3 cm of the tubing from the bottom of the T fitting coming from FF36 Install the top of the new T fitting to the 1 inch piece of tubing Connect the tubing that you removed in step 24 to the bottom of the new t fitting Install the 1 inch 3 cm piece of the tubing on the remaining end of the new T fitting Install the T fitting on the 1 inch 3 cm piece of tubing just installed Connect the remaining ends of T fitting to FF39 and FF42 with tubing Remove the two screws holding the terminal block assembly mounted to the frame Pull terminal block out to where you can access the terminals Connect the spade lugs from the cable assembly to the terminals according to Table 3 10 1 PN 4237003C PN 4237003C Table 3 10 1 Spade Lug Connections INSTALLATION PROCEDURES AUTOCLONE SORTING OPTION INSTALLATION From Wire Marker WM Number To Terminal Block Label None STROBE None STROBE None DEFL None DEFL None 12V None 15V 433 Orange 15V 432 Wht BIk ANLG GND None ANLG GND 434 Green 15V None 15V 438 Black DGT GND None DGT GND None DGT GND None 5 None 5 437 Yellow 45
314. echanism Adjustment 2 2 4 OPTICAL COMPONENTS 2 3 1 Lasers 2 3 1 Beam Shapers 2 3 5 Beam Steering Optics 2 3 7 Beam Translators 2 3 8 Beam Expander Reducer 2 3 8 Side Scatter and Fluorescence Collection Optics 2 3 9 Sort Sense Flow Cell Tips 2 3 9 Jet in Air Flow Cell Tips 2 3 10 Fast Quartz Flow Cell 2 3 11 Optical Filters 2 3 11 Long Pass LP Filters 2 3 12 Short Pass SP Filters 2 3 12 Blocking BK Filters 2 3 12 Band Pass BP Filters 2 3 12 Dichroic Long Pass DL Filters 2 3 12 Dichroic Short Pass DS Filters 2 3 13 Neutral Density ND Filters 2 3 13 Forward Scatter Detector 2 3 13 Forward Scatter Obscuration Templates 2 3 13 ELECTRONICS SUBSYSTEM 2 4 1 Description 2 4 1 CONTROL AND DISPLAY SUBSYSTEMS 2 5 1 Multibus Subsystem 2 5 1 Cytometer Computer Subsystem 2 5 1 Multibus CPU 2 5 1 External Memory or 256K Memory Card 2 5 2 Serial I O Card 2 5 2 Laser Control Subsystem 2 5 2 Dual Laser Control Card 2 5 2 Video Subsystem Cytometer 2 5 4 Camera Image Signal Flow Video Display 2 5 5 Camera Interface Card 2 5 5 Dual CRT Control Card 2 5 5 Digiscope Card 2 5 5 Pulse Video Display 2 5 5 PN 4237003C 2 6 Lg 2 8 PN 4237003C CONTENTS Cytometer Control Subsystem 2 5 6 Description 2 5 6 Data Taker Interface Card 2 5 6 Gated Amp Control Card 2 5 6 Sensor I O Card 2 5 8 Motor Control Card 2 5 8 DAC Card 2 5 8 ACQUISITION SUBSYSTEM 2 6 1 Analog Signal Processi
315. ects the UV to the UV reflector which directs the beam through the beam expander reducer The expander reducer can be adjusted to make the UV beam spot size compatible with the 488 nm beam spot size The 488 nm beam passes through the attenuator wheel The wheel is a variable density filter that can be set to give from 0 to ND2 attenuation divide the 488 beam power by a factor between 0 100 Both beams pass through the translators These translators can be easily adjusted to independently shift the height of the two laser beams without affecting their angular alignment Tools Supplies Needed Enterprise laser kit Enterprise laser customer manual PN 7232520 6 Note Installation of this laser is identical to installation of the 1305 laser except for the laser WARNING Risk of personal injury You must be certified as trained on the Coherent 190 160 1300 lasers and you must be in contact with a technical support person who is trained on the Enterprise laser in order for you to qualify as an authorized installer If and only if you meet these requirements read the manual supplied with the laser especially Chapter 3 Utility and Environmental Requirements and System Installation and Chapter 6 Rear Mirror Adjustment Call your technical support person before continuing WARNING Risk of personal injury The Enterprise laser always emits UV as well as visible radiation Take appropriate precautions to avoid exposur
316. ed drops under the fluorescent microscope e Ar least 9096 of the beads should be in one drop If so select that drop and go to step 13 e 111655 than 9096 of the beads are in one drop go to step 9 Select the drop that contains most of the beads For example If most of the beads are in drop 29 and some are in 30 then select 29 to be the drop of choice Place a cursor tangent to the tip of the drop use the monitor to see the drop and be sure that the camera view is zoomed in sufficiently to easily view the last drop and the filament connecting it to the stream ATTENTION Make sure that the fluorescence signals do not drop when you raise the flow cell 11 12 13 14 15 16 17 18 Turn the flow cell vertical adjustment to raise the flow cell and stream 1 16 of a drop This makes the beads fall in the chosen drop 29 in the example Note This adjustment is more sensitive with quartz flow cell tips than with jet in air Sort the 100 beads using one delay before the chosen drop the actual delay of the chosen drop and one delay after the chosen drop Note Using the example in step 9 above this means that you would choose 28 29 and 30 Check the drops on either side of your chosen drop If there are 10 beads or less in the drops on either side of the chosen drop then there are 90 beads 9096 in the chosen drop Go to step 14 are 11 beads or more in the drops on either side of th
317. eded 4 10 1 Cable Configurations 4 10 1 Back Panel Wiring 4 10 1 Jumper Configurations 4 10 1 Multibus Card Cage 4 10 1 Non Gated Amp Card Cage 4 10 2 Gated Amp Card Cage 4 10 2 Data Acquisition Card Cage Configuration 4 10 3 Analyzer HV DAC Control Card 4 10 4 PC Model 486 4 10 4 Cable Connections 4 10 4 Procedure 4 10 4 SORT SENSE FLOW CELL ADJUSTMENT AND VERIFICATION 4 11 1 Purpose 4 11 1 Tools Supplies Needed 4 11 1 Procedure 4 11 1 Replacing Metal Fittings with PVC Fittings 4 11 2 Flow Cell Replacement and Verification 4 11 4 4237003C PN 42370036 4 12 OPTICAL ALIGNMENT PROCEDURE 4 12 1 3 13 Purpose 4 12 1 Tools Supplies Needed 4 12 1 Flow Cell Alignment 4 12 1 Parabolic Mirror on Tip Optional 4 12 4 Laser Beam Alignment 4 12 5 Optical Fine Tuning 4 12 8 PMT4 System 4 12 9 PMT5 System 4 12 11 EXTERNAL MEMORY CARD REPLACEMENT 4 13 1 Purpose 4 13 1 Tools Supplies Needed 4 13 1 Procedure 4 13 1 5 MAINTENANCE PROCEDURES 5 1 1 341 3 2 VERIFICATION INSPECTION PROCEDURE 5 1 1 Verifying Instrument Performance Every Call 5 1 1 Cytometer 5 1 1 Workstation 5 1 1 System Not Serviced Within Past 30 Days 5 1 1 Operational Test Cytometer Electronics Optics or Fluidics 5 1 1 Operational Test Sorting Ability 5 1 2 Operational Test Every Call 5 1 2 TOF Verification 5 1 2 Purpose 5 1 2 Procedure 5 1 2 PREVENTIVE MAINTENANCE PMI PROCEDURE 5
318. een is in increments from 0 to 400 When the Cytometer Software is downloaded the system resets and the Cytometer Touch Screen displays the Main Control screen PN 4237003C 4 9 3 SERVICE AND REPAIR PROCEDURES SOFTWARE INSTALLATION 4 9 4 PN 4237003C SERVICE AND REPAIR PROCEDURES ELECTRONIC ADJUSTMENTS CABLE AND JUMPER CONFIGURATIONS 4 10 ELECTRONIC ADJUSTMENTS CABLE AND JUMPER CONFIGURATIONS Purpose Too This section provides the necessary procedures for you to make adjustments to the Elite s electronics For information on Gated Amplifier cards refer to Heading 3 9 GATED AMPLIFIER UPGRADE For information on Sorting Electronics refer to Heading 3 13 SORT PERFORMANCE ESP UPGRADE Is Supplies Needed None Cable Configurations Back Panel Wiring Verify the back panel wiring is configured as shown in the Table 4 10 1 Table 4 10 1 Back Panel Wiring Configuration Top AC Receptacle 16 Bottom AC Receptacle 17 System Power Laser Power WMT71 black WM74 black WM72 white WMY73 white G earth gnd E13 G earth gnd E14 Jumper Configurations Multibus Card Cage Jumper the cards in the Multibus card cage as shown in Table 4 10 2 Table 4 10 2 Multibus Card Cage Cards Jumper and Switch Settings Card Jumper and or Switch Settings CPU IBC 86C Intel factory configured Serial 1 0 None Data Taker Interface None PN 4237003C 4 10 1 4 SERVICE AND REPAIR PR
319. eft Front Corner Inside View See Figure 8 2 16 8 2 16 Analyzer Sample Drawer Detail of Parts See Figure 8 2 17 8 2 17 Analyzer Top Inside View Detail of Parts See Figure 8 2 18 8 2 18 Elite Flow Cell Area Detail of Parts See Figure 8 2 19 8 2 19 Elite Analyzer Flow Cell Area Detail of Parts See Figure 8 2 20 8 2 20 Deflection Body Detail of Parts See Figure 8 2 21 8 2 21 Photomultiplier Tubes PMTs Detail of Parts See Figure 8 2 22 8 2 22 Autoclone Sorting Option Mechanism Detail of Parts See Figure 8 2 23 8 2 23 PN 4237003C 8 2 24 8 2 25 8 2 26 8 2 27 8 2 28 8 2 29 8 2 30 8 2 31 8 2 32 8 2 33 8 2 34 8 2 35 8 2 36 8 2 37 8 2 38 1 1 2 1 2 2 2 3 2 4 2 5 3 1 PN 42370036 Autoclone Sorting Option Beak Waste Catcher Detail of Parts See Figure 8 2 24 8 2 24 Autoclone Sorting Option Location and Connections Detail of Parts See Figure 8 2 25 8 2 25 Autoclone Sorting Option Accessories Detail of Parts See Figure 8 2 26 8 2 26 Transformer Drawer Assembly Detail of Parts See Figure 8 2 27 8 2 27 Computer Box Assembly Detail of Parts See Figure 8 2 28 8 2 28 Beam Splitter Dichroic Detail of Parts See Figure 8 2 29 8 2 29 Beam Shaper Detail of Parts See Figure 8 2 30 8 2 30 Elite ESP Flow Cell Stage Detail of Parts See Figure 8 2 31 8 2 31 Elite Flow Cell Stage Detail of Parts See Figure 8 2 32 8 2 32 Model 74 HeCd Laser
320. eft in the top cage 20 Install the new Dual FL SW R Card in third slot from left in the top cage and jumper as 1 E1 E2 Out 21 Install the new 3 PMT Sub SW R card a 4 PMT Jumper as 1 1 2 OUT E3 E4 OUT E5 E6 OUT and install in fourth slot from left in the top cage b 5PMT Jumper as 2 1 2 OUT E3 E4 IN E5 E6 OUT and install in fourth slot from left in the top cage PN 4237003C 3 9 3 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE 22 See Table 3 9 1 and jumper a The new 3 Sub SW R card 1 E1 E2 OUT E3 E4 OUT E5 E6 OUT and install in fifth slot from left in the top cage The new Gated Amp card as 1 E1 E2 OUT and install in sixth slot from The new Quad 20 40 60 Microsecond Delay card as 2 E1 E2 IN E3 E4 IN The new Quad 7 Microsecond Delay card as 2 E10 E15 and install in 8th slot The new Quad 7 Microsecond Delay card as 1 E10 E11 IN and install in 9th slot The new Quad 20 40 60 Microsecond Delay card as 41 E1 E2 IN E3 E4 IN The new Scat Aux Gated Amp card as 1 E2 E3 OUT and install in 11th slot The new 3 PMT Sub SW R card as 3 1 2 IN E3 E4 OUT E5 E6 IN and install The new Peak Scatter Mux SW R card as 1 2 IN E3 E4 IN and install in 13th The new Scat CV SW R card as 1 E1 E2 OUT E3 E4 OUT and install in 14th slot b left in the top cage E10 E11 IN and install in seventh slot from left top cage d from left in the top cage
321. egulator is adjusted correctly place your DVM lead on TP4 Read the liquid pressure sensor output The voltage here depends on the sheath liquid pressure Note You will read about 10 V at 15 psi and 8 V at 12 psi or segun guid pressure x 10Volts Volts The sheath liquid pressure is the same as the sheath air pressure only if there is no liquid in the system or the sheath tank level is the same as the sensor level this is true when the bottle is half full Since 1 psi 27 68 in of water and the liquid level in the bottle differs by 7 inches from empty to full the liquid pressure at the sensor will be 0 25 psi higher when the bottle is full than when it is empty Fill the sheath bottle halfway with sheath fluid connect your meter toTP4 and adjust the voltages a Set Sheath Pressure to 12 psi You should read 8 00 V on your meter If not adjust R75 for the correct voltage b Set Sheath Pressure to 6 psi Wait for the pressures to stabilize then verify that you meter reads 4 00 V If not adjust R76 as needed c Repeat steps a and b until you read the correct voltage for both pressures PN 4237003C TROUBLESHOOTING TROUBLESHOOTING PRESSURE CONTROL PROBLEMS 7 Note If the correct voltages cannot be obtained in this step the in line sheath pressure sensor or the Pneumatic Interface card may be defective 9 If the voltage is correct but the display on the Cytometer screen is not the problem is the Sensor Interface
322. elect PMT 2 Peak for Trace 1 b Select PMT 5 Peak for Trace 2 c Change 2 3 4 5 Int Gain to 30 d Change PMT 2 3 4 5 Peak Gain to 30 e Increase PMT 2 HV for a 3 Division Tall Peak Pulse on Trace 1 f Increase PMT 5 HV for a 3 Division Tall Peak Pulse on Trace 2 8 At the Scope Screen Select PMT 2 Peak for Trace 1 b Select 5 Peak for Trace 2 c Change PMT 2 and PMT 5 Peak Gains all around and ensure the amplitudes follow as expected Put Gain back to 30 9 At the Scope Screen a Select PMT 3 Peak for Trace 1 b Select PMT 4 Peak for Trace 2 c Increase PMT 3 HV for a 3 Division Tall Peak on Trace 1 10 Insert the 575 BP filter in the slot under PMT 4 11 Increase PMT 4 HV for a 3 Division Tall Peak on Trace 2 12 Change PMT 3 Peak and PMT 4 Peak Gains all around and ensure the amplitudes follows as expected Put Gains back to 30 13 At the Scope Screen a Select PMT 2 for Trace 1 b Select PMT 3 Int for Trace 2 4237003C PN 42370036 INSTALLATION PROCEDURES 5TH PMT KIT WITH SWITCHABLE AMPS INSTALLATION PROCEDURE 14 At the FL Compensation Screen 15 16 1T 18 19 20 a Select PMT 2 minus PMT 3 Int for 5096 and ensure Trace 1 decreases by 5096 Put back to 096 Select PMT 2 minus PMT 4 for 5096 and ensure Trace 1 decreases by 5096 Put back to 096 Select PMT 3 minus PMT 2 for 5096 and ensure Trace 2 decreases by 5096 Put back to 096 Select PMT 3 minus and PMT
323. ement theory Table 2 3 3 shows approximate working beam spot dimensions obtained using various combinations of laser and focusing lens To use the table Locate the appropriate laser and wavelength in the table Read the beam height and width from the appropriate columns for the focal length of the lenses in the beam shaper you are using For example Coherent 305 Laser with 488 nm using 8 x 80 defocused 5 8 mm beam shaper gives the following beam height of 3 4 um and beam width of 112 um Table 2 3 3 Working Beam Spot Dimensions Laser Wavelength Beam Front Lens Focal Length mm Rear Lens Focal Length mm and nm Diameter Defocus Distance mm 8 10 15 40 60 80 80 80 5 8 15 1 Beam Height um Beam Width um Cyonics 488 0 67 6 4 8 0 12 1 334 51 3 700 869 151 Uniphase 633 0 68 7 6 9 5 144 1400 619 850 102 169 Melles Griot 544 0 80 6 3 7 8 11 8 321 488 670 898 172 DPSS 532 0 60 8 0 100 1152 424 658 907 107 175 Omnichrome 325 1 30 4 1 5 1 7 7 206 309 412 137 343 Coherent 300 3514365 1 24 3 6 4 5 6 8 18 2 274 366 169 432 Series and Coherent 457 1 42 3 3 4 1 6 1 16 3 245 32 7 108 271 Spectrum 488 1 46 34 42 63 169 254 339 112 288 515 1 50 3 5 4 3 6 5 174 126 1 349 114 286 Spectrum only 647 1 50 6 3 7 9 118 131 6 147 4 633 1171 418 Coherent 3514365 0 67 3 4 4 2 6 4 17 4 266 36 1 81 5 1193 Enterprise 488 0 80 3 7 4 6 7 0 19 0 29 0 394 1105 256
324. ements 3 13 2 cycle defined 2 8 1 droplet when it can be sorted 2 8 8 drops microscopic viewing of 3 6 6 illustration of filament connecting last drop 4 11 6 installation procedures 3 13 3 installing the ESP upgrade 3 13 1 matrix 3 6 6 monitoring the sort 3 6 7 operational test See VIP porch settings 3 6 5 pulse amplitude adjustment 4 2 3 pulse amplitude test procedure 4 2 2 A 3 1 quarter drop sort defined 2 8 8 stream stability 3 6 7 temperature affect of 3 1 3 tube placement 3 6 2 verifying pulse pileup 4 2 11 verifying the sort 4 2 4 sort function verification adjusting the phase 4 2 5 adjusting the pulse generator calculating the delay 4 2 7 checking the sort 3 6 1 monitoring the sort 4 2 8 pulse generator setup 4 2 10 INDEX 15 INDEX setting the drive frequency sort settings 4 2 4 setting up for sorting 4 2 10 testing for extended abort 4 2 13 testing for sort pulse truncation 4 2 12 verifying sort coincidence 4 2 9 verifying sort pulse extension 4 2 12 Sort Oscillator card block diagram 2 8 10 coincidence logic function 2 8 10 function 2 8 10 possible coincident conditions 2 8 12 stops the sort when 2 8 7 verifying sort coincidence after replacing the card 4 2 9 Sort Oscillator R2 card jumpers 4 10 3 A 2 4 Sort Output card adjusting for sort pulse amplitude 4 2 3 block diagram 2 8 14 crystal drive adjustments 4 2 2 A 3 1 function 2 8 13 4 10 4 jumpers 4 10 4 A 2
325. en the HeNe laser output shutter and run fluorospheres at a data rate of approximately 100 Align the optical area for maximum signal on both traces of Digiscope using the standard approach a Flow cell X Y adjustment b Beam shaper X Y Z adjustment c Fluorescence collection assembly focus snout Align the Argon laser Open the Argon laser output shutter b Align for maximum collinear signals both Argon and HeNe signals appear to be on top of each other on both traces of Digiscope using Argon laser 90 degree dichroic adjustments c Close the Argon laser output shutter 42370036 SERVICE AND REPAIR PROCEDURES 4 OPTICAL ALIGNMENT PROCEDURE ATTENTION Do not change any flow cell or beam shaper alignment because you will undo the alignment that you just did for the HeNe 10 Align the UV laser a Open the UV laser output shutter b Align for maximum collinear signals on both traces of the Digiscope using the 90 degree mirror only ATTENTION Do not change any flow cell or beam shaper alignment because you will undo the alignment that you just did for the HeNe 11 Open the Argon laser output shutter 12 Now that all three lasers are active verify that all three resultant signals occur in the same time frame collinear all three appearing to be on top of each other a Block each laser off one at a time with a target an opaque object b Observe the change in displayed signals on the Digiscope Gated A
326. eplace the flow cell Move the camera all the way down stream and look at the droplets to verify they are clear sharp and fairly round in shape without any satellite droplets Ifthe droplets are acceptable go to step 8 Ifthe droplets are not acceptable slightly adjust the frequency using the decimal setting to improve the shape and clarity of the drops Move the camera back upstream and ensure that the last attached droplet position has not changed significantly by more than 3 drops Note You may need to adjust the sort deflection body slightly to view the last attached drop It is helpful if it is visible just above the ground plate so that it can be easily monitored Phase Adjustment M E m Set Sort Counters to OFE Turn on the Sort Test Turn on the Left Sort and Right Sort Turn on the High Voltage Set Sort Droplet to 1 and press START Sort counters should be running Increase the deflection by increasing the Deflection amplitude to bring out the side streams enough but do not allow them to hit the sort plates about 9096 on the standard sort plates Use the phase adjustment the decimal portion of the delay setting to find the best side streams For the best side streams a Find the worst phase setting and subtract 0 5 b Setthe phase to that number for the best side streams You should get at least seven of the phase settings to give good clean side streams If not begin again at step 1 4
327. er Collinear Laser Alignment Fine Tuning Procedure l Qv MU mo 10 11 12 13 Remove the two 5 in 13 cm optical bench mounted laser alignment targets and the two vertical optical plate mounted targets Install the scatter sensor Install the beam shaper Install the laser shutter Use the flow cell vertical adjustment knob to lower the flow cell to its proper position Verify the proper position of the flow cell by using the camera to view placement Note Remove the 525 BP filter to obtain a side scatter fluorescent signal in PMT2 Except for the 1 0 ND filter remove all optical filters from the forward scatter sensor filter holder Install the filter holder back in the FALS Sensor Press SHEATH to initiate sheath flow Obtain traces on Cytometer s Scope screen a Choose Fals Peak for lower trace b Choose PMT 2 Peak for upper trace Open HeNe laser output shutter Run DNA Check Beads at a data rate of approximately 100 Align optical area for maximum signal on both traces of Digiscope a Flow cell X Y Adj and Rotation b Beam shaper X Y Z Adj c Fluorescence collection assembly focus snout 4237003C PN 42370036 14 INSTALLATION PROCEDURES BEAM TRANSLATOR UPGRADE Open the Argon laser output shutter ATTENTION Do not change any flow cell or beam shaper alignment otherwise you will undo the alignment that was just completed for the HeNe 15 16 1T Use the
328. er is still locked up clear the CPU s memory to restore normal operation Tools Supplies Needed None Reset the Cytometer CPU 1 Ifthe CPU locks up locate the RESET switch on the switch panel 2 Press and release the RESET switch to reset the Cytometer CPU 3 If proper functioning is restored resume normal operation Note If the Cytometer is still locked up the CPUS memory must be cleared to restore normal operation Follow the instructions under heading Clear the CPU s Memory on the Cytometer as written Clear the CPU s Memory on the Cytometer Note Clearing the CPU s memory allows an operator to easily reprogram the memory on the Cytometer without having to wipe out the memory from the Service screen Locate the RESET switch on the switch panel 2 Press and hold the RESET switch for approximately 10 seconds until a tone is heard from the External Memory card The External Memory card is located in the Multibus card cage 3 Atthe Workstation keyboard press F9 When the prompt to transfer the program into the Cytometer appears select Yes to initiate memory transfer from the Workstation to the CPU The number 1 appears on the Cytometer right monitor and begins to sequence as the files are transferred 5 Wait while the number displayed on the right monitor sequences or counts from 1 to approximately 425 This process takes several minutes 6 Resume normal operation 4237003 7 6 1 7
329. er of the printer To service a laser refer to the manual supplied by the manufacturer of the laser Any service memo that affects the information in this manual will include a Notice of Information Update form for this manual The Notice of Information Update form will summarize the changes and will list the specific headings figures and tables affected Intended Audience To use this manual effectively you need the following An operators knowledge of the Elite Service training on the Elite basic understanding of gt Cytometry terms and concepts Computer hardware The ability to gt Read pneumatic hydraulic schematics and understand related terminology gt Read electronic schematics and understand related terminology gt Use a digital volt meter DVM and an oscilloscope gt Use basic mechanical tools and understand related terminology PN 4237003 1 1 1 INTRODUCTION MANUAL DESCRIPTION Organization The material in this manual is organized into eight chapters and two appendices To make it easier to access the information Format In the online manual each page has a Contents button linked to a master table of contents and an Index button linked to an alphabetical index In the printed manual there is a master table of contents at the beginning of the manual a chapter specific table of contents at the beginning of each chapter and an alphabetical index at the end of the manual CHAPTER 1
330. ernoulli kit Switcher replacement kit Profile 11 Software Easy 2 2 Drive 20 MB Bernoulli field upgradable PN 4237003C 8 1 25 8 PARTS LISTS MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY Table 8 1 23 Workstation Continued Part Number Description Figure Item 6912816 6912866 6912881 6912970 6912973 6913112 6913113 6913114 6913115 6913117 6913149 6913170 6913258 6913259 6913343 6913404 6913407 6915005 6915006 6928258 7231244 7260019 Software Elite 4 0 with PrintQue FlowNet network kit Cartridge data 21 4 MB Drive optical multi function Switcher 12V Drive disk 90 MB Bernoulli removable Cartridge printer Laserjet 4 with Font Software Elite 4 01 Printer Laserjet 4 Printer Deskjet 500 Hewlett Packarde Labels 100 sheets 8 5 x 11 in Labels barcode generation kit Software 1 5 XL without cable Software 1 5 with serial cable Labels barcode 3 rolls Drive 150 MB Bernoulli ISA Drive disk Tahiti 11 without SCSI card Kit Pentium 90 MHz full workstation upgrade Kit Pentium 90 MHz upgrade Cable serial communication 9 25 pin Disk service pre final test Battery pack Intel 8 1 26 PN 4237003C PARTS LISTS 8 ILLUSTRATED PARTS 8 2 ILLUSTRATED PARTS Figure 8 2 1 Elite Cytometer Right Panel Removed See Table 8 2 1 E E q ele eeeeee _
331. ert the tall targets and target the Hehe and Argon lasers Remove the tall targets c Insert the short targets in place of the scatter sensor and the beam shutter and target the lasers beginning with the laser closest to the sampling compartment d Remove the short targets Remove the flow cell tip Place the HeNe mirror in front of the HeNe laser Place the 488 mirror in front of the Argon laser Use the mirror mount adjustments to align the two lasers through the targets a Loosen both the rail setscrew and the fine adjustment setscrew 3 7 5 INSTALLATION PROCEDURES OPTIONAL LASER INSTALLATION b Slide the mount to the approximate location desired c Tighten the rail setscrew just enough to keep the mount from wobbling but still allowing it to slide on the rail d Tighten the fine adjustment setscrew be careful not to overtighten e Use the fine adjustment knob to make the final location adjustment f Tighten the rail setscrew to lock the mount in place be careful not to overtighten WARNING Risk of personal injury The HeCd and water cooled Argon lasers operate in the UV ultraviolet mode which can injure you when exposed Use extreme care to avoid exposure Carefully note the probable beam paths before firing the laser and block the beam from exiting the instrument by using a piece of white paper such as a business card to locate the beam during alignment 15 Install the HeCd or water cooled Argo
332. es the sort stream onto the camera Many rays of light leave a point on an object However for clarity in describing image formation this example uses only two rays Figure 2 7 3 shows how the positive lens forms an image PN 4237003C INSTRUMENT DESCRIPTION 2 OPTICAL ELEMENTS Figure 2 7 3 Image Information La gt lt gt FOCAL FOCAL LENGTH LENGTH 7003172 In the example two light rays that leave a point on the object are traced through the lens The top ray is parallel to the optic axis and the lens refracts the ray to pass through the focal point The bottom ray leaves the same point on the object and passes through the front focal point of the lens Because this ray enters the lens from the focal point the lens refracts the ray to be parallel with the optic axis The lens forms one point of the image at the intersection of the top and bottom rays Note rays that leave the same point on the object and pass through the lens converge on the same image point Negative Lens A negative lens Figure 2 7 4 is a concave spherical surface and is opposite from the positive lens negative lens refracts rays of light so they diverge or spread out The point from which these rays appear to diverge is the focal point Figure 2 7 4 Negative Lens Focus FOCAL LENGTH 7003173A Equations There are two equations that apply to the lenses used in the Elite the Lensmakers equation and the Thin Len
333. ets in front of the UV laser WARNING Risk of personal injury from laser beam exposure Wear a pair of latex surgical gloves to reduce the amount of UV in the event you accidentally place your hands in front of the beam Remove all jewelry rings watches etc and be careful of tools that could reflect the beam 17 18 19 20 PM 22 23 Open the laser output shutter on the UV laser Align the UV laser using the Standard Laser Alignment procedure for two optical bench mounted 5 in targets then for the two smaller vertical optical plate mounted targets using the 90 degree full mirror Note Use a business card or fluorescent day glow paper to help detect position of the UV laser position Close the laser output shutter on the UV laser Place the screw on targets input and output lenses on the beam reducer Note The old style beam reducer can be used but it does not have built in targets Install the beam reducer on the optical bench as close as possible to the output of the UV laser Note Leave enough room to remove the screw on targets from the beam reducer when done Open the UV laser output shutter and align the X Y Z axis of the beam reducer for optimum alignment through the two optical bench mounted targets and the two vertical optical plate mounted targets Remove the screw on targets from the beam reducer when you are satisfied with the alignment of the beam reducer 4 SERVICE AND
334. ew See Table 8 2 13 77 77 1 7003112A Table 8 2 13 Cytometer Inside Rear View Detail of Parts See Figure 8 2 13 Item Part Number Description 1 6704327 36 V Regulator card 2 6856636 PMT power supply 3 6703950 HV DAC PMT backplane 4 6804637 PMT Bertan HV Supply card 5 6704679 Motor Controller card 6 6705191 Autoclone Sorting Option Controller card 7 6857276 Fiber optics light source 8 4004074 Switcher power supply 400W 9 3908025 Halogen lamp 10 6853275 15 V 2 A power supply 11 6856870 15 V 0 25A power supply 12 6854358 15 V 5 A power supply 13 6703941 PMT DAC R card 14 4004024 90 V power supply 15 6704960 Deflection power supply HV 3000 V PN 4237003C 8 2 13 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 14 Cytometer Right Rear Door Inside View See Table 8 2 14 7003099A Table 8 2 14 Cytometer Right Rear Door Inside View Detail of Parts See Figure 8 2 14 Item Part Number Description 1 6856866 Transformer rack rear door 2 2603065 Fan blower 112 CFM 115 Vac 3 2510014 Shock mount 4 6028002 Cable assembly ground 5 2105057 Power connector 6 6027967 Cable assembly ground 8 2 14 PN 4237003 PARTS LISTS 8 ILLUSTRATED PARTS Figure 8 2 15 Cytometer Left Rear Door Inside View See Table 8 2 15 7003100A Table 8 2 15 Cytometer Left Rear Door I
335. ews that hold the Compressor to the brackets on the side of the pneumatics compartment as shown in Figure 3 2 4 c Lower the Compressor to the floor d Position the Compressor so none of it touches the instrument Figure 3 2 4 Screws Securing Compressor to Pneumatics Cabinet REMOVE SCREWS 7003162A 9 Reinstall the rinse bottle holder 10 Remove the pinch valve deactivator clips Note There is a pinch valve deactivator clip located in the flow cell area as shown in Figure 3 2 5 PN 4237003C 3 2 5 INSTALLATION PROCEDURES INITIAL SETUP Figure 3 2 5 Pinch Valve Deactivator Clip One pinch valve deactivator clip located here m 7003002A 11 Install the filters a Open the filter compartment b Insert the filters from the filter kit into the appropriate slots See Figure 3 2 6 ATTENTION If installing an Elite Analyzer skip step 12 and go to Heading 3 3 Figure 3 2 6 Filter Slots 575BP 525BP 488BP 3 amp lt o S NO co Co Se L L 5904023 3 2 6 4237003 INSTALLATION PROCEDURES 3 INITIAL SETUP 12 Install the sample collection compartment Figure 3 2 7 components by installing a waste stream collector beak b Thecompartment floor c The sort tube holders Figure 3 2 7 Sample Collection Compartment WASTE STREAM COLLECTOR BEAK SORT TUBE HOLDERS COM
336. extra large 6027225 Cord 3 18 ga HeCd laser 8 2 38 6 8 1 18 PN 4237003C Table 8 1 14 Miscellaneous Continued MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY PARTS LISTS Part Number Description Figure Item 6214106 Valves check for 0 156 i d tubing waste line 8 2 6 2 11 6801536 6603488 DNA Check Beads kit 6703953 Detector pump motor analyzer 8 2 18 6 6856479 Filter coax ac 6856911 Drawer transformer 8 2 27 1 6857479 Computer box 3100 8 2 28 1 6858680 Rack Data Acquisition 6912798 Amplifier gated 8547008 Fluid sheath IsoFlow 3 liter Table 8 1 15 Optics Part Number Description Figure Item 3814233 Table optical 8 2 34 8 6604656 Beam shaper 15 x 80 mm confocal optional 6604657 Beam shaper 15 x 60 mm confocal 6604658 Beam shaper 15 x 80 mm A 15 6604659 Beam shaper 15x 80 mm A 6 6604660 Beam shaper 8 x 80 mm A 15 6604895 Beam shaper 10 x 80 mm A 15 optional 6856826 Beam shaper 8 x 80 mm Elite Analyzer 8 2 20 6857111 Beam shaper 15 x 80 mm Elite Analyzer 8 2 20 6857630 Beam shaper 40 x 80 confocal 6857634 Stage beam splitter 8 2 29 5 6858019 Beam shaper 15 x 80 mm Elite Analyzer 8 2 20 4 Table 8 1 16 Pneumatics Part Number Description Figure Item 1005697 Fitting hose barb metal small 0 062 i d 8 2 6 7 1020380 Bracket rinse 8 2 4 10 2306046 Transducer sheath pressure 8 2 6 8 2306051 Bracket mounting transducer 8 2 6 9 2603012 Blower
337. f integration capacitors to compensate for varying pulse widths One capacitor is for quartz flow cell operation low bandwidth and the other capacitor is for jet in air flow cell operation high bandwidth For all circuit cards with SWITCH or SW in their name the CPU sends a control code to the card that activates relays on the card This switches the correct capacitors into the appropriate circuits The older circuit cards in the Non Gated Amp systems do not have the SWITCH SW designator The jumpers on the card select the correct bandwidth 2 6 5 2 INSTRUMENT DESCRIPTION ACQUISITION SUBSYSTEM Figure 2 6 6 shows the typical Switch Amp Integrator circuit Figure 2 6 6 Typical Switch Amp Integrator Circuit 2200 pF 400 pF Peak 5K NAV 100 K signal AA in D Integral signal out p Gated Amp Concept 7003058A A Gated Amp separates fluorescent signals that are generated as a result of excitation by two different laser beams where the respective emission spectra overlap and the wavelength optical filters and subtraction are not effective Figure 2 6 7 shows the concept of the Gated Amp using a typical application as an example with a group of cells labeled with two different dyes Propidium Iodide PI and Hoechst Table 2 6 1 lists the emissions and requirements for each dye Table 2 6 1 Dye Emissions Dye Light Emitted Laser Needed Propidium lodide PI Range of w
338. f the right CRT An array of infrared emitting LEDs and phototransistors is mounted on the card They are aimed at each other to form a grid of horizontal and vertical light beams in front of the CRT face A circuit on the card scans the photo transistor outputs to detect when a pair of the horizontal and vertical beams has been blocked This information in encoded and sent to the Serial I O card to inform the Multibus CPU card where you have placed your finger on the screen The KIR3 card also receives signals from the membrane panel indicating which membrane button you pushed This information is also encoded and sent to the Serial I O card A dedicated microcomputer with RAM and ROM on the chip controls the operation of the KIR3 card The KIR3 card also has a piezo beeper that is used by the Multibus CPU card to signal the operator of error conditions and to provide audible feedback when a membrane button is pushed 2 5 5 2 INSTRUMENT DESCRIPTION CONTROL AND DISPLAY SUBSYSTEMS Cytometer Control Subsystem Figure 2 5 3 shows the Cytometer control paths of the Elite The Cytometer Control Subsystem consists of the Data Taker Interface card e Gated Amp Control card e Sensor I O card e Motor Control card DAC card Description Most of the Cytometers circuit cards have multiple functions and settings that must be configured by the Multibus CPU card The CPU places an address and a data word on the multibus back
339. for more information The necessary hardware to store the list gate window and to compare the appropriate datapath parameters resides on this card Note If it is possible to obtain acquisition on the Cytometer screen and not on the Workstation monitor then the problem is clearly isolated to one of the Lister cards or to the parallel cable or Lister A B in computer PN 4237003C PN 42370036 INSTRUMENT DESCRIPTION SIGNAL SELECTION AND DIGITIZATION Figure 2 13 9 Block Diagram Lister Cards Cpu uses data from LISTER OUT CARD this path to generate the four small All parameters for the histograms current event are stored here for the CPU to read Input buffer receives data from ADC lt FIFO REGISTER OUTPUT BUFFER and PSH control card from datapath bus TIME CIRCUITS LIST GATE Compare stored values to _ u SATISFIED current event parameter value STORE GATE VALUES LIST GATE CIRCUITS CONTROL CIRCUITS PARALLEL CABLE BETWEEN CYTOMETER AND WORKSTATION LISTER A B CARD BUFFER AND TEMPORARY INTERFACE STORAGE BUFFER INPUT BUFFER TO WORKSTATION 4K BYTES 7003064A Regarding Figure 2 13 9 Time circuits start counting elapsed time when acquisition starts The time for the current event can be appended to the data stream as the last parameter
340. formation Update form and will be released by service memo PN 4237003C iii REVISION STATUS iv PN 4237003 PN 42370036 LEGAL NOTICES REVISION STATUS iii 1 INTRODUCTION 1 1 1 1 MANUAL DESCRIPTION 1 1 1 Scope 1 1 1 Intended Audience 1 1 1 Organization 1 1 2 Format 1 1 2 Numbering Format 1 1 2 Special Headings 1 1 3 Warning 1 1 3 Caution 1 1 3 Important 1 1 3 Attention 1 1 3 Note 1 1 3 Conventions 1 1 3 SAFETY PRECAUTIONS 1 2 1 Electronic 1 2 1 Biological 1 2 1 Laser 1 2 2 Laser Beam Hazards 1 2 2 Radiation Hazards 1 2 2 Notify the Customer About Barrier Protection 1 2 9 2 INSTRUMENT DESCRIPTION 2 1 1 Sch 2 2 INSTRUMENT OVERVIEW 2 1 1 Function 2 1 1 Description 2 1 1 Main Components 2 1 1 Subsystems 2 1 1 Physical Specifications 2 1 2 Power Requirements 2 1 2 Reagents and QC Materials 2 1 2 Software 2 1 2 Fluorescence and Light Scatter Measurement Overview 2 1 2 Sensor Area 2 1 2 Sample Introduction System 2 1 3 AUTOCLONE SORTING OPTION 2 2 1 General 2 2 1 Sorting Tray Wells 2 2 1 Preventing Electrostatic Build Up 2 2 1 vi 2 3 ot 2 3 Mechanical Assembly Description 2 2 2 Motors 2 2 2 Tray Rotation Arm 2 2 2 Optical Sensors 2 2 2 Autoclone Sorting Option Card 2 2 3 Autoclone Sorting Option Interconnect Card 2 2 3 Position Detection Cards 2 2 3 Software 2 2 4 Calibration 2 2 4 Autoclone Sorting Option M
341. ft slot and adjust the larger 15 Vdc Analog Power Supply using a DVM as follows Black DVM Lead Red DVM Lead DVM Result Black test point Orange test point 15 0 V 0 05 Black test point Green test point 15 0 V 0 05 Vdc 15V 3 4 A supply Adjust the 15 Vdc 3 A power supply connected to Bertans with DVM connected to the HV backplane at P13 on top Bertrans backplane Put your black DVM lead on the black white wire Probe the green wire with the red DVM lead and adjust for 15 1 V 0 05 Vdc Move the red DVM lead to the orange wire Adjust for 15 1 V 0 05 Vdc 90 V power supply Adjust the power supply using the DVM as follows Black DVM Lead Red DVM Lead DVM Result OUT Upper Supply OUT Upper Supply 90 Vdc 2 Vdc OUT Lower Supply OUT Lower Supply 90 Vdc 2 Vdc Deflection power supply Not adjustable Test at test points J2 black ground J3 blue 2 3 to 3 0 Vdc J4 red 42 3 to 3 0 Vdc A 3 2 PN 4237003C FILE PATHS B 1 LOCATION OF SOFTWARE FILES Figure B 1 1 shows the location of software files Figure B 1 1 File Paths C ROOT DIR cla 2L d s r CONFIG SYS CTRL386 SYS RAMDRIVE SYS EMSTUB SYS T T T T MOUSE SYS AUTOEXEC BAT CGI CFG CONTROL HEX CONFIG SYS GSSCGI SYS CHAR4S FLT ADLISTER COM DRIVERS EXE CHAR9
342. g Solenoid operated Applies vacuum to either the Autoclone Sorting Option Option Drain double acting pinch valve waste collector or to the sample station drain VL14 Autoclone Sorting Solenoid Applies pressure to the Autoclone Sorting Option air Option Waste cylinder which pushes the waste collector forward VL15 Water Drain Solenoid operated Allows liquid in the air water separator to drain into the single acting pinch valve waste bottle when deactivated 2 9 2 Unless otherwise noted the valve performs when activated PN 4237003C INSTRUMENT DESCRIPTION 2 PNEUMATICS SUBSYSTEM Figure 2 9 1 illustrates valve operation Figure 2 9 1 Valve Operation PRESSURE IN Vent open Actuator is relaxed Coil m OOO M de energized n Pressure in Actuator H 1 is vented to atmosphere through the valve J lt passes through valve is blocked to Actuator Actuator is activated H Coil Energized Ee 24 V applied H Pressure moves piston inside the actuator applying force to the pinch valve INTERNAL OPERATION OF SOLENOID VALVE AND ACTUATOR LI SOLENOID VALVE C SYSTEM PRESSURE 2 PASSAGE Ex ACTUATOR PIN
343. g HeCd laser 8 2 33 8 1017262 Cover plate bottom HeCd laser 8 2 33 5 1017375 Plate rear mounting HeCd laser 8 2 33 4 6 1017501 Mount pinch valve analyzer 8 2 18 11 1017916 Base 8 2 17 9 1018879 Plunger test tube 8 2 18 3 1019539 Bracket interlock Elite Analyzer 8 2 20 1 8 1 12 42370036 MASTER PARTS LIST IN NUMERICAL ORDER CATEGORY Table 8 1 10 Hardware Continued PARTS LISTS Part Number Description Figure Item 1019658 Bracket air duct HeNe Argon Laser 8 2 35 2 1020271 Brackets mounting Argon laser 8 2 34 6 7 1020382 Bracket 8 2 3 Waste 12 Sheath 16 1020901 Holder pinch valve analyzer 8 2 18 21 1020903 Bracket tube holder Elite Analyzer 8 2 20 15 1022190 Screw 0 25 0 28 x0 94 HeCd Laser 74 8 2 33 2 2306051 Bracket mounting transducer 8 2 6 9 2508022 Coupling shaft 0 188 and 0 250 i d ends Autoclone Sorting Option 8 2 23 5 2515062 Spring analyzer 0 344 o d x 0 62 x 0 25 8 2 18 4 2523062 O ring sea 0 187 i d x 0 0501 8 2 18 18 2523549 Spring 0 250 o d HeNe Argon laser 8 2 35 3 2523656 Washer 0 25 bolt 0 63 o d HeCd Laser 74 8 2 33 3 2751587 Nut hex 25 20 UNC Argon laser 2851598 Nut hex 37 16 Argon laser 8 2 34 2 2851762 Screws machine metric M6x25 mm Argon laser 8 2 34 3 9 2806201 Screw 6 32 x 0 38 analyzer sample 8 2 17 8 2814004 Screw 25 20 x 0 50 HeNe Argon laser 8 2 35 14 2821010 Nut self locking analyzer 8 2 18 19 2822033 Nu
344. g a sample line or fiber optic cable 8 Reinstall the inner panel Figure 3 2 2 Dual CRT Installation AA ee 7003152A PN 4237003C 3 2 3 INSTALLATION PROCEDURES INITIAL SETUP Instrument Installation 1 Place the computer pedestal in its proper location 2 Install the table top as follows a Insert the table top into the brackets on the front of the Cytometer b Rest the free end of the table top on the computer pedestal C Secure the table top to the Cytometer with the screws provided 3 Remove the front cover from the computer pedestal and slide the computer into the pedestal from the front Verify that the computer power switch is at the top of the pedestal and is in the ON position Remove the keys from the back of the computer and put them in the filter box Place the color monitor mouse and keyboard in the correct locations on the table top Remove the rinse bottle holder Figure 3 2 3 a Open the pneumatics compartment door b Loosen the screws that hold the rinse bottle holder in place c Remove the rinse bottle holder Figure 3 2 3 Rinse Bottle Holder Location Remove all pinch valve clips before operation 7003001A 3 2 4 PN 4237003C INSTALLATION PROCEDURES 3 INITIAL SETUP 8 Position the Compressor module to the floor a Lift up on the Compressor module handle Remove the four scr
345. g adjustments must be made on the component side of the Quad 20 40 60 Microsecond Delay card slot10 Adjust R97 on the Quad 20 40 60 Microsecond Delay card slot 10 as above Change the delay to 40 microseconds and adjust R101 slot 10 as above Change the delay to 60 microseconds and adjust R105 as above AUX 2 Adjustment See Table 3 9 3 and Figure 3 9 4 for additional information on adjustments Change the delay to 7 microseconds At the Cytometers Scope screen select PARM B for the upper trace At the Cytometers Gated Amp Assign screen select AUX2 UPBM noting the amplitude as before Adjust R116 on the 7 Microsecond Delay card in slot 9 for the an amplitude of three divisions At the Cytometers Gated Amp Control screen select 20 microseconds delay Adjust R98 on the Quad 20 40 60 Microsecond Delay card slot 10 for an amplitude of three divisions Change the delay to 40 microseconds and adjust R102 for an amplitude of three divisions PN 4237003C PN 42370036 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE 8 Change the delay to 60 microseconds and adjust R106 for an amplitude of three divisions 9 Atthe Workstation put the protocol in RECEIVE CYTOSETTINGS AND SORT SETTINGS 10 Press to receive the high voltages and gains ll Press and change to the Protocol screen 12 Change to SEND CYTOSETTINGS AND SORT SETTINGS and save the protocol as GATED 13 Turn off the unit 14 Remove the Quad 20 40 6
346. g an enclosed bimorph Breakoff point is Last attached drop Leak in pneumatics system Find leak and repair unstable do REM UP Qld sample tubing Replace sample tubing with new Crack or slight clog in flow Inspect flow cell and replace if necessary cell Three minutes after the last attached Bimorph is unstable bien bimorph using an enclosed drop looks good it HIE BN varies again Breakoff is stable for Room temperature is too Check the daily temperature variation and 1 to 2 hours then high ensure the operating temperature is 18 29 C never returns to the 64 84 F Adjust air conditioning as needed same place until the next morning Note A room temperature change of gt 1 C in 1 hour can cause a change in droplet formation Sample run When RUN is Air in the sample line 1 Remove the sample and press CLEAR affects breakoff point pressed and sample push is over the breakoff point moves down one drop Disturbance in the sample flow as it enters the sheath several times to get the last droplet back to the reference cursor 2 Ensure the side streams are stable and there is no wisping 3 Without a sample on press RUN to backflush the sample line and to clear out the remaining air bubbles While still in RUN put the sample on The sample is automatically run the push occurs for the specified time frame and the last attached drop should not move PN 4237003C
347. g performance Beckman Coulter recommends the room temperature not fluctuate more than 1 C 3 5 F in an hour during sorting Space and Accessibility The Elite standard configuration consists of e One 15 mW air cooled Argon laser and One 10 mW HeNe laser The configuration for an Elite with optional laser s is the same as the standard configuration plus e HeCd laser and or e Water cooled laser See Table 3 1 4 for the unit size and necessary operating clearances for each configuration PN 4237003 3 1 3 INSTALLATION PROCEDURES PREINSTALLATION CHECK Table 3 1 4 Required Operating Clearance Specifications Instrument Configuration Elite Standard Configuration Elite with Optional Laser s t Width Additional clearance on right Additional clearance on left Total clearance needed 142 cm 56 in 46 cm 18 in 5 cm 2 in 192 cm 76 in 173 cm 68 in 46 cm 18 in 15 cm 6 in 234 cm 92 in Length Additional clearance in rear Total clearance needed 170 cm 67 in 46 cm 18 in 216 cm 85 in 170 cm 67 in 46 cm 18 in 216 cm 85 in Height Additional clearance on top Total clearance needed 130 cm 51 in 46 cm 18 in 176 cm 69 in 130 cm 51 in 46 cm 18 in 176 cm 69 in Consists of one 15mW air cooled Argon laser and one 10 MW HeNe Laser 1Same as standard configuration plus an additional HeCd laser and or w
348. g the adjustable HeNe laser dichroic on the optical rail Look at the lower trace on the Digiscope Screen and e Adjust for 20 microseconds for a 100 p quartz tip e Adjust for 30 microseconds for a 76 p quartz tip e Adjust for 3 5 microseconds for a 76 p jet in air tip Note The UV signal is on the far left on the Digiscope screen Test this by increasing decreasing the UV laser power and observing the far left peak Close the HeNe laser output shutter Install the adjustable beam translator between the Argon laser and its adjustable dichroic leaving room between the translator and the optical rail to mount a single laser target Verify the following a The translator knobs face away from laser b The translator window is perpendicular to the optical bench Adjust the translator height in the post for the laser to penetrate as close to the center of the translator window as possible Refer to the multi hole target and set the alignment mode for lowest target hole Move pin on bottom of target to line up with desired target hole and lock the pin down with the setscrew Install the lowest target in the space you left between the beam translator and the Argon dichroic ensuring the lowered target is square with the table top in all respects Rotate the translator window so the Argon beam is aligned with the lowest laser target hole Remove the lowered Argon laser target Adjust between the peak of the Argon laser generated s
349. ge Elite See Table 8 2 32 7003098A Table 8 2 32 Elite Flow Cell Stage Detail of Parts See Figure 8 2 32 Item Part Number Description 1 3814271 Ultra compact positioner 2 1010932 Upper nut insertion rod gland nut 3 6856762 Flow cell body high optical 4 6232208 Hose barb fitting elbow 0 093 i d nylon white 5 6602642 Biosense flow cell 6 6858368 Bimorph flow cell bracket 8 2 32 PN 4237003C PARTS LISTS 8 ILLUSTRATED PARTS Figure 8 2 33 Model 74 HeCd Laser Mounting Hardware See Table 8 2 33 FRONT OF INSTRUMENT y 00 7003006A Table 8 2 33 Model 74 HeCd Laser Mounting Hardware Detail of Parts See Figure 8 2 33 Item Part Number Description 1 6705218 HeCd 74 Laser and Head tested 2 1022190 Screw 0 25 x 0 28 x 0 94 3 2523656 Washer 0 25 4 1017375 HeCd mounting back plate 5 1017262 Plate bottom cover 6 1016827 Plate support upright rear 7 1017261 HeCd mounting front plate PN 4237003C 8 2 33 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 34 Argon Laser Mounting Hardware See Table 8 2 34 7003010A Table 8 2 34 Argon Laser Mounting Hardware Detail of Parts See Figure 8 2 34 Item Part Number Description 1 3814232 Argon lon system laser 2 2851598 Hex nut 37 16 3 2851762 Machine screws metric M6x25 mm 4 2826051 Washer split lock 5 6028218 Cable assembly with interlock 6 10202
350. h Back Porch length 31 25 microseconds Center level length number of droplets to sort 1 Front and Back porches 0 to 10096 of Center level Crystal Drive Test Procedure Wave form sinusoidal measuring 160 V peak to peak 5 V with little distortion Crystal Drive Adjustment Procedure Crystal drive 10096 Drop frequency 32 kHz Adjust R84 on the Sort Oscillator card for 8 5 V peak to peak Adjust R118 on the Sort Output card for 160 V 5 V peak to peak signal Sort Pulse Amplitude Test Drop frequency 32 kHz Sort Test ON Sort Left Enable ON Sort Right Enable ON Sort Right counter OFE Left Stop counters OFE Deflection 10096 Drops sorted 3 Signal baseline 0 Vdc Amplitude of center level from the baseline 80 V 2 5 V Negative going pulse symmetrical with positive going pulse Front and Back porches varied from 096 to 10096 Front and Back porches back to reference values Front porch at 8096 Rear porch at 2096 PN 4237003 A 3 1 QUICK REFERENCE INFORMATION OPTICAL SETTINGS Power Supplies Table A 3 1 Power Supply Adjustments Fiber Optics supply Turn to minimum intensity Switcher supply Adjust for 5 25 Vdc 0 01 Vdc between yellow and black wires 15 V 0 25 A PMT Adjust for 15 0 0 05 Vdc on orange wire 15 0 0 05 Vdc on green wire Both referenced supply to black wire 15 V 5 0 A supply Locate the Pulse Generator and Clock card in the Data Acquisition lower backplane far le
351. he MUX J23 end of coax to Mux and Scope J23 Connect the MUX J24 end of coax o Mux and Scope 24 Relabel and install the following 27 in cables as indicated in Table 3 11 1 Table 3 11 1 Card and Cable Connections From To 3 Sub 1 FL3 Int Out J8 Mux and Scope Int J16 3 Sub 1 FL3 Log Out J7 Mux and Scope Int J17 3 Sub 1 FL3 Pk Out J6 Mux and Scope J18 Dual FL FL2 Out J7 Mux and Scope J19 Dual FL 2Log Out J8 Mux and Scope J20 Dual FL FL2 Pk Out J6 Mux and Scope J21 Note Text contained in parentheses indicates silkscreen information on circuit cards NOT cable labeling PN 4237003C 3 11 3 INSTALLATION PROCEDURES 5TH PMT KIT WITH SWITCHABLE AMPS INSTALLATION PROCEDURE 3 11 4 Software Installation Verify that system software is at 4 01 or above If not obtain and install current software See Heading 4 9 SOFTWARE INSTALLATION for instructions System Test 1 Remove the 488 DL filter and replace with a full mirror 2 Remove the following filters for this test so that only 45 filter elements are present in PMT block 488 LB 525 BP 575 BP 675 BP 488 BP 3 Reset those cards where coax cables were removed and installed from 4 Turn instrument power ON 5 Ensure column for 5th PMT HV INT Gain PK Gain is present and operational 6 Atthe Control Align screen select a Align LED b Pulse Mode c 100 Drive d 1kHZ 7 At the Scope Screen a S
352. he knob on the deflection body holder to center the ground plate opening around the stream Ifthe stream is still too close to one side the other Z axis of the flow cell stage is not adjusted correctly Ensure that this is set correctly through proper alignment before continuing Any movement of the Z axis can affect the rest of the optical alignment e After a new flow cell flow body or bimorph stage is replaced you should only need to adjust the knob once 6 SettheSort settings Refer to Drive Frequency Phase Adjustment and Delay Calculation in this section for details 7T Sorta mixture of different brightness beads 50 000 right and left Refer to the settings obtained in step 6 Refer to Figure 3 6 1 for placement of tubes for sorting e non ESP systems sort at 1 500 events sec data rate with Abort ON For ESP systems sort at 5 000 event sec with Abort ON Complete Abort Figure 3 6 1 Tube Placement for Sorting ma SS WASTE STREAM COLLECTOR BEAK SORT TUBE HOLDERS COMPARTMENT FLOOR 7003124A 36 2 4237003 INSTALLATION PROCEDURES 3 SYSTEM TESTING 8 Reanalyze the sorted beads to determine purity e gt 95 for non ESP systems gt 99 for ESP systems Drive Frequency l Setthe Drive Frequency to 32 kHz 2 Set the Crystal Drive to 7596 3 View the droplets above the ground plate and scan through the frequency settings using the tenth decimal adjustmen
353. he sheath pressure sensor is replaced Figure 4 4 1 shows the location of the power supplies Tools Supplies Needed Digital pressure gauge accurate to 0 01 psi DVM Figure 4 4 1 Power Supplies PMT HV SUPPLY FIBEROPTICS 36 V SUPPLY SUPPLY circuit SUPPLY FAN 107 CFM 115 VAC 3000 DEFLECTION SUPPLY 15 V 0 25 A SUPPLY 15 V 5 0 A SUPPLY 90 V SUPPLY 15 V 3 4 A SUPPLY 7003068A REAR VIEW Regarding Figure 4 4 1 The PMT HV power supply is controlled by the software e The switcher supply provides 5 V to all logic circuits 12 V for multibus and 24 V for solenoids and motors The fiber optics supply should be turned to minimum intensity The 90V supply goes to the Sort Output card 4237003 4 4 1 4 SERVICE AND REPAIR PROCEDURES POWER SUPPLIES MEASUREMENT AND ADJUSTMENT The Deflection supply is not adjustable Test at the following test points Procedure J2 Black Ground J3 Blue 2 3 to 3 0 Vdc J4 Red 2 3 to 3 0 Vdc Do the measurement and adjustment procedures in Table 4 4 1 Table 4 4 1 Power Supply Measurement and Adjustments Power Supply Perform Fiber optics supply Turn to minimum intensity Switcher supply Adjust to 5 25 0 01 Vdc between yellow and black wires 15 V 0 25 A PMT supply Adjust for 15 0 0 05
354. he way down stream and there is 19 20 sufficient space above the ground plate at least one droplet a Turn on the Sort Left b Turnon Sort Right Activate the sort test d Turnon the High Voltage Ensure that 3 drops sorted is selected and view the side streams If the light is not sufficient to see the streams adequately adjust the light so the side streams can be monitored at a glance Using the phase adjustment of the droplet delay button scroll through the settings and ensure that at least seven or eight out of the 10 settings produce tight clean singular side streams a If the streams are acceptable go to step 21 b Ifthe streams are not acceptable 1 Check the filament between the last attached and the next one up If the filament is not acceptable adjust crystal drive PN 4237003C 21 22 23 24 PN 4237003C SERVICE AND REPAIR PROCEDURES SORT SENSE FLOW CELL ADJUSTMENT AND VERIFICATION 2 Check the flow cell and or flow cell body Turn off the HV to the sort plates and press CLEAR two times After the droplets return to their position check the side streams again If they do not return to the same position there may have been a clog or air bubble in the flow cell 3 Verify the crystal frequency setting to ensure that it was not set incorrectly due to a fluidics problem 4 Correct the fluidics problem if applicable Stability is very important If the last droplet still doe
355. hed to a metal part of the chassis connected to an earth ground Biological WARNING Risk of personal injury or contamination If you do not properly shield yourself before servicing the instrument with the door open you may be injured or contaminated To prevent possible injury or biological contamination you must wear gloves a lab coat and eye protection when servicing the instrument with the doors open and or when working with pathogenic materials Use extreme care when working with pathogenic materials Means must be available to decontaminate the instrument to ventilate air and to dispose of waste liquid Refer to the following publications for further guidance on decontamination Biohazards Safety Guide 1974 National Institute of Health e Classifications of Etiological Agents on the Basis of Hazards 3d ed June 1974 Center for Disease Control U S Public Health Service PN 4237003 1 2 1 INTRODUCTION SAFETY PRECAUTIONS 1 2 2 Laser Laser Beam Hazards The laser is a unique light source that shows characteristics different from those of conventional light sources The safe use of the lasers depends upon familiarity with the instrument and with the properties of coherent intense beams of light Because the Elite contains at least one laser the customer should keep the instrument isolated from nonlaser instruments The customer should also keep a copy of ANSI Standard 136 1 SAFE USE OF LASERS near the
356. hen varies the voltage applied to the sheath pressure regulator to obtain the requested value This system differs from the Sample Pressure Regulation circuit in that the sheath pressure sensor actually measures the pressure present in the liquid line between the sheath tank and the flow cell The reason for this is that the system must adjust the air pressure in the sheath tank to compensate for the change in liquid head pressure as the sheath level drops during operation For this reason you should make all adjustments when the sheath tank is full The sheath pressure sensor requires a 10 V power supply to operate The R54 sets this requirement and you can measure it at TP3 Instrumentation type Op Amp is interfaced to the sensor This circuit provides both gain and offset adjustment for the sensor input You can measure the output at TP4 which is the voltage provided for the Sensor Interface card for digitization and display on the screen as the read sheath pressure The CPU writes the desired sheath pressure to the Pneumatic Interface card The DAC converts the information to an analog voltage The value of the DAC reference voltage as measured at TP10 and adjusted by R81 determines the conversion scaling The op amps that drive the sheath pressure regulator compare the DAC output with the amplified sensor feedback value to apply the appropriate voltage to the regulator to achieve the requested PN 4237003C PN 42370036 PNEUMATICS S
357. herent 6913212 HeNe laser Melles Griot Green 6913229 HeCd 74 Omnichrome laser 6913250 Sort enhancement switch amp after SN U07122 6913251 Snout upgrade 6913253 Sort enhancement pre switch amp before SN U07122 6914966 Software driver HP 1600C for International use only 6914967 HP 1600C Color Printer ink cartridges 6914968 HP 1600C Color Printer 6914969 Printer color Elite 6915005 90 MHz Pentium full workstation upgrade 6915006 90 MHz Pentium upgrade Elite 6915082 Elite 256 Sort Upgrade Table 8 1 12 Lamps Part Number Description Figure Item 3908021 Lamp indicator 24V amber 3908022 Lamp indicator 24V red 3908023 Lamp indicator 24V green 3908025 Lamp halogen 21 V 150 W 8 2 13 9 Table 8 1 13 Lasers Part Number Description Figure Item 3203037 Foam strip panel laser interlock assembly 3814232 Argon lon laser 8 2 34 1 3814233 Table top optical Argon laser 8 2 34 8 3814244 Power supply HeCd laser 8 2 38 1 3814286 Mirror 325 8 2 36 4 6027225 Line cord Omnichrome HeCd laser 6028256 Cable interlock Omnichrome HeCd laser 8 2 38 4 6705218 Laser HeCd 74 8 2 33 1 6856681 Upright mounting Argon laser 8 2 35 12 8 1 16 PN 4237003C MASTER PARTS LIST IN NUMERICAL ORDER CATEGORY Table 8 1 13 Lasers Continued PARTS LISTS Part Number Description Figure Item 6856859 Table top laser mount 8 2 4 1 Laser Alignment Targets With 3 Target Holes 6857274 Large 8 2 36 9 6857236 Small 8
358. hin the region and a 0 value if it is out of the region The Workstation then transfers the single bit representation bitmap to the Cytometer The Cytometer sends the pattern via the DT Interface card to one of the bit map memories During the actual sort the system tests each cell Essentially the card uses the channel numbers for the two appropriate parameters as coordinates to access the stored bit map and to read the stored bit If the system reads the bit as a 1 then the event has fallen within the amorphous region If the card does not read the bit as a 1 then the event has not fallen within the amorphous region A considerable amount of logic circuitry is needed latch in the correct parameter channel numbers as they appear on the dataPath bus To use these values as memory addresses into the proper bitmap memory To latch out the appropriate results The card then compares the results to the stored sort criteria in order to reach a second partial decision to sort based on bitmaps The decision to sort based on bitmap comparisons and sort logic are combined with the linear region results to reach a final sort decision which is does the cell meet all gate and region conditions set for a right or left sort The card sends this decision to the Sort Delay card for synchronization to system droplet generation PN 4237003C INSTRUMENT DESCRIPTION SORT SUBSYSTEM Figure 2 8 4 Block Diagram Bitmap and Sort Deci
359. hort pass function 2 3 12 optical plate See plate optical Optics PN 4237003C INDEX alignment components 4 12 2 collection See collection optics troubleshooting procedures 7 1 5 optics operational test See VIP P pan feature See camera parallel plate function of 2 7 7 part numbers list of 8 1 1 listed by category 8 1 1 parts list of 8 1 1 parts list by category See part numbers illustrated 8 2 1 pathways of display sources 2 5 4 PC Model 486 ADVANCED CMOS SETUP 4 8 1 Pentium 133 MHz BIOS setup 3 21 3 Pentium 133 MHz technical specifications 3 21 1 Pentium 133 MHz upgrade 3 21 1 PERIPHERAL MANAGEMENT SETUP 4 8 2 setup procedure 4 8 1 STANDARD CMOS SETUP 4 8 1 Peak ADC and PSH Control card jumpers 4 10 3 A 2 3 Peak ADC card ASD settings illustration of 2 13 9 function 2 13 9 timing illustration of 2 13 10 Peak ADC PSH Control card block diagram 2 13 7 jumpers 4 10 3 A 2 3 Peak Scatter Sensor card connections 3 9 2 Peak Scatter Mux SW R card block diagram 2 10 4 function 2 10 3 jumpers 3 9 4 4 10 2 4 10 3 A 2 2 A 2 3 signal flow summary of 2 6 9 phase adjustment procedure 3 6 3 phase settings adjusting for sorting 4 2 5 requirements 3 6 3 verifying a shift in the crystal drive 4 2 3 Piezo bimorph beeper function of 2 5 5 INDEX 11 INDEX flow cell alignment 3 16 2 installation procedure 3 16 1 when to install 3 16 1 Piezoelectric transducer See Piez
360. hree lasers 4 1 6 time space separated laser 3 12 5 4 1 7 analog calculator circuit defined 2 13 5 analog data ADC card received by 2 13 8 ASD calculations 2 13 10 change requirements 2 13 8 defined 2 13 8 function 2 13 8 settings illustration of 2 13 10 Autoclone calibration requirements 2 2 4 Interconnect card See Autoclone Interconnect card valve drain VL13 function of 2 9 2 Autoclone Sorting Option calibration requirements 2 2 4 card See Autoclone Sorting Option card CC marker defined 2 2 3 CW marker defined 2 2 3 electrostatic buildup in sort wells preventing 2 2 1 Hx marker defined 2 2 3 Hy marker defined 2 2 3 illustration mechanism 2 2 1 inspection electrical 3 10 2 inspection mechanical 3 10 2 Interconnect card See Autoclone Sorting Option Interconnect card Lx marker defined 2 2 3 Ly marker defined 2 2 3 markers screen definitions of 2 2 3 mechanism adjustment procedure 2 2 4 mechanism function 2 2 1 motors function of 2 2 2 optical sensors status of 2 2 2 path through tray wells 2 2 1 Position Detection cards See Position Detection 2 INDEX cards sensor optical function of 2 2 3 sensors defective result of 2 2 3 sensors function of 2 2 2 sensors optical function of 2 2 2 software description 2 2 4 spade lug connections 3 10 5 speed adjustments 3 10 7 stream adjustments 3 10 9 switch limit function of 2 2 3 switch mechanical function of 2 2
361. hysical Specifications 2 1 2 Power Requirements 2 1 2 Reagents and QC Materials 2 1 2 Software 2 1 2 Fluorescence and Light Scatter Measurement Overview 2 1 2 Sensor Area 2 1 2 Sample Introduction System 2 1 3 22 AUTOCLONE SORTING OPTION 2 2 1 General 2 2 1 Sorting Tray Wells 2 2 1 Preventing Electrostatic Build Up 2 2 1 Mechanical Assembly Description 2 2 2 Motors 2 2 2 Tray Rotation Arm 2 2 2 Optical Sensors 2 2 2 Autoclone Sorting Option Card 2 2 3 Autoclone Sorting Option Interconnect Card 2 2 3 Position Detection Cards 2 2 3 Software 2 2 4 Calibration 2 2 4 Autoclone Sorting Option Mechanism Adjustment 2 2 4 23 OPTICAL COMPONENTS 2 3 1 Lasers 2 3 1 Beam Shapers 2 3 5 Beam Steering Optics 2 3 7 Beam Translators 2 3 8 Beam Expander Reducer 2 3 8 Side Scatter and Fluorescence Collection Optics 2 3 9 Sort Sense Flow Cell Tips 2 3 9 Jet in Air Flow Cell Tips 2 3 10 Fast Quartz Flow Cell 2 3 11 Optical Filters 2 3 11 Long Pass LP Filters 2 3 12 Short Pass SP Filters 2 3 12 Blocking BK Filters 2 3 12 Band Pass BP Filters 2 3 12 Dichroic Long Pass DL Filters 2 3 12 Dichroic Short Pass DS Filters 2 3 13 Neutral Density ND Filters 2 3 13 Forward Scatter Detector 2 3 13 Forward Scatter Obscuration Templates 2 3 13 PN 4237003 2 i 24 2 3 2 6 da ELECTRONICS SUBSYSTEM 2 4 1 Description 2 4 1 CONTROL AND DISPLAY SUBSYSTEMS 2 5 1 Mu
362. i ILLUSTRATIONS 1 2 1 Laser Warning Labels Coherent Innova 300 Water Cooled Argon Laser 1 2 3 1 2 2 Laser Warning Labels Innova 90 Water Cooled Argon Laser 1 2 4 1 2 3 Laser Warning Labels Helium Cadmium Laser 1 2 4 1 2 4 Laser Warning Labels Omnichrome Helium Cadmium 74 Laser 1 2 5 1 2 5 Laser Warning Labels Coherent Innova Enterprise Laser 1 2 5 1 2 6 Laser Warning Labels Coherent Innova 305 Laser 1 2 6 1 2 7 Laser Warning Labels Coherent Spectrum Laser 1 2 6 1 2 8 Laser Warning Labels Uniphase Red HeNe Laser 1 2 7 1 2 9 Laser Warning Labels Melles Griot Green HeNe Laser 1 2 7 1 2 10 Laser Warning Labels Sensing Area 1 2 8 2 1 1 Elite Components 2 1 1 2 1 2 Sensor Area 2 1 3 2 1 3 Sample Stage 2 1 3 2 2 1 Autoclone Sorting Option Mechanism 2 2 1 2 3 1 Optical Components Top View with Cover Removed 2 3 1 2 3 2 Beam Shaper Options 2 3 6 2 3 3 Alignment Mirror 2 3 8 2 3 4 Beam Expander Reducer 2 3 9 2 3 5 Sort Sense Flow Cell Original 2 3 10 2 3 6 100 Sort Sense Flow Cell Cut away View 2 3 10 2 3 7 76 pm High Velocity Quartz Flow Cell 2 3 11 2 3 8 Optical Filters Location 2 3 12 2 3 9 Forward Scatter Detector 2 3 13 2 3 10 Forward Scatter Obscuration Templates 2 3 14 2 4 1 Block Diagram System Electronics 2 4 3 2 5 1 Laser Control Subsystem 2 5 3 2 5 2 Block Diagram Video Subsystem 2 5 4 2 5 3 Control Pathways 2 5 7 2 6 1 Subtraction Circuit Example 2 6
363. ial 1 2 1 defective fittings See fittings deflection power supply adjustments and measurements 4 4 2 A 3 2 delay calculations for sorting 4 2 7 procedure 3 6 5 delay function how to set for sorting 4 2 3 settings 4 2 3 delays Gated Amp card set by 2 12 2 dichroic long pass DL filter function 2 3 12 dichroic short pass DS filter function 2 3 13 Digiscope card function 2 5 5 jumpers 4 10 2 A 2 2 digital values circuit cards that read 2 13 1 digital to analog converter See DAC diode capacitors charging 2 13 4 discriminator circuit See circuit discriminator discriminator extension See ASD DISCSAT defined 2 13 4 function of 2 13 2 2 13 7 DL dichroic long pass filter function 2 3 12 drip chamber biofilter function of 2 9 1 drip chamber drain VL12 function of 2 9 2 Drip Chamber Error 2 9 1 drive frequency procedure 3 6 3 settings 3 6 3 drop delay setting calculating 4 2 7 quarter drop sort defined 2 8 8 DS dichroic short pass filter function 2 3 13 Dual CRT Control card function 2 5 5 4237003C jumpers 4 10 2 dual CRT See CRT dual Dual FL Amp card block diagram 2 11 3 FL inputs 2 11 3 function 2 11 3 Dual FL Amp SW R card jumpers 4 10 2 4 10 3 A 2 2 A 2 3 Dual FL SW R card block diagram 2 10 2 function 2 10 1 jumpers 3 9 3 Dual Laser Control card function 2 5 2 jumpers 4 10 2 A 2 2 dyes emissions 2 6 6 requirements 2 6 6 E electronics opera
364. ible display e Receives stretched pulses from the outputs of the PSH cards The card can select two of these pulses for possible scattergram display Is controlled by the Multibus CPU card For Gated Amp systems two additional inputs are used One allows display of the Gate Window or Gated Amp Trigger pulse for adjustment and alignment purposes The second connection provides the Gated Amp card access to the DISCSAT bus to implement the Gate Amp Discriminator Override function 2 13 2 PN 4237003C INSTRUMENT DESCRIPTION 2 SIGNAL SELECTION AND DIGITIZATION Figure 2 13 1 Block Diagram Mux and Scope Card MULTIPLEXER Chips Select output from All input lines go to each the 16 possible inputs Inputs Multiplexer chip gt gt o To PSH Cards 0 EE J31 Gate Amp Pulse from Gated Amp Control Card z Upper Trace pulses to J30 goes connection under table for use with external scope Lower Trace pulses to J29 goes to BNC connection under table for use with external scope Gate Pulse is put on backplane through connector P1 50 This is the DISCSAT bus Ribbon Cable to D Digiscope o R J27 4 J25 EX1 Input o Upper Trace X Ex1 W J27 2 J26 EX2 Input 9 Lower Trace Y Ex2 Select Y Choose pulse or X Y display Select X Stretched Pulses from PSH cards through Se
365. ick connect tubing connectors to the appropriate connector Be sure to match the colors b Plug the level sense connector into the matching socket PN 4237003C 3 4 3 INSTALLATION PROCEDURES CONNECTING ASSEMBLIES Figure 3 4 3 Sheath and Waste Bottles QUICK CONNECT TUBING CONNECTOR LEVEL SENSE CONNECTOR WASTES 7003003A Options Connections Laser Option See Heading 3 7 OPTIONAL LASER INSTALLATION for instructions Optical Drive Option The Workstation can be equipped with a number of mass storage drives For installation information refer to the information supplied with the drives Printer Option Connect the Printer to the PRINTER port on back of the Workstation s computer For additional information refer to the documentation supplied with the Printer 3 4 4 42370036 INSTALLATION PROCEDURES 3 TESTING ASSEMBLIES 3 5 TESTING ASSEMBLIES Cytometer Control Computer 1 Connect the two Cytometer power plugs to the ac outlets Turn ON the lower power switch on the Cytometer 2 3 Verify that the main control screen right Cytometer screen appears within 10 seconds 4 Verify that all cooling fans Figure 3 5 1 are operating With a reference point from the front of the instrument fan locations are Two on the right rear door One on the left rear door One above the Gate
366. id not make any other adjustments Figure 3 6 2 ESP Front and Back Porch Adjustment Knobs 15 16 NORMA WIDTH LIZATION 5 RECOMMEN RONT Porgy SETTINGS 8 ORCH 2 7003125A Check the side and center streams on the 1 2 and 3 Drops Sorted settings Note The streams should look good on each of these settings If not you may have to either clean or change the flow cell Adjust the sort pulse shape after the best possible Sort Test streams are achieved Note You can observe the sort pulses with your scope using TP2 on the Sort Output card Delay Calculation l Oy une de Ensure that the proper flow cell is selected on the cytometer Options screen The flow cell options include e JD um e 100 pm e Jet in air Calculate the droplet delay setting from the Camera screen of the Cytometer Refer to the Sorting section of the Operators Guide for instructions Go to the Sort screen Run the DNA Check beads to set a sort gate on a fluorescence signal Ensure that the Droplets Sorted is set to 1 Set the Sort Stop to 100 This makes it easy to see the correct drop quickly in the fluorescent microscope 3 6 5 INSTALLATION PROCEDURES SYSTEM TESTING 3 6 6 10 Run a sort seven times once for the actual Delay setting three at less than the actual setting and three at more than the actual setting on the Camera screen Sort onto the slide at each setting View the sort
367. if PMT 5 is installed Turn on the instrument and after the Elite Software to loads select the GATED AMP protocol At the Workstation s Parameter screen select FALS PARAMETER LOG Press At the Sort screen select Strobe and set Frequency to 2 kHz Be sure the deflection body is all the way up and the FALS filter is removed At the Sort Gated Amp Control screen a Select PMT1 PEAK for the trigger source b Setthe trigger discriminator to 100 c Adjust the trigger gain to see the lower scope trace three divisions tall Put mirror under PMTI and adjust HV to get 3 divisions PN 4237003C PN 42370036 10 11 12 13 14 15 16 17 18 19 20 21 22 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE At the Scope screen a Setlower scope trace to TRIG b Set upper scope trace to FALS LOG and 10 microseconds division At the Gated Amp Control screen a Raise the gain and voltage so PMT1 is displayed b Setthe clock for 0 5 microseconds Setthe window width to 5 5 microseconds d Setthe window delay to 5 5 microseconds and enable e Setthe signal delay to 7 microseconds Change the display or scope trace to display window At the Gated Amp Assignment screen Figure 3 9 6 a Select FALS LWBM b Block the FALS sensor totally so it is totally covered and dark Connect the scope to the upper trace UT on the connector panel BNC next to the ac under table top left of
368. if time is selected as a parameter Control Circuits determine if data leaves the Lister card Acquisition ON OFF The control circuits also select parameters to be tested for the list parameters to be tested for the list gates stores the gate region values and determines how many parameters to send for each event 2 13 13 2 INSTRUMENT DESCRIPTION SIGNAL SELECTION AND DIGITIZATION FIFO Register stores all the parameter values for the current event Values are only readout if the list gate is satisfied e Output Buffer sends the data to the Workstation Input Buffer receives the data from the Cytometer Lister A B Card This card resides in the Workstation computer The Lister A B card e Receives the acquired data from the Cytometer e Provides a buffer and interface to the Workstation computer 2 13 14 PN 4237003 CONTENTS 8 3 INSTALLATION PROCEDURES 3 1 1 PART A INSTRUMENT INSTALLATION 3 1 PREINSTALLATION CHECK 3 1 1 Education Center Training 3 1 1 Cartons Received 3 1 1 Power Requirements 3 1 1 Water 3 1 2 Drainage 3 1 3 Environment 3 1 3 Space and Accessibility 3 1 3 Optional Printer 3 1 4 Labels 3 1 4 Supplies 3 1 4 3 2 INITIAL SETUP 3 2 1 Unloading and Moving the Instrument 3 2 1 Dual CRT Installation 3 2 2 Instrument Installation 3 2 4 3 3 CYTOMETER CHECK 3 3 1 3 4 CONNECTING ASSEMBLIES 3 4 1 Electronic Cable Connections 3 4 1 Tubing Connections 3 4 3 Options
369. ify that the delay observed varies as the delay is varied on the touch screen Note Jitter of less than or equal to 1 drop is expected in the sort pulse timing This is normal since the sort pulses are synchronous with the crystal drive Sync Adjustment 1 Connect one scope channel to the crystal drive and the other to the sort pulses 2 Syncthe scope on the sort pulses 3 Verify that as in the sort test that the sort pulses are synchronous with the crystal drive Sort Function Verification IMPORTANT Risk of compromising purity and recovery if the sort verification procedures are not strictly followed To obtain a stable successful sort carefully follow each sort verification procedure 1 Ifinstrument has been shutdown a Prepare and start up instrument for daily operation as instructed in the Operators Guide b Check for proper alignment and flow c Perform any cleaning or adjustments at this point If saline is evident clean the deflection plates Plug in defection plates by placing the top of the strobe between the bottom of the beam shaping lens and the bottom of the lens holder 4 Center the stream within the sort plate opening by adjusting the knob on the deflection body holder to center the ground plate opening around the stream IMPORTANT Risk of misalignment Any movement of the Z axis of the flow cell stage can affect all other optical alignments This knob should only require adjustment once and that
370. ight Rear Door Inside View See Table 8 2 14 8 2 14 8 2 15 Cytometer Left Rear Door Inside View See Table 8 2 15 8 2 15 8 2 16 Cytometer Left Front Corner Inside View See Table 8 2 16 8 2 16 8 2 17 Analyzer Sample Drawer See Table 8 2 17 8 2 17 8 2 18 Analyzer Top Inside View See Table 8 2 18 8 2 18 8 2 19 Flow Cell Area Elite See Table 8 2 19 8 2 19 8 2 20 Flow Cell Area Elite Analyzer See Table 8 2 20 8 2 20 8 2 21 Deflection Body and Holder See Table 8 2 21 8 2 21 8 2 22 Photomultiplier Tubes PMTs See Table 8 2 22 8 2 22 8 2 23 Autoclone Sorting Option Mechanism See Table 8 2 23 8 2 23 8 2 24 Autoclone Sorting Option Beak Waste Catcher See Table 8 2 24 8 2 24 8 2 25 Autoclone Sorting Option Location and Connections See Table 8 2 25 8 2 25 8 2 26 Autoclone Sorting Option Accessories See Table 8 2 26 8 2 26 8 2 27 Transformer Drawer Assembly See Table 8 2 27 8 2 27 8 2 28 Computer Box Assembly See Table 8 2 28 8 2 28 8 2 29 Beam Splitter Dichroic See Table 8 2 29 8 2 29 8 2 30 Beam Shaper See Table 8 2 30 8 2 30 8 2 31 Flow Cell Stage Elite ESP See Table 8 2 31 8 2 31 xxi xxii 8 2 32 8 2 33 8 2 34 8 2 35 8 2 36 8 2 37 8 2 38 B 1 1 Flow Cell Stage Elite See Table 8 2 32 8 2 32 Model 74 HeCd Laser Mounting Hardware See Table 8 2 33 8 2 33 Argon Laser Mounting Hardware See Table 8 2 34 8 2 34 HeNe Laser Mounting Hardware See Table
371. ight by wavelength they block all wavelengths equally Use ND filters in front of the forward scatter detector to prevent detector saturation Forward Scatter Detector The forward scatter lens collects light that is scattered between 1 4 degrees and 19 degrees from the axis of the laser beam The forward scatter light is extremely bright A neutral density filter must be between the lens and the photovoltaic cells to reduce the intensity A bar on the FALS MASK in front of the detector blocks the laser beam See Figure 2 3 9 Figure 2 3 9 Forward Scatter Detector OBSCURATION BAR CAMERA PRISM 5904021A Forward Scatter Obscuration Templates Different applications may require different obscuration templates Figure 2 3 10 shows the available templates 2 3 13 2 INSTRUMENT DESCRIPTION OPTICAL COMPONENTS Figure 2 3 10 Forward Scatter Obscuration Templates FULL CIRCLE STANDARD CROSS QUARTZ 394 HIGH SENSITIVITY QUARTZ 394 R LOW SENSITIVITY JET IN AIR 394 QUARTZ 295 HIGH SENSITIVITY QUARTZ 295 R LOW SENSITIVITY JET IN AIR 295 NX NI RING 295 394 R QUARTZ 197 R HIGH SENSITIVITY QUARTZ 197 R LOW SENSITIVITY JET IN AIR 197 R e 7 197 295 QUARTZ 098 HIGH SENSITIVITY 60 QUARTZ 098 LOW SENSITIVITY JET IN 098 RING 098 197 QUARTZ HIGH SENSITIVITY QUARTZ LOW SENSITIVITY JET IN AIR 15mm
372. ignal and the peak of the UV laser generated signal by using the adjustable Argon dichroic on the optical rail Look at both traces on the Digiscope screen and e Adjust for 20 microseconds for a 100 p tip e Adjust for 30 microseconds for a 76 tip e Adjust for 3 5 microseconds for a 76 p jet in air tip Note The peak of the UV laser generated signal is the far middle one of the Digiscope screen Test this by increasing decreasing the UV laser power and observing the middle peak Refer to the multi hole target and set the alignment mode for highest hole Move the pin on the bottom of the target to line up with desired target hole and lock the pin down with setscrew Install the highest target in the space you left between the beam translator and the UV dichroic on the optical rail ensuring the target is square to the table top in all respects Rotate the translator window so the UV beam aligns with the top hole in the alignment target Remove the raised UV laser target PN 4237003C 24 25 26 SERVICE AND REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE Use the adjustable UV laser dichroic on the optical rail and look at both traces on the Digiscope screen Adjust the UV beam using the 90 degree UV dichroic only for the same time frame as the HeNe laser generated signal Verify that the time separation between the peak of the UV and HeNe laser generated signals and the Argon laser generated signals is e 40 microsecond
373. illator card processes the delayed sort signal received from the Sort Delay card to provide the fine delay adjust phase When enabled the abort circuitry looks one drop period four quarters ahead and one drop period four quarters behind the quarters containing the desired sort signal to detect possible coincident events The card aborts all non identical coincident events To accomplish this task a short shift register allows the local circuitry to simultaneously examine nine quarter droplets gt One quarter droplet where the event in question lies gt Four quarter droplets ahead gt Four quarter droplets behind PN 4237003 2 8 11 2 INSTRUMENT DESCRIPTION SORT SUBSYSTEM Note For Elite ESP systems abort can be set to eliminate all potentially coincident events The Elite ESP surrounds all detected events with an abort window this includes cells that would and would not be sorted Transforms the sort pulse from TTL type signals to an analog bipolar signal The front and back porch signals are generated in this process Note The users choice of Number of Drops controls the length of the sort pulse This sort signal then goes to the Sort Output card for amplification Three coincident conditions can occur See Table 2 8 1 Table 2 8 1 Coincident Condition Responses Condition Response System Response Explanation Good cell Extension System attempts The logic circuit extends the sort pulse to sort th
374. in 6028009 Cable Lister 6028278 Cable ribbon SCSI host 8 1 24 PN 4237003C Table 8 1 23 Workstation Continued MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY PARTS LISTS Part Number Description Figure Item 6028302 6028304 6028371 6414243 6414379 6414520 6414535 6414852 6414853 6414976 6414977 6604245 6604418 6703473 6703474 6703672 6704278 6704459 6704473 6704556 6704665 6704965 6705318 6705582 6705815 6706418 6855899 6856379 6856736 6856752 6857978 6912111 6912518 6912579 6912612 Cable power module control 25 pin external 10 ft Cable Cytometer 12 10 ft Cable fiber optic Disk REM flex 5 25 in Software Flownet 3 5 in Disk DOS 4 01 5 25 in Disk Flownet Node option Software OPTune 3 5 in Software OPTunee 5 25 in Software Elite 4 00 disk 1 Software Elite 4 00 disk 2 Printer Paintjet Hewlett Packard Card MULTICYCLE Card Extender Profile Card extender driver Profile Card CRT contrast Profile Card motherboard Faraday Profile XT Card Backplane Profile AT Drive extender Flownet 128K Profile Interface DT Profile AT Card Lister Interface DT Profile XT Transputer cytometer TSTD Card Extender XL Software library XL Software library Elite version 4 02 Bracket trigger Profile Lister 88 Profile Card CPU Profile AT CRT 2 Profile Drive Panasonic Profile XT Drive 90 MB B
375. in the Acquisition card cage Connects the Bitmap and Decision card via a ribbon cable to load the sorting bitmaps Gated Amp Control Card Elite systems have this card it provides an interface to the gated amp backplane upper backplane 25 6 42370036 2 INSTRUMENT DESCRIPTION CONTROL AND DISPLAY SUBSYSTEMS VS90E002 gt SICH H3M01 1no 431955 8 1HOS 8 uaisn SOVJHU3INI viva Ir n T HOLIMS 0399079 HALTS ALSVM HO LIIMS 1399019 Y43114 H38IAVHO HOLIMS 3unss3ud WalsAs SENDER HOLIMS ALda 3SNIH HOLIMS Tin 31SVM HOLIMS ALAD H1V3HS GU 1 H3MOd vr xn HOSN3S WOu3 3unssaud 6 oir HLV3HS 17 uo1vinoau sr 3unssdud 3OVJH31NI er HLYJHS P Vo uoivinoau 9 90QH91VM 3H SS3Hd HOHLNOO 3ldWvs aaiv 9 2 SIE H3ddn S 7OHINOO z duvo 89 lt SH313WOILN31Od OML NOILdO SNOTOOLNY HOLON SHOLOW OML IWSINVHO3IN WsINvHo3w uonoeuuoo auva L 03395 NOILISOd VH3IvO 3NO1001nv HANOTOO LNV 158313d 1 HOSN3S
376. ing outwards Install the ribbon cable connector with pin 1 towards the top Markings inside connector Install lower Bertan card cage card retainer Fasten new Bertan 5 power supply HV cable to bracket 5 connector position 3 11 1 INSTALLATION PROCEDURES 5TH PMT KIT WITH SWITCHABLE AMPS INSTALLATION PROCEDURE 3 11 2 10 11 12 13 1 Route the new coax cable the same as the other four HV coax cables and connect one end to Bertan card cage bracket PMT 5 connector position Note The new PMT becomes PMT 4 and the previous PMT 4 becomes PMT 5 Install the new PMT in fourth PMT position from right facing the instrument Lock in position with a 6 32 X 1 2 in thumbscrew a Connect DC Power connector to an open position on the PMT DC Power Distribution card under the HeNe Laser b Connect BNC male end of new cable to new PMT Signal connector 1 Labeleach end of the cable PMT 4 2 Route cable with previously routed PMT signal cables c Connect existing PMT 4 HV cable to newly installed PMT 4 HV connector d Connect newly installed PMT 5 HV cable to old PMT 4 HV connector new PMT 5 Relabel old PMT 4 signal BNC connector as PMT 5 e Remove 3 Signal coax cable from instrument and replace it with new signal coax cable Label both ends PMT 3 and route with existing signal cables Configure 3 PMT Sub Amp Amp SW R2 card as follows Note Solder pin 8 and 9 of U11 Sub SW R
377. ining e Cartons received Power requirements e Water Drainage Environment e Ventilation e Space and accessibility e Labels Supplies Education Center Training Verify that at least one person from the customer laboratory is already trained or is assigned to a class date within two weeks of installation Cartons Received l Compare the cartons received against the customers order and notify shipping if there is a discrepancy 2 Inspect the cartons for damage If any damage exists confirm that a claim was filed with the carrier Power Requirements Power requirements vary for each country It is important that the correct electrical input is available prior to installation WARNING Risk of personal injury Ensure that there is a main power disconnect switch in the same room as the lasers to avoid personal injury l Verify that there are sufficient power outlets available See Table 3 1 1 2 Verify that there is an appropriate power outlet for any accessories including printers and optional disk drives PN 4237003 3 1 1 INSTALLATION PROCEDURES PREINSTALLATION CHECK Table 3 1 1 Power Requirements Omnichrome 56 HeCD Coherent 305 Country System Power Laser Power Laser Water Cooled Laser t USA and other 115 Vac 50 60 Hz 115 Vac 50 60 Hz 115 220 Vac 50 60 208 Vac 10 3 phase with applicable at 20A at 20A Hz Power ground no neutral 50 60 Hz countries Consumption lt 300 W Recom
378. ion Ready 3 10 3 Calibration Procedure 3 10 6 Adjusting Autoclone Sorting Option Speed 3 10 7 Testing the Autoclone Sorting Option Mechanism 3 10 7 Prepare the System 3 10 7 First Test 3 10 8 Second Test 3 10 9 Stream and Waste Catcher Adjustment 3 10 9 Operational Test 3 10 9 5TH PMT KIT WITH SWITCHABLE AMPS INSTALLATION PROCEDURE 3 11 1 Purpose 3 11 1 Tools Supplies Needed 3 11 1 Hardware Configuration and Installation 3 11 1 Cabling Configuration and Installation 3 11 2 Software Installation 3 11 4 System Test 3 11 4 PN 4237003C 3 3 12 BEAM TRANSLATOR UPGRADE 3 12 1 Tools Supplies Needed 3 12 1 Laser Mounting Procedure 3 12 1 Standard Laser Alignment Procedure 3 12 1 Collinear Laser Alignment Fine Tuning Procedure 3 12 4 Time Space Separated Laser Alignment Procedure 3 12 5 3 13 SORT PERFORMANCE ESP UPGRADE 3 13 1 Purpose 3 13 1 Tools Supplies Needed 3 13 1 Preliminary Procedure 3 13 1 Sort Kit Installation 3 13 3 ESP Equipped Units 3 13 5 3 14 ENTERPRISE LASER UPGRADE 3 14 1 Purpose 3 14 1 Tools Supplies Needed 3 14 1 Important Operating Notes 3 14 2 Installation 3 14 2 3 15 COLLECTION OPTICS UPGRADE 3 15 1 Purpose 3 15 1 Tools Supplies Needed 3 15 1 Flow Cell Replacement Procedure 3 15 1 Snout Upgrade Procedure 3 15 1 Purpose 3 15 1 Tools Supplies Needed 3 15 1 Removal Procedure 3 15 2 Replacement Procedure 3 15 3 3 16 PIEZOELECTRIC TRANSDUCER I
379. irror reflects 488 nm 7 3814222 633 mirror reflects 633 nm 8 N A Laser beam 9 6857236 Small target 10 6858459 3 hole multipositional target 8 2 36 PN 4237003C Figure 8 2 37 Cytometer Inside Right Rear View See Table 8 2 37 ee e ee ee ee e Wi Table 8 2 37 Cytometer Inside Rear View Detail of Parts See Figure 8 2 37 SNe d A ILLUSTRATED PARTS EN 7003136A PARTS LISTS Item Part Number Description 1 6857604 Cover top removable 2 2603044 Fan blower 200 CFM 115 Vac 3 6028016 Cable laser controller 4 6857637 Laser interlock relay bracket 5 6704933 Cytometer card cage 6 6856911 Transformer drawer PN 4237003C 8 2 37 8 PARTS LISTS ILLUSTRATED PARTS 7003141A Figure 8 2 38 Omnichrome HeCd Laser See Table 8 2 38 1 Table 8 2 38 Omnichrome Laser See Figure 8 2 38 Item Part Number Description 1 6912834 HeCd 74 laser Note Use PN 3814244 for head and power supply 325 nm 30 mW 2 6028256 Interlock cable 3 6027225 Line cord 3 18 ga 8 2 38 PN 4237003C QUICK REFERENCE INFORMATION A 1 1 1 TOLERANCES AND LIMITS A 1 1 Pressure 1 1 Compressor Module A 1 1 House Air 1 1 Pneumatics 1 1 Switches A 1 1 Initial Installation Settings A 1 1 Argon Laser A 1 1 Camera 1 1 Front and Back Porch Settings for Best Streams A 1 1 Test Points A 1 2 Pneumatic I
380. is visible just above the ground plate so that you can easily monitor it Phase Adjustment 1 Set the Sort Counters to OFF 2 Select a Sort Test b Left Sort and Right Sort c High Voltage PN 4237003C 3 6 3 INSTALLATION PROCEDURES SYSTEM TESTING 3 6 4 3 Setthe Sort Droplet to 3 and press START to activate the sort counters Sort counters should be running IMPORTANT Risk of incorrect results If the side streams hit the sort plates incorrect results may occur Do not allow the side streams to hit the sort plates 4 Increase the deflection by increasing the Deflection amplitude to bring out the side streams 5 Find the best side streams by using the phase adjustment which is the decimal portion of the Delay setting You should get at least seven of the phase settings to provide good clean side streams If not repeat step 4 Note The problem may be an incorrect Drive setting Optimal side streams are evident when the filament between the two drops is very thin and remains connected to the drops 6 Find the worst phase setting and subtract 0 5 Set the phase to that number to provide the best side streams Deselect Sort Test to turn off Load the sample protocol created in step 1 of Acquisition Check Setthe Sort Settings to Receive 10 Send the Cyto Settings 11 Change the Sort Settings to Send to send your sample sort gates IMPORTANT Risk of compromising sort stability if you increase the sampl
381. is when a new flow cell flow body or bimorph stage is replaced 5 Ifthe stream is still too close to one side or the other verify through proper alignment procedures that the Z axis of the flow cell stage is adjusted correctly Any movement of this axis can affect the rest of the optical alignment Drive Frequency Sort Settings l Setthe Drive Frequency at 32 kHz 2 Set the Crystal Drive to 7596 4 2 4 PN 4237003C PN 42370036 SERVICE AND REPAIR PROCEDURES SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE While viewing the droplets above the ground plate scan through the frequency settings using the tenth decimal adjustment Note It is best to start at 32 kHz and scan down Stop scanning when the highest breakoff closest to the flow cell is achieved and the droplets appear very focused and sharp Set a cursor tangent to the bottom of the last attached drop Ifthe last attached droplet is not yet visible above the top of the ground plate increase the Crystal Drive from 7596 to 8596 Ifthe last attached drop is still not visible move the deflection body down slightly e Adjust the drive for a thin filament connecting the last attached drop Press DEBUBBLE twice with High Voltage off and ensure that the breakoff returns to the same place If the breakoff does not return to the same place the flow cell has a bubble or a partial clog Remedy this before continuing In extreme cases you may have to remove or r
382. ise Amplitude drift Warm up time Beam pointing stability Lifetime Operating voltage Operating current 250 1 minimum 1 5 RMS 10 Hz 2 MHz 1 maximum light control mode over 2 hours 5 minutes maximum lt 0 03 mrad after warm up temperature range 3 C gt 10 000 hours at specified power 115 Vac 10 17 0 A maximum single phase Uniphase Red Output power 10 mW minimum HeNet Wavelength 632 8 nm Polarization Random Spatial mode TEM oo Beam diameter at 1 e2 0 68 0 02 mm Beam divergence 1 2 mrad Amplitude noise 0 5 RMS 30 Hz 10 MHz Amplitude drift 3 Warm up time 15 minutes maximum Beam pointing stability 0 02 mrad maximum after warm up Lifetime 10 000 hours minimum Operating voltage 3100 200 Vdc Operating current 6 5 0 2 mA Melles Griot Minimum power 1 5 mW Green Hehe Wavelength 543 5 nm Mode Polarization Random Beam diameter at t e2 Beam divergence Amplitude drift Amplitude noise Warm up time Beam pointing stability Operating temperature Expected tube life Line frequency Operating voltage Operating current 0 80 5 mm 0 86 5 mrad lt 2 5 lt 2 8 30 Hz 30 MHz peak to peak 7 minutes lt 0 03 mrad after 15 minutes 20 to 40 C 8 000 hours 48 63 Hz 3 000 100 Vdc 6 5 0 2 mA Due to low output power this laser is not recommended for sense in air operation tAir cooled laser dWater cooled laser PN 4237003C 2 3 3
383. itchable 4 PMT Switchable Amps with Peak Amps Scatter Function t Earliest configuration First field upgradable Factory configuration available from configuration February 1992 3 PMT Sub Compensates the FL1 FL2 Compensates combinations Compensates the FL1 FL2 and FL3 SW R1 card and FL3 PMT signals of the FL1 FL2 and FL3 PMT signals PMT signals 3 PMT Sub Not present Compensates combinations Not present SW R2 card of the FL2 FL3 and FL4 PMT signals The system cannot compensate for an F1 and F4 combination because they are not routed to a common card This is not a problem however because the two PMTs normally use optical filters with passbands that are far enough apart to prevent any spectral overlap peak scatter sensor is the FALS sensor in this system and is also used in the Gated Amp system 2 6 8 PN 4237003C Table 2 6 2 Configuration Summary Continued INSTRUMENT DESCRIPTION ACQUISITION SUBSYSTEM FL1 Routed to the Dual FL Amp Routed to 3 Sub Routed to the Dual FL Amp card for card for linear and log SW R1 card for linear and log amplification amplification compensation FL2 Routed to the Dual FL Amp Routed to both PMT Sub Routed to the Dual FL Amp card for card for linear and log SW R cards for linear and log amplification amplification compensation FL3 Completed on 3 PMT Sub Routed to both PMT Sub Completed on PMT Sub SW R card SW R
384. ium 74 Laser Figure 1 2 5 Laser Warning Labels Coherent Innova Enterprise Laser Figure 1 2 6 Laser Warning Labels Coherent Innova 305 Laser Figure 1 2 7 Laser Warning Labels Coherent Spectrum Laser Figure 1 2 8 Laser Warning Labels Uniphase Red Hehe Laser e Figure 1 2 9 Laser Warning Labels Melles Griot Green Hehe Laser Figure 1 2 10 Laser Warning Labels Sensing Area Figure 1 2 1 Laser Warning Labels Coherent Innova 300 Water Cooled Argon Laser 2WARNING DANGEROUS VOLTAGE UNDER THIS COVER VISIBLE AND INVISIBLE RADIATION WHEN OPEN AND INTERLOCK DEFEATED AVOID EYE OR SKIN EXPOSURE TO DIRECT OR SCATTERED RADIATION VISIBLE AND INVISIBLE LASER RADIATION RASER R AVOID EXPOSURE FROM THIS APERTURE VISIBLE AND INVISIBLE LASER RADIATION DANGER AVOID EYE OR SKIN EXPOSURE TO VISIBLE AND INVISIBLE LASER DIRECT OR SCATTERED RADIATION RADIATION WHEN OPEN AND INTERLOCK DEFEATED AVOID EYE OR 12 WATTS MAX GW SKIN EXPOSURE TO DIRECT OR SCATTERED RADIATION PRACTICAL LIMIT CLASS IV LASER PRODUCT 5965007B 1 2 3 1 INTRODUCTION SAFETY PRECAUTIONS Figure 1 2 2 Laser Warning Labels Innova 90 Water Cooled Argon Laser A INNOVA 90 ARGON LASER VISIBLE AND INVISIBLE LASER RADIATION AVOID EXPOSURE FROM THIS APERTURE VISIBLE AND INVISIBLE LASER RADIATION DAN G E R AVOID EYE OR SKIN EXPOSURE TO LASER RADIATION WHEN OPEN DIRECT OR SCATTERED RADIATION AND INTERLOC
385. k Ground J3 Blue 2 3 to 3 0 Vdc J4 Red 2 3 to 3 0 Vdc Optics Beam Expander Size of beam entering beam expander approximately 0 3 mm in diameter Size of beam exiting the beam expander approximately 0 15 mm in diameter before passing through the HeNe mirror Signal Adjustments Beam Translator Upgrade Adjust between the peak of the Argon caused signal and the peak of the UV caused signal 20 microseconds for a 100 p tip 30 microseconds for a 76 p tip Adjust between the peak of the UV and HeNe and the Argon caused signals for time separation of 40 microseconds for the 100 p sort sense tip at 12 psi or 60 microseconds for the 76 sort sense tip at 12 psi or 7 microseconds for the 76 jet in air flow cell tip A 1 2 PN 4237003C QUICK REFERENCE INFORMATION A TOLERANCES AND LIMITS Gated Amp Laser Alignment Time Space Separated Laser Alignment Adjust between the peak of the HeNe laser generated signal and the peak of the UV laser generated signal by adjusting the HeNe laser dichroic on the optical rail 20 microseconds for a 100 tip 30 microseconds for 76 tip Adjust between the peak of the Argon laser generated signal and the peak of the UV laser generated signal by adjusting the Argon dichroic on the optical rail 20 microseconds for a 100 p tip 30 microseconds for a 76 p tip Time separation between the peak of the UV and HeNe laser generated signals and the Argon laser generated signals
386. k known as the Driver circuit Note Only the Pneumatic Interface card circuitry totally controls the regulation process Once the CPU and Sensor I O cards provide a value to the Pneumatic Interface card their roles end The system regulates sample pressure by comparing the DAC voltage that indicates the desired pressure with the voltage that the Sample Pressure Sensor circuit develops After amplification the difference known as error voltage drives the electronic pressure regulator R51 controls the power supply for the sample pressure sensor This is nominally set to provide 8 00 V at TP2 which is the voltage input to the sensor The sensors output depends on this voltage and on the pressure input You can monitor the sensor output at TP7 Note This is also the voltage the system applies to the Sensor Interface card for the readback display Although the sensor is linear there is a 1 V offset from zero The circuit compensates for this by summing a 1 V level with the sensor output to produce the feedback voltage applied to the op amp driver circuit for the regulator You can adjust the offset compensation by R59 Therefore R59 affects the Low Pressure circuit response more than it affects the High Pressure circuit response and the DAC reference voltage R34 affects the High Pressure circuit response more than it affects the Low Pressure circuit response The adjustment procedure for the Pneumatic Interface card balances these two adju
387. king Beam Spot Dimensions 2 3 7 CRI Display 2 5 4 Dye Emissions 2 6 6 Configuration Summary 2 6 8 Typical Applications of Beam Shapers 2 7 6 Coincident Condition Responses 2 8 12 Power Supply Functions 2 8 19 Solenoid Valves Manifold 2 9 2 PN 4237003C INSTRUMENT DESCRIPTION 2 1 INSTRUMENT OVERVIEW Function The Elite is a cytometry analyzer that processes flow cytometric measurement and sorting of cells within a sample suspension For more information on the Elite s function refer to Chapters 1 and 3 of the Reference manual Description Main Components Two main components comprise the Elite the Cytometer and the Workstation See Figure 2 1 1 Figure 2 1 1 Elite Components COMPUTER WORKSTATION CYTOMETER mmm mmm SCOPE TOUCH DETECTOR SENSOR SCREEN SCREEN COMPARTMENT MODULE COLOR MONITOR Kees SORT COLLECTION INTRODUCTION COMPARTMENT i ST us SHEATH CABINET CPU ELECTRONIC POWER AND WASTE PEDESTALS SWITCHES COMPARTMENT Subsystems The Elite is comprised of several subsystems each of which is described in detail later in this manual The subsystems include e Control and Display subsystems see Heading 2 5 gt Multibus subsystem see Multibus Subsystem under Heading 2 5 gt Cytometer Computer subsystem see Cytometer Computer Subsystem under Heading 2 5 PN 42370036 21 1 INSTRUMENT DESCRIPTION INSTRUMENT OVERVIEW Laser Control
388. ks or registered trademarks of their respective holders PN 4237003C
389. l s composition The Elite correlates and stores several measurements of cell characteristics and graphically displays the distribution of those characteristics within the sample The system can then sort the cells based on the measured characteristics Sensor Area The Sensor area Figure 2 1 2 consists of the Sample introduction system Sensing compartment e Sort collection area e Detector compartment e Graphics and touch screen monitors PN 4237003C INSTRUMENT DESCRIPTION 2 INSTRUMENT OVERVIEW Figure 2 1 2 Sensor Area GRAPHICS TOUCH SCREEN DETECTOR SENSING MONITOR MONITOR COMPARTMENT COMPARTMENT 000 005 BH 59040348 SORT INTRODUCTION COLLECTION SYSTEM AREA Sample Introduction System The sample stage Figure 2 1 3 accepts samples for analysis in 12 x 75 mm test tubes Sample pickup is by positive pressure Sample flow is guided into the sheath stream at the flow chamber Sheath fluid flows from a sheath tank in the sheath and waste compartment An internal compressor pressurizes the sheath There is a bank of controls to the right of the sample stage that provides manual control of sheath and sample flow Figure 2 1 3 Sample Stage 7003110A PN 4237003C INSTRUMENT DESCRIPTION INSTRUMENT OVERVIEW 2 1 4 PN 4237003C INSTRUMENT DESCRIPTION 2 AUTOCLONE SORTING OPTION 2 2 AUTOCLONES SORTING OPTION General The Autoclone Sorting Option Figure 2 2 1 is a
390. l FL SW R card for linear and log amplification e The 3 PMT Sub Amp card completes processing the FL3 signal e Coax cables take all processed signals to the Mux and Scope card This configuration uses the peak scatter FALS sensor e Coax cables carry the FALS signal to the Peak Scatter Mux SW R card The Peak Scatter Mux SW R card splits the signal in two One signal is routed to J12 and then to the Scat CV SW R card which processes the signal to provide the integral and log FALS signals The second signal leaves the Peak Scatter Mux SW R card from J14 where coax cables carry the signal directly to the Mux and Scope card PN 4237003 2 6 11 INSTRUMENT DESCRIPTION ACQUISITION SUBSYSTEM VGE0 00 eH oir eld 110 23 13 8 H9 901 21 1010 93 63 17 DOFELNd or D 170 73 63 INI 214 5 110 53 13 INCEIN ir sr 100 2714 9 jno zu NI LH OHO MIN if 100 14 980 t j 907 9 110 74 Pr FH lt INFZIWd er 1NI E13 4 4 907 674 DHI 504 27 IECH MEIH 8 oz SO TFIWG NEU 9n L A WHO eir INFFIWd Y MS ANS et
391. l flow summary of 2 6 9 FL4 signal flow summary of 2 6 9 flow cell alignment procedure 4 1 3 defective determining if 4 11 4 illustration removal of shipping plug 3 5 2 replacement procedure 4 11 4 sort sense adjustment procedure 4 11 1 sort sense verification procedure 4 11 1 tips See flow cell tips verification procedure 4 11 4 flow cell tips high sensitivity description of 2 7 8 information on upgrading 3 15 1 standards of September 1993 and beyond 3 15 1 upgrading the snout 3 15 1 flow cell vacuum valve VL5 function of 2 9 2 flow cells bodies old and new 4 12 3 flowcharts fluidics troubleshooting 7 3 3 troubleshooting sort problems 7 5 1 fluidics operational test See VIP fluidics See pneumatics INDEX 7 INDEX fluorescence pickup lens location 4 12 9 focus knob location 4 12 11 focusing the camera See camera forward scatter forward scatter detector 2 3 13 function of lens 2 3 13 light description 2 3 13 obscuration templates 2 3 13 forward scatter detector mask laser beam centered in 4 12 6 functions See specific entries G gate window defined 2 12 2 gate open collector function of 2 13 4 Gated Amp AUX 1 adjusting 3 9 16 AUX 2 adjusting 3 9 16 calibrating the laser 3 9 24 cell passing through laser beam description of 2 6 8 description 2 6 6 3 9 1 function 2 6 6 2 6 8 installation procedure 3 9 2 installation requirements 3 9 1 laser alignment procedu
392. l injury The laser beam can cause eye damage if viewed either directly or indirectly from reflective surfaces such as a mirror or shiny metal surface To prevent eye damage avoid direct exposure to the beam Do not view it directly or with optical instruments except with special service tools as directed in this manual 3 7 1 INSTALLATION PROCEDURES OPTIONAL LASER INSTALLATION 3 7 2 1 Determine which laser kits are present Kit Part Number Cover Upgrade 6912833 Optics 6912834 Laser Mounting 6912844 for Innova 305 laser 6912841 for 90 5 laser Water cooled 305 laser 6912831 HeCd laser 6912801 Water cooled 90 5 laser 6912832 CAUTION Risk of instrument damage The installation procedures shipped with the kits may contain the following errors e Installation Procedure Optics PN 9022633 Rev A gt Item 5 should be PN 6857612 633 nm mirror gt Item 4 should be PN 6857611 488 nm mirror e Installation Procedure HeCd Model Kit PN 9022632 Rev A gt Item 2 should be PN 2814049 1 4 20 x 9 16 screw gt Item 11 should be PN 6857611 gt Ext Interlock Cable PN 6028256 is needed but not shown Examine the packing list for each kit to ensure that all parts are present and notify shipping of any discrepancy Note installation procedure should accompany all kits Inspect all optional lasers for correct operation For specifications and procedures refer to the manual s supplied with the laser
393. l to be sorted is in the correct place This function is complicated because cells appear in the laser beam at random times while a droplet can only be sorted at the time it breaks off from the end of the stream Therefore the randomly appearing sort requests must be delayed and synchronized with the droplet formation rate Receives input from the Bitmap and Sort Decision card indicating an event should be sorted right or left The card synchronizes the inputs to the digital clock signals received from the Sort Oscillator card These clock signals are synchronous with the droplet formation rate but at a higher frequency the f20 clock Delays the synchronized sort signals in a type of shift register for the user selected number of droplet periods The key is that the card uses a clock frequency that is synchronous with the droplet formation rate The card samples the sort signals and loads them into the delay memory at four times the formation rate This provides the system with a quarter drop sort resolution Sends the delayed sort signals to the Sort Oscillator card PN 4237003C 2 INSTRUMENT DESCRIPTION SORT SUBSYSTEM 2 8 9 050500 ENEE 03 939 9j8J ay soun p 1e 301111950 SA Sajejs 9 ufnouu Sa 1908 1012 9199
394. lect Streched Pulses backplane and P1 to be displayed connector 7003070A Quad Peak Sense and Hold PSH Cards 2 Together two Quad PSH cards Figure 2 13 2 provide eight parallel acquisition channels Each channel s purpose is to develop a voltage output that represents the maximum value peak of the input pulse This voltage output is held by the Peak Sense and Hold circuit until the conversation cycle is completed and the card receives a reset command The discriminator circuitry determines which pulses the card captures and holds Each Quad PSH card is organized into four identical channels The system traces the signal flow through one channel The card Limits incoming signal to 10 V to ensure that all off scale pulses are placed in the last channel of the histogram PN 4237003 2 13 3 INSTRUMENT DESCRIPTION SIGNAL SELECTION AND DIGITIZATION 2 13 4 Implements the discriminator function of the system Before acquisition the system loads a discriminator value into a latch on the card A digital to analog converter DAC converts the latch output to an analog voltage and provides one input to a voltage comparator The input pulses are provided to the other comparator input This comparators output is gated by whether the particular PSH channel is active This condition is set if a discriminator value is set to this channel including channel 1023 if this channel is set to OFF then the output is blocked The card uses
395. leed Regulator Adjustment Procedure PN 4237003C ATTENTION Do this procedure for low bleed regulator instruments only Purpose Do this procedure after the Pneumatic Interface card either regulator or the sheath pressure sensor is replaced The following definitions appear in this procedure Set pressure means enter the needed pressure on the Cytometer Control Valves screen Gauge pressure refers to the pressure read on the external test pressure gauge Read pressure refers to the pressure displayed on the Cytometer Control Valves screen Tools Supplies Needed C DVM Digital pressure gauge 0 01 psi accuracy Pneumatic Interface Card Adjustment l Access the Pneumatic Interface card a Remove the rinse bottle b Remove the rinse bottle mounting bracket c Remove the Pneumatic Interface card splash shield 2 Connect the negative DVM test lead to the lower lead of C11 marked on the Pneumatic Interface card 3 Place the positive DVM lead on the test points indicated in Table 4 5 1 and adjust the appropriate potentiometer for the indicated voltage IMPORTANT Risk of erroneous results if the voltage adjustments are made in the wrong order Adjust the potentiometers for the indicated voltages in the order shown in Table 4 5 1 Table 4 5 1 Potentiometer Adjustments Adjust For At Test Point R31 10 00 0 01 Vdc TP5 R34 10 00 0 01 Vdc 1 R81 10 00 0 01 Vdc TP10 R54 10 00 0 01
396. let sort the cells abort if they are closer than four droplets apart This permits a bad cell in the first droplet to be detected resulting in the system aborting both cells Definitions Good a particle or cell that meets the conditions to be sorted meets the sort equation by being in the correct regions and or bitmaps Bad a particle or cell that does not meet the conditions to be sorted Direction left or right Droplet as a measure of time the reciprocal of the droplet frequency gives the period of one droplet At 32 KHz one droplet period is 1 32 KHz therefore 1 32 KHz 31 microseconds This allows you to relate the time difference between pulses as observed on the scope relative to how far apart they will be in the sort stream in droplets Followed by implies the second cell is close enough to the first cell to create a sorting problem Sort Output Card The Sort Output card Figure 2 8 8 PN 4237003C Amplifies the crystal drive and sort pulse signals to appropriate voltage levels Controls the amplitude of these signals to provide the crystal drive and sort deflection controls Buffers the FL test and strobe signals from the Sort Oscillator card to provide outputs suitable to drive LEDs Adjusts the FL test signal s amplitude based on operator input 2 8 13 2 INSTRUMENT DESCRIPTION SORT SUBSYSTEM Figure 2 8 8 Block Diagram Sort Output Card and Transistor Bracket Card FROM CPU SORT PULSE
397. level to the Sensor Interface card indicating sample pressure This is provided for display only and does not affect the sample pressure control circuit Figure 2 9 3 Block Diagram Sample Pressure System New Type VOLT REF R31 FROM CPU 7003076 5 1 VOLT REF R34 INPUT LATCH O DAC 0 10VIN 15V V Regarding Figure 2 9 3 The Input Latch is an 8 bit word set pressure written by CPU U23 SENSOR OUT 8 00 V TP7 TP2 SENSOR T 1 1 1 1 BUFFER REF VOLT 1 51 J3 TO SENSOR 1 0 1 t SAMPLE REGULATOR 1 1 SENSOR l aT E gt amp 3 15 PSI DRIVER OUT 30 PSI IN The DAC provides a voltage proportional to the digital pressure Full scale is set by Volt Ref 2 9 7 2 INSTRUMENT DESCRIPTION PNEUMATICS SUBSYSTEM 2 9 8 Sheath Pressure Systems There are two types of sheath pressure systems old type and new type Old Type With the old type of Sheath Pressure system Figure 2 9 4 the Sheath Pressure Regulation circuit is similar to the old type Sample Pressure circuit in that the CPU writes the desired pressure to the Pneumatic Interface card The card compares the desired pressure with the pressure measured by the sheath pressure sensor The Pneumatic Interface card t
398. ll body Figure 4 12 4 1 Loosen the top knob 2 Remove the knob and insertion rod 3 Loosen the bottom knob slightly 4 Rotate the flow chamber until the beam reflects back onto itself 5 Tighten the knob 6 Hold the flow cell in place and replace the insertion rod 7 Tighten the top knob Figure 4 12 4 Flow Cell Bodies Old and Newv OLD TOP KNOB L e 7003206 b Fora new flow cell body Figure 4 12 4 1 Loosen the knob slightly 2 Rotate the flow cell chamber until the laser beam reflects back onto itself 3 Tighten the knob 4 Replace the beam shaper and tighten the screw ATTENTION The newer flow cell tips Figure 4 12 5 appear slightly lower when in the correct position than do the older flow cell tips Figure 4 12 6 6 Place the tip at the center of the fluorescence pickup lens a Turn the flow cell horizontal adjustment as needed b Turn the flow cell vertical adjustment as needed Note If the LED is turned on and the mirror is in place a white dot appears as shown in the newer flow cell tip in Figure 4 12 5 If not adjust the flow cell horizontal and flow cell vertical knobs See Parabolic Mirror on Tip Optional for more information PN 4237003C 4 12 3 SERVICE AND REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE Figure 4 12 5 New Flow Cell Body 7003194A Figure 4 12 6 Old Flow Cell Body COLORED DOT 7003195A 7 Do Laser Beam Alignment Parab
399. lock Diagram Sheath Pressure System Old Type 2 9 9 2 9 5 Block Diagram Sheath Pressure System New 2 9 11 2 9 6 Block Diagram Compressor Module 2 9 12 2 10 1 3 PMT Sub SW R Card Block Diagram 2 10 1 2 10 2 Dual FL SW R Card Block Diagram 2 10 2 2 10 3 Scat CV SW R Card Block Diagram 2 10 3 2 10 4 Block Diagram Peak Scatter Mux SW R Card 2 10 4 2 11 1 Block Diagram Scat CV Amp Card 2 11 1 2 11 2 Block Diagram 3 PMT Sub Amp Card 2 11 2 2 11 3 Block Diagram Dual FL Amp Card 2 11 3 2 12 1 Block Diagram Gated Amp Control Card 2 12 1 PN 4237003 2 V 2 vi 2 12 2 2 123 2 12 4 2 12 5 2 13 1 2 13 2 2 133 2 13 4 2 13 5 2 13 6 2 13 7 2 13 8 2 13 9 TABLES 2 2 1 2 3 1 2 3 2 2 3 3 2 5 1 2 6 1 2 6 2 2 7 1 2 8 1 2 8 2 2 9 1 Block Diagram Scat Aux Gated Amp Card 2 12 3 Block Diagram PMT Gated Amp Card 2 12 5 Block Diagram 20 40 60 Microsecond Delay Card 2 12 6 Block Diagram 7 Microsecond Delay Card 2 12 6 Block Diagram and Scope Card 2 13 3 Block Diagram PSH Card s 2 13 5 Block Diagram ADC and PSH Control Card 2 13 6 Block Diagram Peak ADC PSH Control Card 2 13 7 ASD Settings Diagram 2 13 9 Peak ADC PSH Card Timing 2 13 10 ASD Settings Diagram 2 13 11 Old ADC Timing Diagram 2 13 12 Block Diagram Lister Cards 2 13 13 Autoclone Sorting Option Screen Marker Definitions 2 2 3 Laser Summary 2 3 2 Laser Technical Specifications 2 3 3 Wor
400. logic e front back porch e sort clip cycle in PURGE Sort pulse shape as sort gates deflection function of drops discsat ext e delay e flowcell mounting liquid leaks bandwidth flowcell position front porch backporch beamshaper Ld bimorph e no leakage current D beam lockout from sort offset e delay timing e coincidence modes e flow body e coincidence operation flow cell tip all modes 7003083A Tools Supplies Needed DVM Sort Current Leakage Test Put system in Sheath Ensure Sort Test Sort Right and Left Enable are off Use your DVM to measure voltage on the sample insertion rod of the flow cell body gv a Adjust Stream Offset and Deflection percent for maximum voltage You should read at least 80 V Reduce Deflection percent to 0 0 Remove the alligator clip from sample intro rod and connect to one of your meter leads Connect the other meter lead to the sample intro rod Set your meter to measure mA Slowly increase the deflection percent while watching the meter s display Ifmore than 1 mA of current is detected there is an unwanted current path from the sheath liquid to ground The problem can be a cracked fitting or a metal bulkhead fitting all tubing carrying fluids that connect to the flow cell must go through plastic feed thru fittings 7 5 1 PN 4237003C 7 TROUBLESHOOTING TROUBLESHOOTING THE SORT SUBSYSTEM Table 7 5 1 Troubleshooting Sort Problems
401. loscope You will see the normal 3 drop sort envelope N o MW 9 Slowly adjust R109 to reduce the pulse delay while observing the sort pulse shape 10 At about 90 microseconds the three drop envelope shorten into two drop envelope At about 60 microseconds the envelope shortens into a one drop envelope this verifies operation of the truncation function Extended Abort Test This procedure applies to ESP equipped systems only and allows the system to look ahead of a good cell and abort if a bad cell is too close Adjust R109 for a 100 microsecond delay Turn Sort Left Enable ON and Sort Right Enable OFF Turn Abort to OFF and readjust your oscilloscope to observe the sort pulses Turn Coinc Abort to ON Select Ext Sort mode While observing the sort pulse adjust R109 to reduce the delay until the sorting stops Note the delay in microseconds zt ze Wc D Select Comp Abrt mode Slowly increase the pulse delay with R109 until sort pulses reappear Note the delay in microseconds 8 Verify that the delay obtained in step 7 is greater than that obtained in step 5 to verify operation of the Extended Abort mode See Figure 4 2 3 PN 4237003 4 2 13 SERVICE AND REPAIR PROCEDURES SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE Figure 4 2 3 Pulse Signals and Histograms for Sort Coincidence Verification The first PMT 2 INT pulse being larger forms this population The second 2 INT pulse being sm
402. ltibus Subsystem 2 5 1 Cytometer Computer Subsystem 2 5 1 Multibus CPU 2 5 1 External Memory or 256K Memory Card 2 5 2 Serial I O Card 2 5 2 Laser Control Subsystem 2 5 2 Dual Laser Control Card 2 5 2 Video Subsystem Cytometer 2 5 4 Camera Image Signal Flow Video Display 2 5 5 Camera Interface Card 2 5 5 Dual CRT Control Card 2 5 5 Digiscope Card 2 5 5 Pulse Video Display 2 5 5 Cytometer Control Subsystem 2 5 6 Description 2 5 6 Data Taker Interface Card 2 5 6 Gated Amp Control Card 2 5 6 Sensor I O Card 2 5 8 Motor Control Card 2 5 8 DAC Card 2 5 8 ACQUISITION SUBSYSTEM 2 6 1 Analog Signal Processing 2 6 1 Subtraction Color Compensation 2 6 1 Log Amplification 2 6 2 Integration 2 6 4 Special Functions 2 6 5 Switchable Amplifiers Bandwidth 2 6 5 Gated Amp Concept 2 6 6 Signal Flow for Each Configuration 2 6 8 4 PMTs Nonswitchable Amps 2 6 9 4PMTs Switchable Amps with Peak Scatter Function 2 6 11 5PMTs Nonswitchable Amps No Gated Amp 2 6 13 5PMTs Switchable Amps No Gated Amp 2 6 15 Gated Amp 2 6 17 OPTICAL ELEMENTS 2 7 1 Overview 2 7 1 Light 2 7 1 Definition 2 7 1 Media Used for Travel 2 7 1 Snell s Law 2 7 2 Lenses 2 7 2 Positive Lens 2 7 2 Negative Lens 2 7 3 Equations 2 7 3 4237003C PN 42370036 2 8 2 0 2 10 2 11 CONTENTS 2 Optical Components Used in the Elite 2 7 4 Beam Shapers 2 7 5 Beam Expander Reducer 2 7 7 Parallel Plate 2 7 7
403. m LP 3814191 3814266 6858371 1 0 ND filter Elite ESP 3802048 6859060 610 nm BP filter Elite ESP 3814273 6859061 640 nm DL Elite ESP assembly 3814137 8 1 10 PN 4237003C MASTER PARTS LIST IN NUMERICAL ORDER CATEGORY Table 8 1 8 Flow Cell Area PARTS LISTS Part Number Description Figure Item 1019009 Flow cell holder Elite Analyzer Bracket holder flow cell mount 8 2 20 5 1019539 Bracket interlock Elite Analyzer 8 2 20 1 1010933 Clamp nut Elite Analyzer 8 2 20 7 1016487 Tubing sample 7 0 in Elite Analyzer 8 2 20 12 1020903 Bracket tube holder Elite Analyzer 8 2 20 15 2851558 Screw machine 25 20 x 1 0 Elite Analyzer 8 2 20 3 11 3210004 Tubing silicone sample 0 010 i d Elite Analyzer 8 2 17 3 8 2 19 11 8 2 20 6 3814172 Stage flow cell Elite Analyzer 8 2 19 2 8 2 20 14 3814173 Stage beam Elite Analyzer 8 2 19 3 8 2 20 16 6211015 Valve actuator pilot 8 2 5 8 8 2 18 14 23 8 2 19 1 6602642 Flow cell Biosense 8 2 32 5 6704311 Deflection body Autoclone Sorting Option 8 2 19 7 8 2 26 3 6704754 Deflection body standard 8 2 19 7 8 2 21 1 6705182 Sensor peak scatter 8 2 19 9 6705402 Sensor scatter R2 8 2 19 9 6705597 Sensor scatter Elite Analyzer 8 2 19 9 8 2 20 8 6856101 Flow cell tip 100 micron with thin flow body 6856191 Flow cell tip Elite Analyzer 8 2 20 9 6856511 Flow cell tip 76 micron Sort Sense 6856696 Beam shaper 15x60 mm 8 2 19 5 6856826 Beam shaper 8 x
404. m to run in sheath again for the same amount of time as in step d g Fillthesheath tank with Isoflow and leave system in sheath for 5 minutes to get the Isoflow sheath fluid throughout the system h Putthe system in vacuum i Do Flow Cell Replacement and Verification j Remove the biohazard flow cell and the flow body k Install the new flow body and the new sort sense flow cell that the customer will be using 1 Install the new 0 2 m sheath filter and ensure that it fills immediately with sheath m Release any air bubbles in the filter using the sheath purge valve Flow Cell Replacement and Verification Consider a flow cell defective and fail it only when There is discernible fluidics problem in the system Stable breakoff cannot be achieved The droplets appear to constantly drift up and down stream Seven or eight singular good side streams cannot be attained out of ten The flow cell has been tightened sufficiently CLEAR and or DEBUBBLE does not alleviate the instability in the droplets There are no bubbles in the flow cell The correct fittings in the pneumatics manifold are PVC and there is no electrolysis occurring at the point that the insertion rod enters the flow body Electrolysis is evidenced by a buildup of a blackish greenish cloudy substance Verify that there are no metal barbed fittings on the valve bracket assembly and that there is not a blackish greenish buildup where the inse
405. mended capacity 60 A per phase Current consumed 50 A per phase Europe and other applicable countries 220 Vac 20 60 Hz at 10A at 10A 220 Vac 50 60 Hz Optional laser T The electrical service for water cooled lasers may be either WYE or Delta connected with the fourth wire to the building s ground WYE is recommended because it is more tolerant to line voltage imbalances from each phase to ground Water system with an optional Coherent Model 305 Laser requires a source of water that meets the specifications in Table 3 1 2 Table 3 1 2 Water Specifications for Coherent 305 Laser Description Specification Flow rate per minute Minimum 8 5 liters 2 2 gallons Maximum 11 6 liters 3 0 gallons recommended Static pressure Note Static pressure is the inlet pressure measured under conditions of zero flow Maximum 620 kPa 90 psi System CV 0 47 Pressure Delta Note Pressure Delta is the pressure differential between inlet and drain 152 276 kPa 22 40 psi Inlet temperature 10 32 C 50 90 F Temperature stability 1 C 1 8 F Resistivity 5 0 KO cm pH 6 8 Hardness 100 mg l 5 9 grains gallon or 100 parts million of calcium Particulate size 200 y diameter Coherent supplies a 60 p water filter with the laser Total heat load 20 kW at 230 Vac 18 kW at 208 Vac PN 4237003C INSTALLATION PROC
406. ment Procedure This alignment procedure assumes alignment of the Argon laser on top and the HeNe and UV lasers on the bottom PN 4237003C 3 12 1 INSTALLATION PROCEDURES BEAM TRANSLATOR UPGRADE ATTENTION The order in which you align the lasers may need to be changed If a dichroic blocks the light from a laser further out on the optical bench do the dichroic alignment for the interfering dichroic last For example if a 488 nm laser in position three is blocked by the 488 dichroic in position two align the third dichroic before you align the second dichroic Figure 3 12 1 shows the laser beam hitting the dichroic Figure 3 12 1 Laser Beam Hitting Dichroic Top View LASER BEAM DICHROIC TOP VIEW 7003207A l Determine which laser beams particles will pass through first and use that laser as a trigger source for the Gated Amp 2 Determine which lasers are to be used and in what time space dimension top beam or bottom beam 3 12 2 PN 4237003 PN 42370036 INSTALLATION PROCEDURES BEAM TRANSLATOR UPGRADE 3 Install the two 5 in targets on the optical bench in front of the HeNe laser separated by two mounting holes a Remove the dichroics b Install the 5 in targets in front of the first laser HeNe HeCd or other c Align the laser beam through the 5 in hole d Repeat steps a through c for additional lasers Note Allow enough room to see the alignment holes in the targets
407. monitor at all zoom settings When zoom is in full out position zoom setting is at the mechanical minimum setting which is 1 0 for newer camera assemblies and 0 7 for older camera assemblies Zoom or position mechanisms should not bind Aiming the Camera 1 2 PN 42370036 Optimize the prism mounted on the scatter sensor a Loosen the securing setscrew b Rotate the adjusting lever to center the stream image in the strobe Optimize the periscope a Loosen the radiator hose clamp older units or the setscrews securing the periscope to the base newer units b Rotate the periscope 4 SERVICE AND REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE 4 1 10 3 Repeat steps 1 and 2 as needed until The stream is centered in the illuminated strobe area The strobe illumination is centered on monitor The stream and strobe illumination remain centered at all zoom settings Focusing the Camera The primary focus adjustment for the camera assembly is the upper internal mirror on the periscope Focus the camera according to the following procedure Loosen the securing setscrew at the upper internal mirror on the periscope 2 Move the mirror vertically until the best focus is achieved and tighten setscrew Note This adjustment affects the cameras position 3 Vary the zoom and ensure that the stream remains focused If the stream does not remain focused do step 4 4 Ifthe stream does not remain focused
408. mp Laser Alignment Time Space Separated Laser Alignment l Closethe Argon laser output shutter 2 Setthe UV laser power up or down to create a distinguishable difference in amplitude between the HeNe and UV laser resultant signals 3 Install the adjustable beam translator between the HeNe laser and its adjustable dichroic leaving space between the translator and the optical rail to mount a single laser target 4 Verify the following a The translator knobs face away from the laser b The translator window is perpendicular to the optical bench 5 Adjust the translator height in post for the laser to penetrate as close to the center of the translator window as possible 6 Refer to the multi hole target and set the alignment mode to Top 1 Move the pin on the bottom of the target to line up with desired target hole and lock pin down with the set screw 7 Install the middle target in the space you left between the beam translator and the HeNe dichroic ensuring the target is square with the table top 8 Rotate the translator window so the top of the Hehe beam passes through the middle of the top target hole 9 Remove the HeNe laser target PN 4237003 4 1 7 SERVICE AND REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE 10 11 12 13 14 15 16 1T 18 19 20 21 22 23 Adjust between the peak of the HeNe laser generated signal and the peak of the UV laser generated signal by usin
409. mping diode that prevents the output from going below approximately 300 mV The net result is that any signals smaller than 0 001 V will be put in channel 0 The log amp equation is Output Voltage n x log 9 1000 x Input Voltage Figure 2 6 3 is another graph of the log amp response The graph is now plotted on log lin axes The illustration shows the four decades 10 1 ratios covered by the log amp Figure 2 6 3 Log Amp Response Log Lin Axes 0 1 VOLTS IN 0 01 0 001 7003032A VOLTS OUT PN 4237003C 2 6 3 INSTRUMENT DESCRIPTION ACQUISITION SUBSYSTEM 2 6 4 The circuit is based on the non linear portion of a diode s V I curve The circuitry includes local voltage regulators and temperature compensation for stability These circuits are adjusted at the factory with specially designed test fixtures You cannot adjust the log amps in the field In Figure 2 6 4 operation of the circuit is based on two principles The gain is RI R2 RI e Resistor R2 is fixed but is the resistance of the diode Circuit elements not shown in the illustration ensure that the diode is biased to operate in the non linear portion of its V I curve Based on this for small input pulses the diode has a low resistance and the circuit gain is high As the input level increases the diodes resistance increases and the circuit gain drops The actual circuit is considerably more complex to ensure that the diode is correctly biased
410. n jet in air Now sort the 100 beads using delays 28 29 30 or whatever meets your criteria We use 28 29 and 30 here only because of our example in step 8 Check to see that most of the beads are now in 29 by looking at the drops on either side of 29 If there are less than 10 beads in drop 28 or drop 30 then there is gt 90 in drop 29 Go to step 13 If there 10 or more beads in drop 28 or drop 30 adjust the flow cell again either up or down depending on which direction the beads went Continue adjusting the flow cell until most or all of the beads are in the droplet of choice 29096 For example if you raised the flow cell too much the beads will fall in droplet 28 If you did not raise it enough some beads will fall in drop 30 If the flow cell was turned the wrong way then most of the beads will be in drop 30 Reduce the number sorted to 20 Do a sort matrix on the phase of that droplet 0 0 0 9 on the delay Note Several of the phases should have 20 in them Choose one phase and ensure that the side streams are good Verify the delay and phase by sorting several drops of 20 particles onto a slide and counting each For future use with this flow cell match the camera delay with the actual delay by using the flow cell adjustment on the Camera screen Note The instrument remembers this value until the software is reloaded at which point the value will have to be adjusted for that particular flow cell
411. n laser Refer to the appropriate figures for the laser being installed a HeCd Laser Figure 3 7 4 HeCd Laser Head Mounting Figure 3 7 5 HeCd Power Supply Connections Figure 3 7 6 HeCd Optical Arrangement Figure 3 7 7 HeCd Laser Configuration b Water cooled Laser Figure 3 7 8 1305 Mounting Figure 3 7 9 Water Cooled Optical Arrangement Figure 3 7 10 Water Cooled Laser Arrangement 3 7 6 PN 4237003 OPTIONAL LASER INSTALLATION INSTALLATION PROCEDURES 3 Figure 3 7 4 HeCd Laser Head Mounting 9 HeCd LAGER SCREW e eo 9 009 FRONT PLATE E WASHER MOUNTING BACK PLATE BOTTOM COVER OF PLATE REAR PLATE 7 PLATE SUPPORT FRONT OF INSTRUMENT 9 0 74 MOUNTING PLATE p d 7003212A Figure 3 7 5 Power Supply Connections HeCd POWER SUPPLY LINE CORD 7003213A PN 4237003C MOUNTING PLATE INTERLOCK CABLE 3 7 7 INSTALLATION PROCEDURES OPTIONAL LASER INSTALLATION Figure 3 7 6 HeCd Optical Arrangement 488 MIRROR BEAM REDUCER EXPANDER FLUORESCENCE HENE LASER ARGON LASER BEAM WATER COOLED COLLECTION OPTICS LASER 2 LASER 1 EXPANDER ARGON LASER o BEAM SHAPER 7003163A FLOW CELL LASER BEAM S 633 MIRROR BEAM STEERING BEAM TRANSLATOR ASSEMBLY Figure 3 7 7 HeCd Laser Configuration
412. nce the beam diameter is constant for a particular experiment the system can subtract this time from each width pulse by the Beam Width Normalization circuit this leaves a pulse that only represents the particle The card converts the pulse width to a voltage level that represents the length of the pulse This allows the system to acquire the level as a parameter The time to voltage circuit provides four scales to cover the range of possible pulse widths and particle sizes PPU Function The PPU function Figure 2 8 11 of the Pulse Pileup Det TOF card monitors the pulse for changes in slope polarity A single cell gives rise to one slope polarity change which is at the peak when the slope changes from positive to negative With a doublet the slope changes polarity at the peak of the first cell at the valley between the two cells and at the peak of the second cell When the PPU detects more than one slope change inside an event as determined by discriminator crossings the PPU generates an additional discriminator satisfied signal for the event The sorting electronics then use this information to abort a sort pulse if Abort is ON Note The PPU parameter also generates a positive detect output if the PPU parameter if that parameter is greater than 9 8 V or channel 1 000 PN 4237003C 2 8 17 INSTRUMENT DESCRIPTION SORT SUBSYSTEM 2 8 18 Figure 2 8 11 PPU Function of Pulse Pileup Det TOF Card CELL DETECTION WITHOUT PPU
413. ndirectly from reflective surfaces such as a mirror or shiny metal surface including tools Avoid direct exposure to the beam Do not view directly or with optical instruments except for special service instruments as directed in the service manual Wear latex surgical gloves to reduce the amount of UV in the event you accidentally place your hands in front of the beam 16 Open the laser output shutter on the UV laser 3 12 3 INSTALLATION PROCEDURES BEAM TRANSLATOR UPGRADE 3 12 4 1T 18 19 20 21 22 23 Align the UV laser a Use the 90 degree full mirror to align the UV laser through the two smaller vertical optical plate mounted targets b Use a business card or fluorescent day glow paper to detect the UV laser position Close the laser output shutter on the UV laser Place the screw on targets input and output lenses on the beam reducer Note Older style beam reducers can be used but there are no built in targets Install the beam reducer on the optical bench as close as possible to the output of the UV laser Note Allow enough room to remove the screw on targets from the beam reducer when you are finished Open the UV laser output shutter Align the X Y Z axis of the beam reducer through a The two optical bench mounted targets b The two vertical optical plate mounted targets After the beam reducer is properly aligned remove the screw on targets from the beam reduc
414. ne by the customer Tools Supplies Needed Version 4 0 Software Kit PN 6912816 which includes gt Disk 1 PN 6414976 8 gt Disk 2 PN 6414977 6 gt Field Correction Notice PN 4236307 3 PN 4237003C 3 18 1 INSTALLATION PROCEDURES SOFTWARE VERSION 4 0 INSTALLATION Procedure l Insert the Program Distribution Disk 1 of 2 disk in the 3 5 in floppy drive 2 Atthe DOS command a B and press Enter b Type INSTALL and press Enter 3 Follow the instructions that appear on the screen Remove the disk from the floppy drive 5 Reboot the computer by pressing ci Alt Delete Note if you exit from the Elite program type CYTOMETER and press Enter Refer to the README DOC file for additional instructions Transfer the new control file to the Cytometer by selecting Acquisition rr Start at the Workstation Do Heading 5 1 VERIFICATION INSPECTION PROCEDURE Provide the customer with a completed Field Correction Notice 10 Verify that the customer s manual is updated Note Updated manuals are also included in PN 6913228 6 3 18 2 4237003C INSTALLATION PROCEDURES SOFTWARE VERSION 4 01 INSTALLATION 3 19 SOFTWARE VERSION 4 01 INSTALLATION ATTENTION This procedure is mandatory Purpose Do this procedure to install Elite Software Version 4 01 in all instruments currently with version 4 0 As noted in the README DOC file version 4 01 contains several enhancements including change in
415. nel to provide access to Motor Controller card cage Open the panel on front of the Cytometer left of the sample delivery door Remove the optical filters from optical block Remove the optical block cover PN 4237003C PN 42370036 14 15 16 17 18 19 20 INSTALLATION PROCEDURES AUTOCLONE SORTING OPTION INSTALLATION If the lip on the laser interlock assembly where the assembly fastens to optical table near the flow cell area does not face the front of the Cytometer and if the bolt heads are not exposed a Remove the laser interlock assembly b Remove the left side panel from laser interlock assembly c Transfer components from laser interlock assembly panel just removed to new laser interlock assembly panel d Place new foam on new panel using old panel for reference e Install laser interlock assembly on optical table Remove the Motor Controller card from the lower slot Install the connector mounting block over 44 pin card edge connector of the cable assembly a Make sure the stubs with the threads are towards the side of the wires b Work the connector into the Cytometer from where the sample collection drawer was to the rear of the Motor Controller card cage Place the 44 pin connector into the empty slot above the Motor Controller card slot and verify that the letters side is up Fasten the screw to the threads in the connector mounting block a the screw to a Phillips head sc
416. ng 2 6 1 Subtraction Color Compensation 2 6 1 Log Amplification 2 6 2 Integration 2 6 4 Special Functions 2 6 5 Switchable Amplifiers Bandwidth 2 6 5 Gated Amp Concept 2 6 6 Signal Flow for Each Configuration 2 6 8 4 PMTs Nonswitchable Amps 2 6 9 4PMTs Switchable Amps with Peak Scatter Function 2 6 11 5PMTs Nonswitchable Amps No Gated Amp 2 6 13 5PMTs Switchable Amps No Gated Amp 2 6 15 Gated Amp 2 6 17 OPTICAL ELEMENTS 2 7 1 Overview 2 7 1 Light 2 7 1 Definition 2 7 1 Media Used for Travel 2 7 1 Snell s Law 2 7 2 Lenses 2 7 2 Positive Lens 2 7 2 Negative Lens 2 7 3 Equations 2 7 3 Optical Components Used in the Elite 2 7 4 Beam Shapers 2 7 5 Beam Expander Reducer 2 7 7 Parallel Plate 2 7 7 Light Sources 2 7 8 High Sensitivity Flow Cell Tips 2 7 8 SORT SUBSYSTEM 2 8 1 Function 2 8 1 Overview 2 8 1 Independent Operation 2 8 1 Sort Cycle 2 8 1 Circuit Cards 2 8 1 Prism and Sort Window Test Card 2 8 4 Bitmap and Sort Decision Card 2 8 5 Interface and Scaler Card 2 8 7 Sort Delay Card 2 8 8 vii viii 2 0 2 10 2 11 2 12 Sort Oscillator Card 2 8 10 Function 2 8 11 Sort Output Card 2 8 13 Pulse Pileup Det TOF Card 2 8 15 TOF Function 2 8 16 PPU Function 2 8 17 Transistor Bracket Card 2 8 19 Power Supplies 2 8 19 PNEUMATICS SUBSYSTEM 2 9 1 General 2 9 1 Solenoid Valves 2 9 1 Circuit Cards 2 9 4 Pneumatic Interface Card 2 9 4
417. no beam translation Adjust the positioner knobs to target the UV beam Note Further adjustments depend on the customer s specific operating needs the beam translators can be used to shift the beams to be coaxial or to place either beam in the upper position As an initial test align the beams to be coaxial PN 4237003C INSTALLATION PROCEDURES ENTERPRISE LASER UPGRADE Figure 3 14 1 Enterprise Laser Mounting Location HOLE 12 3 OLE 1 1 ENTERPRISE LASER EE Ter icm er 9 4 HOLE 7 4 0 007050 100 0 ken Jan ak PAN REN Vel 7003165 Figure 3 14 2 Beam Separator Location ND FILTER VARIABLE ND FILTER USED TO ADJUST 488 POWER LOCK CAN ALSO BE USED TO ADJUDT UV POWER 488 nm TRANSLATOR UV TRANSLATOR a 1 Ps M UV BEAM POINTING VERTICAL Uv BEAM UV FOCUS POINTING BEAM SIZE HORIZONAL 7003164A 11 Install the short targets in the flow cell area 12 Remove the tall targets and install the optical rail elements 13 Remove the flow cell tip and beam shaping optics 14 Begin with the front HeNe laser use the optical rail beam positioners to target each beam Be sure all adjustments are secure when done 15 Reinstall the flow cell beam shaping optics and scatter sensor PN 4237003C 3 14 3 3 INSTALLAT
418. no gated amp Gated Amp circuit cards in 2 12 1 HeCd laser 3 7 8 non Gated Amp circuit cards in 2 11 1 connections See cable connections converter DAC See DAC cooling coil See Compressor module cooling fans See fans CPU 835 Intel card jumpers 4 10 1 A 2 2 CPU IBC 86C card jumpers 4 10 1 A 2 2 CRT dual pathways display source 2 5 4 crystal drive adjustment procedure 4 2 2 test procedure 4 2 2 verifying sort pulses are synchronous with 4 2 4 customer training required before instrument installation 3 1 1 CW See Autoclone Sorting Option 2 2 3 Cytometer circuit cards functions of 2 5 6 decontamination procedure 4 11 3 installation procedure 3 2 1 verifying operation of 3 3 1 Cytometer Computer subsystem description 2 5 1 Cytometer Control subsystem circuit cards 2 5 6 function 2 5 6 Cytometer CPU card function 3 22 1 4 13 1 installation 3 22 1 4 13 1 obsoletes 256K Memory card 3 22 1 4 13 1 D DAC defined 2 9 4 function of 2 9 4 2 13 4 reference voltage measurement of 2 9 4 DAC card function of 2 5 8 Data Acquisition card cage circuit card jumpers 4 10 3 Data Lister Out card digital values reading 2 13 1 6 INDEX jumpers 4 10 3 A 2 3 Data Taker Interface card function 2 5 6 jumpers 4 10 1 A 2 2 decontaminating the system 4 11 3 decontamination documentation Biohazards Safety Guide 1 2 1 Classifications of Etiological Agents on the Basis of Hazards 1 2 1 source mater
419. ns Elite and Elite Analyzer See Table 8 2 10 REAR OF UNIT 7003089A Table 8 2 10 Elite and Elite Analyzer Interconnections Detail of Parts See Figure 8 2 10 Part Number Description Item Part Number Description 1 6028210 Backflow cable 8 6028022 Bitmap Memory card cable 2 6028374 Analyzer card W52 cable 9 6028023 Scope signal cable 3 6028020 Sensor interface cable 10 6028038 Data Acquisition card cable 4 6028018 Sort transistor cable 11 6028024 CPU Pneumatics card cable 5 6028040 36 V W19 cable 12 6858355 Connector panel Elite Analyzer 6 6856953 Cable harness W12 13 6028070 BNC W31 cable D 6028025 Data Lister Out cable 14 6028021 Serial communication cable 8 2 10 PN 4237003C PARTS LISTS 8 ILLUSTRATED PARTS Figure 8 2 11 Multibus Card Cage All Configurations See Table 8 2 11 1 7003120A H Table 8 2 11 Multibus Card Cage All Configurations Detail of Parts See Figure 8 2 11 Item Part Number Description 1 6704933 Computer backplane Cytometer 2 6706571 Computer IBC 86C card with on board memory Note If you are replacing an old Intel CPU 8086 card 2016414 remove the 256K Memory card PN 2016494 also Once the new computer card is installed the Cytometer Software must be downloaded from the Workstation 3 6702664 Serial 1 0 card
420. nside Rear View Detail of Parts See Figure 8 2 13 8 2 13 Cytometer Right Rear Door Inside View Detail of Parts See Figure 8 2 14 8 2 14 Cytometer Left Rear Door Inside View Detail of Parts See Figure 8 2 15 8 2 15 Cytometer Left Front Corner Inside View See Figure 8 2 16 8 2 16 Analyzer Sample Drawer Detail of Parts See Figure 8 2 17 8 2 17 Analyzer Top Inside View Detail of Parts See Figure 8 2 18 8 2 18 Elite Flow Cell Area Detail of Parts See Figure 8 2 19 8 2 19 Elite Analyzer Flow Cell Area Detail of Parts See Figure 8 2 20 8 2 20 Deflection Body Detail of Parts See Figure 8 2 21 8 2 21 Photomultiplier Tubes PMTs Detail of Parts See Figure 8 2 22 8 2 22 Autoclone Sorting Option Mechanism Detail of Parts See Figure 8 2 23 8 2 23 Autoclone Sorting Option Beak Waste Catcher Detail of Parts See Figure 8 2 24 8 2 24 PN 4237003C 8 8 2 25 Autoclone Sorting Option Location and Connections Detail of Parts See Figure 8 2 25 8 2 25 8 2 26 Autoclone Sorting Option Accessories Detail of Parts See Figure 8 2 26 8 2 26 8 2 27 Transformer Drawer Assembly Detail of Parts See Figure 8 2 27 8 2 27 8 2 28 Computer Box Assembly Detail of Parts See Figure 8 2 28 8 2 28 8 2 29 Beam Splitter Dichroic Detail of Parts See Figure 8 2 29 8 2 29 8 2 30 Beam Shaper Detail of Parts See Figure 8 2 30 8 2 30 8 2 31 Elite ESP Flow Cell Stage Detail of Parts See Figure 8 2
421. nside View Detail of Parts See Figure 8 2 15 Item Part Number Description 1 2603065 Fan blower 112 CFM 115 Vac 2 6705493 Deflection power supply 3000 V PN 4237003C 8 2 15 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 16 Cytometer Left Front Corner Inside View See Table 8 2 16 1 FRONT VIEW OF LEFT SIDE OF PEDESTAL 7003014A WITH COVER REMOVED LEFT FRONT RIGHT CORNER OF CYTOMETER Table 8 2 16 Cytometer Left Front Corner Inside View See Figure 8 2 16 Item Part Number Description 1 2802006 Obscuration bar jet in air 2 6857598 Jet in air ring 3 1019958 Screw machine 2 56 x 0 19 4 2807038 Setscrew 6 32 x 0 250 5 6857636 Interlock bypass adapter 6 6857637 Interlock relay bracket 8 2 16 4237003 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 17 Analyzer Sample Drawer See Table 8 2 17 CT ege 7003132 Table 8 2 17 Analyzer Sample Drawer Detail of Parts See Figure 8 2 17 Item Part Number Description 1 1022054 Splash shield 2 6858923 Sample cap 3 3210004 Sample tubing 0 010 i d insertion location 4 1018859 Pinch valve cover 5 1020913 Block sample station 6 6858907 Door 7 1019224 Pickup probe 8 1017916 Base 9 6232205 Fitting hose barbed elbow 0 125 i d to 10 32 thread PN 4237003C 8 2 17 8 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 18 Analyzer Top Inside View See Table 8 2 18 EISE oe 7003144A 17
422. nsity staining sense air cooled umsec Tissue 140 um sort 15 mW 15 x 80 um 15 umsec Low gt 20 Medium culture hybridoma sense air cooled cells 20 40 um diameter Enrichment 76 Fast 200 mW 15 x 33 um 2 5 umsec High 0 5 4 0 High high speed sorts Quartz water cooled umsec DNA stained with PI 100 um Sort 15 mW 8 x 80 um 5 umsec Low 4 0 10 0 Low Sense air cooled umsec 2 1 6 42370036 INSTRUMENT DESCRIPTION OPTICAL ELEMENTS Table 2 7 1 Typical Applications of Beam Shapers Continued Beam Shaper DISCSAT Band PPU Data Sorting Application Flow Cell Laser Power Spot Size EXT Width Sensitivity Rate Chromosomes 76 or 100 50 457 6x100 um 5 umsec Low 4 10 Low um Sort nm Sense 50 mW UV Stem cells 100 um Sort 15 mW 15 x 80 um 5 Low 4 10 High Bone marrow Sense air cooled msec Beam Expander Reducer This optical system of the beam expander reducer Figure 2 7 7 consists of a positive and negative lens The lenses are situated so that their focal points overlap Figure 2 7 7 Beam Expander Reducer FOCAL LENGTH OF NEGATIVE LENS lt FOCAL LENGTH OF POSITIVE LENS 7003177A As a laser beam which consists of parallel rays enters the negative lens it is refracted to diverge from the focal point of the negative lens This results in a parallel laser beam with an increased diameter beam ente
423. nt stage and one to the Y movement stage Each card has two optical sensors and two mechanical switches The optical sensors detect holes punched in a fixed metal strip indicating the normal travel limits of the stage mechanism The mechanical micro switches are present only to detect if accidental overtravel has occurred and to advise the operator to calibrate the system 2 2 3 2 INSTRUMENT DESCRIPTION AUTOCLONE SORTING OPTION 2 2 4 Software ATTENTION Autoclone Sorting Option software resides in both the Workstation and the Cytometer The Workstation software obtains the necessary information from the user the type of tray which wells to sort into number of cells to put into each well and the sort criteria for each well The Workstation transfers the information to the Cytometer through the serial communication path The Cytometer software physically operates the Cytometer mechanism to operate the motors and interpret information from the optical and mechanical sensors It also supports the Autoclone Sorting Option Control and the Adjust and Retract screens on the Cytometer Through these screens you can initiate calibration and adjustment routines Calibration The calibration routine operates each stepper motor in turn to find the limits of travel using the optical sensors and to count the number of steps between the two limits With this information the Cytometer computer knows where the mechanism is and can calculate
424. nterface Card A 1 2 Power Supplies A 1 2 Optics A 1 2 Beam Expander A 1 2 Signal Adjustments A 1 2 Beam Translator Upgrade A 1 2 Gated Amp Laser Alignment Time Space Separated Laser Alignment A 1 3 2 JUMPERS AND SWITCH SETTINGS A 2 1 Jumpers for Circuit Cards Used During Installation and Upgrade Procedures A 2 1 3 PMT Sub SW R2 Card A 2 1 HV DAC Card A 2 1 HV DAC Control Card A 2 1 HV DAC Control Card Gated Amp 5 PMT option A 2 1 Lister AT Card on PC Model 486 A 2 1 Mux and Scope Card A 2 1 Sensor Interface Card A 2 1 Jumpers for Cards in Card Cages A 2 2 Multibus Card Cage A 2 2 Non Gated Amp Card Cage A 2 2 Gated Amp Card Cage A 2 3 Data Acquisition Card Cage A 2 3 Jumpers for Line Voltages A 2 4 OPTICAL SETTINGS A 3 1 Collection Optics Upgrade A 3 1 Sort Waveform Verification A 3 1 Crystal Drive Test Procedure A 3 1 Crystal Drive Adjustment Procedure A 3 1 Sort Pulse Amplitude Test A 3 1 Power Supplies A 3 2 B FILE PATHS B 1 1 LOCATION OF SOFTWARE FILES B 1 1 PN 4237003C 9 i 9 1 ILLUSTRATIONS B 1 1 TABLES 1 1 2 1 2 2 2 3 2 4 2 5 3 1 File Paths B 1 1 Pneumatic Interface Card Adjustments A 1 2 Multibus Card Cage Jumpers and Switch Settings A 2 2 Non Gated Amp Card Cage Jumpers A 2 2 Gated Amp Card Cage Jumpers A 2 3 Data Acquisition Card Cage Jumpers A 2 3 Line Voltage Jumpers A 2 4 Power Supply Adj
425. nterface card Connect the external pressure gauge with a Y fitting to the sheath pressure line at the sheath pressure pinch valve center fitting 7 At the Valves screen a Adjust Sheath SET pressure until the external gauge reads 15 psi b Adjust the Range control until the DVM reads 10 0 V Note The maximum pressure the Pneumatic Interface card accepts is 15 psi If the external gauge reads less than 15 psi when 15 psi is entered in this step adjust R51 to obtain 15 psi c Adjust Sample SET pressure until the external gauge reads 3 psi d Adjust the Zero control until DVM reads 1 0 V Repeat steps 7 until no further adjustment is needed Readjust the Pneumatic Interface card 4237003 4 6 1 4 SERVICE AND REPAIR PROCEDURES ELECTRONIC PRESSURE REGULATOR ADJUSTMENT 4 6 2 PN 4237003 SERVICE AND REPAIR PROCEDURES PRESSURE SWITCH ADJUSTMENT 4 7 PRESSURE SWITCH ADJUSTMENT Purpose Use this procedure to adjust the system pressure switch Tools Supplies Needed DVM Procedure 1 2 PN 4237003C Turn the unit off Connect the DVM to the two terminals on the system pressure switch while leaving the system wires attached Set the DVM for DC V Turn the unit ON Observe the DVM Note The reading is 4 to 5 V immediately after you turn the system on then the voltage suddenly drops to 0 V after compressor pressure has built up At the Valve Screen turn main pressure OFE Adjust the
426. o bimorph plate optical reinstallation of 3 15 1 PMI procedure 5 2 1 PMT Gated Amp card block diagram 2 12 5 function 2 12 4 inputs signal 2 12 4 jumpers 4 10 3 A 2 3 PMT Gated Amp R card jumpers 3 9 4 Pneumatic Interface card adjustment procedure 4 5 1 communication pathways from Multibus CPU card 2 9 4 function 2 9 4 jumpers 4 5 2 jumpers for troubleshooting pressure control problems 7 4 2 7 4 3 potentiometer adjustments for use with low bleed regulator 4 5 1 potentiometer adjustments for use with non low bleed regulator 4 5 4 pressure regulators use of 2 9 4 R31 adjustment 10 00 0 01 Vdc 4 5 1 A 1 2 R34 adjustment 10 00 0 01 Vdc 4 5 1 A 1 2 R51 adjustment 8 00 0 01 Vdc 4 5 1 A 1 2 R54 adjustment 10 00 0 01 Vdc 4 5 1 A 1 2 R81 adjustment 10 00 0 01 Vdc 4 5 1 A 1 2 regulation process control of 2 9 5 solenoid valve operation affect on 2 9 4 test points potentiometer adjustments 4 5 1 pneumatics circuit cards list of 2 9 4 components illustration of 7 3 1 Compressor See Compressor module electronic pressure regulator adjustment 4 6 1 gauge pressure defined 4 5 1 low bleed regulator adjustment 4 5 1 non low bleed regulator adjustment 4 5 4 read pressure defined 4 5 1 sample pressure system new type See sample pressure system new type sample pressure system old type See sample pressure system old type sample regulator adjustment 4 5 3 12 INDEX set pressure
427. o DOS directory Run SCANDISK on C System Not Serviced Within Past 30 Days 1 SB 907 mU Sr MI Inspect and adjust compressor module as needed for correct pressures Verify that the Compressor module is freestanding on the floor and does not touch the instrument Perform Powerup Valves at the Cytometer Set Sheath and Sample pressures to 13 psi At Valves screen ensure that SET and READ pressures agree within 0 05 psi Note If pressures do not agree do Pneumatic Interface card calibration procedure Power up all lasers Ensure blowers are operating Verify operation of pneumatic laser shutter Verify operation of mechanical laser shutter Verify operation of sample vial detect Operational Test Cytometer Electronics Optics or Fluidics Do this procedure if repairs were made affecting Acquisition CV or Analysis subsystem 1 PN 42370036 Create a protocol to acquire fluorospheres as follows or use saved protocol suitable for acquiring fluorospheres e Histogram 1 FALS 9015 stop at 5000 Histogram 2 FALS e Histogram 3 9015 MAINTENANCE PROCEDURES VERIFICATION INSPECTION PROCEDURE Histogram 4 Log GFL e Histogram 5 GFL e Histogram 6 Peak GFL FALS discriminator 100 all others OFF e All gains 10 All HV 400 Select Acquire and ensure values transfer correctly to Cytometer Adjust settings as necessary to obtain good histograms Analyze for CV value Ensure that the results a
428. o deu c OUT x E EE bog 1 1 o gomme 3 m PENNE HER S E Le a T H Set by 1 0 These commands originate Gated pulse from Ji command from at Gated Amp control where Gated Amp Control Card P1 SCAT AUX CPU they are decoded from BACKPLANE TRIGGER commands J6 J10 NM Les as pH TN 1 42 18 AS DC RESTORE FALS OUT J3 J19 1 DC RESTORE 1 IN OUT 7003045A 2 12 3 INSTRUMENT DESCRIPTION CARD DESCRIPTION GATED AMPLIFIER CONFIGURATION PMT Gated Amp Card Function The PMT Gated Amp card Figure 2 12 3 Selects one of the four signal inputs to be the trigger source and routes it to the Gated Amp Control card Selects two signals to be AUX1 and AUX2 from the four signal inputs and sends the signals to the Scat Aux Gated Amp card Sends the four input signals to the delay cards For each channel the PMT Gated Amp card can use the returned delayed signal or the undelayed signal e Gates by switching the signal outputs between the signal inputs or ground The gate pulse from the Gated Amp Control card controls the gating Inputs The four signal inputs for this card are e PMT2 e PMT3 e PMT4 e PMT5 212 4 4237003 INSTRUMENT DESCRIPTION 2 CARD DESCRIPTION GATED AMPLIFIER CONFIGURATION Figure 2 12 3 Block Di
429. o the Acquisition electronics where the PMT2 was removed 9 At the Cytometer s Scope screen a Set PMT2 Peak on the lower trace b Set PMT2 Int on the upper trace C Set the time base to 20 microseconds division 10 At the Main screen adjust PMT 2 Gains so the pulses are between 2 to 3 divisions high Acquire and Setup Sort l Atthe Workstation select Acquisition Parameter 2 Erase all parameters 3 Atthe Create mode select PMT2 Peak and PMT2 Note The list of parameters in the right column should appear as PMT2 Peak PMT2 If PMT2 Peak is not the first parameter repeat this step If PMT2 Peak is the first parameter go to step 4 4 At the Cytosettings screen a Setthe PMT2 Peak Discriminator to 100 b Setother channels to OFE PN 4237003C PN 4237003C Du 9o cub OON 11 12 13 14 15 16 1T 18 SERVICE AND REPAIR PROCEDURES SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE At the Protocol screen create two single parameter histograms to acquire PMT2 and PMT2 Peak Press At the Cytometer screen put the PMT2 Peak histogram at mid scale Restart build a histogram then Stop At the Workstations Sort screen create a sort region around the PMT2 Peak distribution and assign to Sort Right Start acquisition and send sort settings to the Cytometer At the Cytometer a Set Sort Stop counters to off and turn Sort Right Enable on and start sort right b Set Drive Frequency to 32 kHz
430. occur downstream sufficiently focused and clean drops Satellite drops never Bubble or clog in flow cell Press CLEAR three times then press disappear DEBUBBLE Drops are Drops not stable for Instabilities with fluidics 1 Change the sheath pressure slightly up or unstable after a period of time more than 3 or 4 hours down x1 0 to 2 0 psi 2 Complete sort setup beginning with Drive Frequency Sort Settingsunder Heading 4 2 More than a 10 15 change in the crystal drive is needed to achieve the droplet breakoff attained earlier that day Bimorph is unstable Replace the bimorph and verify it is tightly secured PN 4237003C 7 5 7 7 TROUBLESHOOTING TROUBLESHOOTING THE SORT SUBSYSTEM Figure 7 5 2 Satellite Droplets CURSOR PLACEMENT FOR MONITORING LAST ATTACHED DROPLET o aa THIN FILAMENT BETWEEN LAST DOWNSTREAM ATTACHED DROP AND STREAM SATELLITE DROPLETS RECOMBINING USE THE CRYSTAL DRIVE ADJUSTMENT 10 TO KEEP THIS THIN 7256008 7 5 8 4237003 TROUBLESHOOTING RESETTING CYTOMETER SOFTWARE LOCKUPS 7 6 RESETTING CYTOMETER SOFTWARE LOCKUPS ATTENTION These reset techniques only apply to those Cytometers containing an External Memory card The older 256K Memory card does not respond in the same manner Purpose Reset the CPU s memory anytime the Cytometer is locked up not responding properly If after resetting the Cytometer CPU the Cytomet
431. ograms kHz kilohertz kV kilovolt kW kilowatt kPa kilo Pascal L l long L left length liter lock laser light amplification by stimulated emission of radiation Ib pound LCD liquid crystal diode LED light emitting diode Li lithium Im lumen LP long pass LT digiscope lower trace BNC connector low volume LWR BM lower beam m meter mA milli amperes max maximum MB megabyte MCB Motor Control board card MEM memory MF manifold 4237003C min minute misc miscellaneous MF manifold MHz mega Hertz min minimum minute mL milliliter mm millimeter msec microsecond mV millivolts mW milliwatt N A not applicable NB ND neutral density nm nanometer 0 o d outside diameter op amp operational amplifier 02 ounces P param parameter PC personal computer PCB printed circuit board card pf PGEN Pulse Generator and Clock R card PK SCAT Peak Scatter Mux SW R card PM pump PMI preventive maintenance PMT photomultiplier tube PMT GA PMT Gated Amp P part number NEU pneumatics pot potentiometer ppm parts per million PPU pulse pileup preamp preamplifier PN 4237003C ABBREVIATIONS PROM programmable read only memory PS power supply PSH peak sense and hold psi pounds per square inch PVC polyvinylchloride PWR power 0 QPSH amp H q
432. olic Mirror on Tip Optional l Atthe Cytometer a Select Options b Change Align LED to Continuous c Select Main 2 Insert a mirror into the first filter slot on the right The LED backlights the pinhole and provides an image marking the center of the assembly 3 Close the laser shutter Center the LED image in the parabolic mirror See Figure 4 12 7 Adjust the flow cell vertical knob as needed Adjust the flow cell horizontal knob as needed 4 12 4 4237003C SERVICE AND REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE Figure 4 12 7 Parabolic Mirror and LED p ee O WC PARABOLIC MIRRORS B 7003198 LED 5 Turn off the LED Remove the mirror from the first slot on the right Do Laser Beam Alignment Laser Beam Alignment 1 Press SHEATH Open the laser shutter See Figure 4 12 8 for location 2 Figure 4 12 8 Laser Shutter Location FLOW CELL LASER BEAM INTERLOCK VERTICAL VERTICAL DEFEAT BAR ADJUSTMENT ADJUSTMENT FLOW CELL Z AXIS ADJUSTMENT FLOW CELL HORIZONTAL ADJUSTMENT LAGER HORIZONTAL TA 6 ADJUSTMENT e LAGER SHUTTER 7003196A BEAM SHAPER PN 4237003C 4 4 12 5 SERVICE AND REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE 4 12 6 3 Center the laser beam vertically on the forward scatter detector mask by adjusting the laser beam vertical kno
433. on Part Number Description 1 2016414 Intel CPU 8086 card 20 16704188 Sort Oscillator card 2 6702664 Serial 1 0 card 21 6704201 Sort Output card 3 6703761 Data Taker Interface card 22 6702608 Sort Transistor card 4 7000681 External Memory card 23 6704555 Sensor Interface card 5 6704089 Dual CRT Controller card 24 6704164 Mux and Scope Interface card 6 6703772 Digiscope card 25 6705252 Scat CV SW R1 card 7 6704463 Dual Laser Control card 26 6705369 Peak Scatter Mux SW card 8 6704660 Camera Interface card 27 6705246 3 PMT Sub SW 1 card 9 6704907 Pulse Generator and Clock card 28 6704009 Scat Aux Gated Amp card 10 6704959 Pulse Pileup Det TOF card 29 6704904 Quad 20 40 60 Microsecond Delay 1 card 11 6704164 Mux and Scope Interface card 30 16704241 Quad 7 Microsecond Delay 1 card 12 6704169 Quad PSH 2 card 31 6704241 Quad 7 Microsecond Delay 2 card 13 6704169 Quad PSH 1 card 32 6704904 Quad 20 40 60 Microsecond Delay 2 card 14 6705400 Peak ADC PSH Control card 33 6704008 PMT Gated Amp card 15 16704142 Data Lister Out card 34 6705246 3 PMT Sub SW 1 card 16 6704185 Prism and Sort Window card 35 6705246 3 PMT Sub SW 2 card 17 6704147 Interface and Scaler card 36 6705249 Dual FL SW R card 18 6705681 Analyzer Interface card 37 N A Spare slot 19 6704198 Sort Delay card 38 6705327 Gated Amp Control R3 card PN 4237003C 8 2 9 8 PARTS LISTS ILLUSTRATED PARTS Figure 8 2 10 Cable Interconnectio
434. on 4 2 4 Drive Frequency Sort Settings 4 2 4 Phase Adjustment 4 2 5 Delay Calculation 4 2 7 Monitoring the Sort 4 2 8 Sort Coincidence Verification 4 2 9 Tools Supplies Needed 4 2 10 Setup Pulse Generator 4 2 10 Acquire and Setup Sort 4 2 10 Pulse Pileup Verification 4 2 11 Verification of Sort Pulse Extension 4 2 12 Sort Pulse Truncation Test 4 2 12 Extended Abort Test 4 2 13 Pulse Generator Card Adjustment 4 2 14 LINE VOLTAGE SELECTION 4 3 1 POWER SUPPLIES MEASUREMENT AND ADJUSTMENT 4 4 1 Purpose 4 4 1 Tools Supplies Needed 4 4 1 Procedure 4 4 2 xiii xiv 4 5 4 6 4 7 4 8 4 9 4 10 411 PNEUMATICS SYSTEM 4 5 1 Low Bleed Regulator Adjustment Procedure 4 5 1 Purpose 4 5 1 Tools Supplies Needed 4 5 1 Pneumatic Interface Card Adjustment 4 5 1 Low Bleed Regulator Adjustment 4 5 2 Non Low Bleed Regulator Adjustment Procedure 4 5 4 Purpose 4 5 4 Tools Supplies Needed 4 5 4 Procedure 4 5 4 ELECTRONIC PRESSURE REGULATOR ADJUSTMENT 4 6 1 Purpose 4 6 1 Tools Supplies Needed 4 6 1 Procedure 4 6 1 PRESSURE SWITCH ADJUSTMENT 4 7 1 Purpose 4 7 1 Tools Supplies Needed 4 7 1 Procedure 4 7 1 WORKSTATION AND SOFTWARE 4 8 1 PC Model 486 Setup Procedure 4 8 1 SOFTWARE INSTALLATION 4 9 1 Purpose 4 9 1 Tools Supplies Needed 4 9 1 Procedure 4 9 1 ELECTRONIC ADJUSTMENTS CABLE AND JUMPER CONFIGURATIONS 4 10 1 Purpose 4 10 1 Tools Supplies Ne
435. on air cooled and the HeNe lasers to be positioned also Figure 3 9 7 shows a standard laser configuration 3 9 22 4237003C INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE Figure 3 9 7 Standard Laser Configuration ECD PE APC 670 me SU S827 640 LP DICHROIC FILTER ARRANGEMENT STANDARD 7003018A Laser Alignment 1 PN 42370036 Install the front 457 nm laser using both targets with no optics The beam should go cleanly through both targets to ensure a beam parallel to the optical bench a Place a piece of paper or some non flammable material on the wall opposite the laser to establish a wall target b For Elite configured units once the laser is level lower the laser 1 16 inch by marking a spot below each target and checking them one at a time since the beam will no longer go through the target holes c For Analyzer configured units once the laser is level raise the laser 1 16 inch by marking a spot below each target and checking them one at a time since the beam will no longer go through the target holes Install the back UV laser using the targets Do not move the laser Measure the spots on the wall between both beams to ensure the distance apart is the same as the laser beam exit point on each laser Note Both lasers should be exactly parallel
436. one Procedure 1 Mount the tall targets in front of the HeNe laser 2 Remove the HeNe dichroic if present 3 Turn ON the HeNe laser 4 Adjust the HeNe mounting to align the beam through the tall targets Use the center holes on the three hole targets to align the beam b Adjust the HeNe by moving the laser feet for horizontal movement Adjust vertical movement by loosening the lower screw on the laser foot adjusting the upper screw to move the laser Retighten the lower screw 5 Remove the tall targets and install the short targets in place of the scatter sensor and next to the laser shutter Install the HeNe dichroic in front of the HeNe Adjust the dichroic to align the beam through the targets If the beam passes through the target closest to the laser but does not align with the target in the scatter sensor position the output end of the HeNe laser needs a small adjustment Work with the HeNe adjustment and the dichroic until the beam passes cleanly through both targets a Turn Hehe laser off b Turn Argon laser on 8 Align the Argon laser beam through the targets Adjust the Argon laser mounts to align the laser beam through the targets b Make vertical adjustments by loosening the upper nut on the mounting post and adjusting the lower nuts then re tightening the upper nuts c Make horizontal adjustments by loosening the screws securing the laser to the base and sliding the laser then retightening Note As a gener
437. ons to be sorted Direction left or right Droplet as a measure of time the reciprocal of the droplet frequency gives the period of one droplet At 32 KHz one droplet period is 1 32 KHz therefore 1 32 KHz 31 microseconds This allows you to relate the time difference between pulses as observed on the scope relative to how far apart they will be in the sort stream in droplets Followed by implies the second cell is close enough to the first cell to create a sorting problem 2 8 12 PN 4237003C INSTRUMENT DESCRIPTION SORT SUBSYSTEM Table 2 8 1 Coincident Condition Responses Continued Condition Response System Response Explanation Bad cell Abort with If Abort is ON and Complete Abort is not selected an abort occurs followed by a Complete only if the bad cell is in the same droplet as the good cell which good cell Abort not means no more than four quarter droplets away ON This means that Note It is possible for a bad cell to occur early enough in the first the bad cell droplet and not be detected by the system which means the system does not meet sorts it with the good cell when greater than a 1 drop sort envelope the criteria to is chosen sort in either direct Abort with If Abort is ON and Complete Abort is selected an abort occurs if the VR Complete droplet window that the bad cell would have generated and the Abort ON droplet window for the good cell overlap For example In a three drop
438. oportional to the digital pressure Full Scale is set by Volt Ref PN 4237003 2 9 11 INSTRUMENT DESCRIPTION PNEUMATICS SUBSYSTEM Compressor Mo The Compressor module Figure 2 9 6 contains a single compressor with two heads and provides the system with pressure and vacuum The module operates on 110 Vac 50 60 Hz A transformer converts for different voltages dule Figure 2 9 6 Block Diagram Compressor Module VACUUM CHOKE WHITE QUICK CONNECT AIR IN COMPRESSOR Y 30 PSI DUMP VACUUM SOLENOID MUFFLER SYSTEM PRESSURE RELIEF VALVE DUMP SOLENOID REGULATOR CLOSED PORT 0 60 PS SYSTEM PRESSURE BLUE SYSTEM PRESSURE Mounting brackets secure the Compressor module for transport However for normal operation the module must be set on the floor and cannot touch the instrument 29 12 7003078 INSTRUMENT DESCRIPTION CARD DESCRIPTION ALL CONFIGURATIONS 2 10 CARD DESCRIPTION ALL CONFIGURATIONS FL1 IN J3 FL2 IN J5 FL3 IN 3 PMT Sub SW R Card The 3 PMT Sub SW R card accepts three FL inputs Each system can have a percentage of one or both of the other inputs subtracted from it After subtraction the card outputs the FL1 and FL2 signals The card processes the FL3 signal to provide variable gain peak and integral signals and a log output See Figure 2 10 1 Figure 2 10 1 3 PMT Sub SW R Card Block
439. or air bubbles If necessary press DEBUBBLE and VACUUM as necessary to remove bubbles Display the Sheath and Sample Pressures on the Cytometer screen v 0 Verify that both sheath and sample pressures are 12 psi Note If the set and read pressures do not agree to 0 5 psi do the procedure under Heading 4 5 PNEUMATICS SYSTEM 10 Verify the sample vial detector is operating correctly Camera ATTENTION For Elite Analyzer installation skip this procedure l Install the deflection plate Figure 3 5 3 and push securely into position Figure 3 5 3 Deflection Plate 7003107A 2 Activate the strobe 3 Setthe left Cytometer screen to the Camera Display mode 4 Verify that the following criteria are met 3 5 3 INSTALLATION PROCEDURES TESTING ASSEMBLIES Stream is centered in the illuminated strobe area e Strobe area is centered on the screen Steam and strobe illumination remain centered at all zoom settings Stream remains in focus at all zoom settings Flow cell alignment image is centered on the screen at all zoom settings when camera is in full up position Zoom setting is almost at the mechanical minimum setting 1 0 of the zoom lens when the zoom is in the full out position Zoom or position mechanism does not bind Note If any of these criteria are not met do Heading 4 1 Camera Adjustment Procedure Lasers WARNING Risk of personal injury Laser manufacturers of certain a
440. ou review understand and adhere to the safety precautions for lasers under Heading 1 2 Standard Configuration Procedure Tools Supplies Needed None Procedure l Install the two short targets on the optical plate and verify alignment of the lasers Do not align the lasers unless needed See Standard Laser Alignment Verify back reflection Adjust flow cell as necessary using Flow Cell Alignment Initial Refer to the Headings 3 14 and 3 15 in the Elite ESP Special Procedures and Troubleshooting manual to a Remove the targets reinstall the scatter sensor and use the camera to align the flow cell b Use fluorospheres to align the beam shaping lens focus the laser and the fluorescence detection lens c Optimize adjustments for best CVs and signal amplitudes 4237003 4 1 1 SERVICE AND REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE Standard Laser Alignment Purpose This procedure describes alignment of the standard air cooled Argon and HeNe lasers on the optical table for co linear beams with no other lasers present Refer to the correct laser alignment procedure in this section for all other laser configurations or gated amp alignment Note As a general alignment principle adjust the front laser mount to align to the target closest to the laser Adjust the rear laser mount to align to the target furthest from the laser A series of small iterative adjustments works best Tools Supplies Needed N
441. ound disk type 0 3 located on top of valve bracket assembly 4237003 5 2 1 5 MAINTENANCE PROCEDURES PREVENTIVE MAINTENANCE PMI PROCEDURE 8 Replace the halogen lamp Note The used lamp should be marked as such and left with the operator as an emergency spare 9 Replace the metal holder clamps for both the sheath and waste filters 10 Replace the sheath filter interconnections 11 Replace the sheath filter 12 Replace the waste filter 13 Replace the vial cap 14 Replace all check valves in the waste lines 15 Replace the male and female waste tank connectors 16 Replace the sheath tank cap O ring 17 Inspect and clean a Beam shaping optics b Filters c Mirrors 18 Replace all optical filters used in the filter housing every 5 years 19 Inspect and clean flow cell and fluorescent pickup lens assembly 20 Power up the unit and do the optical alignment procedure 21 At the DOS prompt type DEFRAG and press Note Units with older MS DOS may not have the programs referred to in steps 21 through 25 If not use the Troubleshooter Software 22 there are any problems follow the DEFRAG instructions 23 Reboot the Elite 24 Do Heading 5 1 VERIFICATION INSPECTION PROCEDURE 5 2 2 4237003C 6 SCHEMATICS 6 1 1 6 1 SCHEMATICS REQUIRED 6 1 1 PN 4237003C 6 i 6 ii PN 4237003C 6 1 PN 42370036 SCHEMATICS ER SCHEMATICS REQUIRED This chapter contains a list of
442. output and clamp the line between the regulator and your gauge Watch for the pressure to drop steady drop indicates a leak b Ifyou still cannot find the leak use two clamps one between the regulator and gauge and the other to clamp each leg in turn away from the gauge Activate the Sheath and listen to the sheath regulator Note It should cycle every 10 to 60 seconds to compensate for liquid flow to the flow cell If the regulator does not cycle there may be a leak or internal problem which requires the regulator to be replaced Liquid Leak Testing Sheath Liquid Leak Testing 1 Look for salt residue or rust around fittings and connections at the Sheath Tank Note Salt residue and rust indicate present or past leakage Repair the leak and clean away all deposits Inspect feed thru fittings for small leaks Replace the fittings if you detect a leak PN 4237003CB Figure 7 3 1 Fluidics Troubleshooting Information Fluidics logically troub No Sheath Flow Is Sheath Pressure Present as indicated on Cytometer screen No No Goto No Sheath Pressure Tree Yes TROUBLESHOOTING TROUBLESHOOTING FLUIDIC PROBLEMS eshooting a typical problem No sheath flow MER Is the Sheath pinch valve VL 9 opening Yes 7
443. ow nylon white 0 093 i d to 10 32 flow cell stage 8 2 31 8 2 32 6232215 Fitting brass T 0 250 in o d 6232304 Fitting quick connect panel mount orange 6232305 Fitting quick connect insert orange 6232306 Fitting quick connect insert blue 6232309 Fitting hose quick connect panel mount blue 8 2 3 17 8 2 4 7 6232417 Fitting hose barb 0 062 in i d PVC 8 2 6 6 6232418 Fitting hose barb 0 115 0 180 in i d PVC 8 2 6 4 6232449 Fitting panel mount bottle orange male 6232450 Fitting panel mount bottle blue male 8 2 3 15 8 2 4 13 6232451 Fitting panel mount bottle white male 8 2 3 3 8 2 4 12 6232452 Fitting panel mount bottle yellow male 8 2 3 9 6232472 Fitting quick connect 0 025 od single conn white 6232475 Fitting quick connect 0 125 flow single conn white 6232522 Fitting quick connect 0 375 o d single conn white 6803642 Valve pinch double acting Analyzer 8 2 18 1 6855763 Valve pull apart standard pinch analyzer 8 2 18 12 6856321 Bracket wired sort 6857112 Feet mounting HeNe 8 2 35 5 6857133 Shutter single acting Elite Analyzer 8 2 19 4 8 2 20 2 6858368 Bracket flow cell bimorph sub assembly 8 2 31 6 8 2 32 6 8 1 14 PN 4237003C MASTER PARTS LIST IN NUMERICAL ORDER CATEGORY Table 8 1 10 Hardware Continued PARTS LISTS Part Number Description Figure Item 6858369 Bracket mount Sensor card analyzer 8 2 18 5 6858370 Bra
444. pe C and press Enter Is Windows installed Type NO and press Enter PrintQ Loaded each Boot Type YES and press Enter Press Enter Type the PrintQ serial number in the highlighted area You can find the serial number on the rear cover of PrintQ Users Guide amp Reference Manual Press Enter Type the requested information when prompted for NAME press Enter TITLE and press Enter PHONE and press Enter COMPANY and press Ente ADDRESS and press Enter CITY and press Enter 4 9 1 4 SERVICE AND REPAIR PROCEDURES SOFTWARE INSTALLATION 4 9 2 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 4 41 42 43 44 45 46 47 48 49 Press Enter When PrintQ programs have been successfully installed press Ente Select N for No Auto Prompting Select 1 for number of printers connected to computer and press Enter Select Printer 1 Printer Port LPT1 and press Enter Use to select the customer s Printer type then press Enter To accept Printer setup press Ente Type N for Do Not Run Test Type N for Do Not Print Registration Form Remove all diskettes from drives Press Ctrl Alt Delete to boot up the Elite System Software Press Enter Press F2 Enter to exit software and return to DOS Press cu t amp P to enter the PrintQ Status Screen Press At 2
445. personal injury l Ensure that the Cytometer is removed from the delivery truck with a fork lift or pallet carrier Inspect all boxes for damage Notify shipping of any external damage Remove the banding and cardboard cover from the Cytometer Unbolt the wooden bracket that secures the Cytometer to the pallet as shown in Figure 3 2 1 CAUTION Risk of instrument damage Be sure to properly secure the special ramps with the clips provided Do not use the U shaped clips Use the flat clips 5 Place the ramps in front of the Cytometers pallet See Figure 3 2 1 Figure 3 2 1 Unloading the Cytometer PALETTE 7003085A PN 4237003 3 2 1 INSTALLATION PROCEDURES INITIAL SETUP 3 2 2 Secure the ramps to the pallet with the flat drop in clips as shown in Figure 3 2 1 Verify that the ramps are properly aligned with the pallet Roll the Cytometer on its built in casters down the ramps and to the desired installation location Note The Cytometer requires a moving clearance of 143 cm 56 in by 77 cm 30 in by 130 cm 51 in If the Elite configuration includes an optional water cooled laser Innova 300 instruct the customer to store the crates in case the laser has to be sent back to the factory for service Note The specifications of the packed laser are the following Power Supply crated gt Length of 91 cm 36in Width of 74 cm 29 in Height of 48 cm 19 in gt Weight of 28 kg 62 Ib
446. plane Each circuit card recognizes one or more addresses and responds by reading the associated data word The data word controls a particular function on the card with each circuit card representing one or more ports to the CPU The implementation of the functions is more complex however Although the cards on the multibus decode the complete address cards in the upper and lower card cages are addressed less directly The Data Taker Interface card recognizes the range of addresses corresponding to the upper and lower card racks The system then routes the partially decoded addresses to the Gated Amp Control card which recognizes the addresses unique to the upper backplane Interface and Scaler card which recognizes the addresses unique to the lower backplane Data Taker Interface Card The Data Taker Interface card e Connects the Multibus subsystem to the three card racks in the right pedestal The upper card rack is the Gated Amp card cage The lower card rack is the Acquisition card cage The racks in back of the unit are the PMT power supply card cages e Provides the CPU with the connection to the Gated Amp Control card to control all cards in the Gated Amp card cage e Provides the CPU with the connection to the Sensor Interface card to send commands to the Pneumatic Interface card and the Motor Control card e Provides the CPU with the connection to pass commands through the Interface and Scaler card to control all cards
447. ple line o o Yes Sample line Sample Insertion rod r Sample pickup tube is clogged ean or replace Mix Sample liquid vigorously to No form bubbles Do the bubbles compress when you press Run Does i the sample tube level go down when in Run Sample Pressure is not present or incorrect Go to Fluidics troubleshooting Go to No Fluidics Troubleshooting Fluidics OK v Possible problem with YES Do histograms acquire on the Lister Out Card Cytometer screen Lister AB Card Parallel Cables between lister cards NO NO Do pulses appear on the isi i __ Problem is in analog signal Cytometer Scope screen path Use your oscilloscope to trace signal paths to YES locate problem Do dots appear on the Cytometer NO Possible problem with Scattergram screen Mux amp Scope Card or PSH Card Use your scope to test YES Is there a non zero data rate Possible problem on the Cytometer Main screen with ADC Card Possible problem with Lister Out Card 7003079A 7 2 2 4237003C TROUBLESHOOTING TROUBLESHOOTING THE ACQUISITION SUBSYSTEM 7 Acquisition Troubleshooting Methods Subtraction Color Compensation Rule out electronic problems by testing the electronic subtraction circuits for proper operation If no electronic problem is detecte
448. ply Charges the Deflection plates Only active when turned on by Sort Output card 36 V Power Supply Provides power to the Sort Oscillator card for bimorph oscillation and stream deflection 90 V Power Supplies 2 Two identical 90 V power supplies are used to power the Sort Output card One supply provides positive output and the other provides x90 V grounded negative output to the card The supplies also provide power to the 36 V Regulator card which provides 36 V to the Sort Output card 15 V 5 A Power Supply Provides power to the card cage and to the motor controller for the camera 15 V 3 4 A Power Supply Provides power to the DACs for the PMTs 15 V 0 25 A Power Supply Provides power to the PMTs and the FS Detector 5 V Power Supply Provides power to the KIR3 card to make selections on the Control Screen and on the CPU 12 V Power Supply Provides power to the dual CRTs and to the CPU 24 V Power Supply Provides power to the solenoids PN 4237003C 2 8 19 2 INSTRUMENT DESCRIPTION SORT SUBSYSTEM Figure 2 8 12 Power Supplies Location FIBER OPTICS SUPPLY PMT HV SUPPLY 36 V SUPPLY circuit card SWITCHER SUPPLY FAN 107 CFM 115 VAC DIE Wat Did DO 3000 DEFLECTION SUPPLY 15 V 0 25 SUPPLY 15 V 5 0 A SUPPLY 90 V SU
449. positions as shown in Figure 4 12 12 Figure 4 12 12 Laser Beam Centered Across Forward Scatter Detector 7003199A 10 Verify that pulses appear when you scan between the horizontal blue line positions 11 If no pulses appear or if they do not appear to be normal a Check for a fluidics problem such as bubbles tube not sealed and so forth b Verify that the flow cell tip is clean Clean if needed c Verify that the forward scatter mask is all the way in position PN 4237003 4 12 7 SERVICE AND REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE 12 If pulses still do not appear or if there is a lot of background noise scattered light from the beam may be entering the sensor a Adjust the laser beam horizontal knob so the beam is in the center of the mask and all noise disappears from the scope b Adjust the flow cell Z axis knob until good forward scatter pulses appear See Figure 4 12 13 for knob location 13 Maximize the forward scatter pulse by adjusting the flow cell Z axis knob 14 Fine tune for maximum forward scatter pulse height by adjusting the horizontal laser beam knob 15 Do Optical Fine Tuning Figure 4 12 13 Flow Cell Z Axis Knob Location FLOW CELL LASER BEAM INTERLOCK VERTICAL VERTICAL DEFEAT BAR ADJUSTMENT ADJUSTMENT FLOW CELL ME Z AXIS gt ADJUSTMENT FLOW CELL HORIZONTAL ADJUSTMENT 5 LAGER N _ S HORIZONTAL um 9
450. programmable single cell deposition system that e Sorts a user defined number of cells per well into multi well microculture plate e Automates hybridoma screening for research and development of monoclonal antibodies sterile sorting of bacterial cultures and screening for high density monoclonal markers e Accepts standard 6 12 24 48 60 or 96 well microculture plates for automated cell deposition e Automates the process of determining the optimum Drops Delay setting through its Sort Matrix function Figure 2 2 1 Autoclone Sorting Option Mechanism lt ol gt E i gt Ld _ 227 7717 esch 7003149 Sorting Tray Wells The Autoclone Sorting Option steps through the wells of the tray in a serpentine path skipping unprogrammed wells For example in a 24 well tray the first well will be A1 followed by A2 A4 A5 B6 B5 B4 B3 B2 B1 C1 C2 C3 C4 C5 C6 At this point the plate is as far to the left as possible therefore the Autoclone Sorting Option rotates the tray 180 degrees and then continues to sort until the tray is finished Preventing Electrostatic Build Up Electrostatic build up in the sort wells is prevented because the Autoclone Sorting Option does not charge droplets containing target cells The waste droplets are charged and deflected into the waste catcher and only uncharged droplets containing the desired cells fall into the sort wells PN 423
451. properly aligned Do alignment procedure FS pulse shifted mean high CVs fluorescence okay Dirty flow cell Clean flow cell with lens cleaning paper cotton swab and optical grade methanol FS detector mask not positioned properly Reposition mask Drain s overflow Waste tank not sealed Reseal tank Quick disconnects not connected Reconnect quick disconnects Pinched waste line Follow drain line and remove pinch Weak vacuum pump Acquisition appears on the cytometer screen but not on the workstation monitor Problem with Lister card or with parallel cable Table 7 1 3 General Troubleshooting Information Problem Probable Cause Corrective Action Autoclone Sorting Option Noisy movement Incorrect motor speed Adjust motor speed Interference or friction Locate and correct Unit hits limit switch during calibration Optical sensor 1 Check optical sensor and encoding strip for dust or contamination Wipe clean as needed 2 Verify operation of sensor by manually moving the arm assembly while reading the marker box on the Autoclone Sorting Option Control screen PN 4237003C 7 TROUBLESHOOTING ISOLATING THE PROBLEM AREA Table 7 1 3 General Troubleshooting Information Continued Problem Probable Cause Corrective Action Offsets too much Realign the mechanism to center the uncharge
452. pulses do not appear normal Optics are not aligned Do Heading 4 1 OPTICAL ALIGNMENT PROCEDURE Flow cell tip is dirty Clean the flow cell tip Forward scatter mask is not properly in place Verify that the forward scatter mask is properly seated Scattered light from the beam may be entering the sensor Adjust the beam horizontal to the center of the mask until all noise if any disappears from the scope 2 Adjust the flow cell Z axis until good forward scatter pulses appear 3 Roughly maximize the forward scatter pulse with the flow cell Z axis 2 signals low and require increased HV to be seen properly Flow cell is not aligned to the center of the fluorescence pickup lens Adjust the flow cell vertical and or horizontal until the pulse increases PN 4237003C 7 TROUBLESHOOTING ISOLATING THE PROBLEM AREA Table 7 1 5 Optics General Troubleshooting Information Continued Problem Probable Corrective Action Traces were lost while setting up the scope Pulse selection for Trace 1 is too low Select FS for Trace 1 initially to establish pulses Check desired Trace 1 pulse on Trace 2 and adjust to a reasonable pulse height before displaying on Trace 1 3 Select traces in order With a 4PMT system select Trace 1 then PMT2 Peak and Trace 2 and PMT4 Peak With a 5PMT system select Trace 1 then PMT2 Peak and Trace 2
453. quency gives the highest breakoff but when the frequency is adjusted 0 196 up or down the last attached drop moves downstream and becomes very unfocused PROCEDURE The bimorph has not sufficiently warmed up Allow the bimorph to warm up at least 30 minutes before determining the correct crystal frequency There is a clog or bubble in the flow cell tip and or body Press CLEAR three times then press DEBUBBLE Clean the flow cell Insufficient droplet breakoff with 90 crystal drive When crystal drive is 90 or greater the droplet breakoff is still too far downstream which means it is still below the ground plate when all the way down Hydraulic or pneumatic leak or problem 1 Check the sheath tank cap sheath filter sheath tubing through the system Use soapy water to find the leaks when bubbles occur 2 Check that both the sheath and sample regulators are working properly Rapid occurrence of the regulator cycling lt 5 to 10 seconds between repetitive cycling indicates a problem Flow cell or body is not tightened properly Verify that the flow cell and body are properly tightened Flow cell is clogged or cracked Remove and clean flow cell Inspect before replacing If cracked replace There is a bubble in the flow cell Press CLEAR three times then press DEBUBBLE Double side streams There is a second side stream between the correct stream
454. quency to 32 kHz Connect the oscilloscope to the alligator clip that attaches to the flow cell s sample insertion rod Note You can leave the clip connected to the flow cell Turn on a Sort Test b Sort Left Enable c Sort Right Enable Set Sort Right and Left Stop counters to OFF Press START and verify the sort counters are counting the Sort Test pulses Set Deflection to 100 42370036 10 11 12 13 SERVICE AND REPAIR PROCEDURES SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE Set Drops Sorted to 3 Adjust Stream offset so the signal baseline is 0 Vdc Measure amplitude of the center level from the baseline and verify the value is 80 V 22 5 V If the value is incorrect do Sort Pulse Amplitude Adjustment If the value is correct go to step 10 Verify that the sort pulse appears as shown in Figure 4 2 1 for 1 2 and 3 drop sorts Verify that the negative going pulse is symmetrical with positive going pulse Adjust front and back porches to ensure they can be varied from zero to 10096 Return front and back porches to reference values e Front porch at 80 e Back porch at 20 Sort Pulse Amplitude Adjustment D ON DM kW Hr pg 13 Set Drive Frequency to 32 kHz Turn on Sort Test Turn on Sort Left Enable Turn on Sort Right Enable Set Sort Right and Left Stop counters to OFF Press START and verify the sort counters are counting the Sort Test pulses Set Deflection to
455. r prism 8 2 19 10 6857598 Ring jet in air 8 2 16 2 6857636 Adapter interlock bypass 8 2 16 5 6857637 Bracket interlock relay subassembly 8 2 16 6 PN 4237003C 8 PARTS LISTS MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY Table 8 1 6 Cytometer Continued Part Number Description Figure Item 6858907 Door sample drawer Analyzer 8 2 17 13 6858908 Drain sample sample drawer 8 2 17 10 6858923 Cap sample tube 8 2 17 2 Table 8 1 7 Filters Part Number Description Figure Item 1015831 Biohazard filter container 3802050 515 nm LP laser blocker filter 3802073 530 SP filter 3814044 2 0 ND filter 3814069 635 nm BP 3814222 beam splitter 633 nm 3814227 395 nm BP filter air cooled laser 6232143 Hydrophobic round disk 0 3 micron filter 8 2 5 10 6232473 Sheath 0 2 micron 8 2 3 18 6232489 Waste 0 2 micron 8 2 3 14 6857204 Holder filter 6857205 675 nm BP filter 3814139 6857206 575 nm BP filter with holder Elite ESP 3814135 6857207 600 nm DL filter Elite ESP 3814138 6857208 550 nm DL filter Elite ESP 3814067 6857209 525 nm BP filter Elite ESP 3814134 6857210 488 nm BK filter Elite ESP 3802072 6857211 488 nm BP filter Elite ESP 3814137 6857212 488 nm DL filter Elite ESP 3814136 6857370 Mirror filter 3805018 6857500 1 0 ND filter and 530 SP assembly 3802048 3802073 6858219 381 nm BP filter HeCd laser 3814264 6858220 440 nm DL 3814199 6858249 633 n
456. r Mux slot 13 J14 PN 42370036 PN 42370036 ELECTRONIC ADJUSTMENTS CABLE AND JUMPER CONFIGURATIONS Table 4 10 6 Cable Connections Continued SERVICE AND REPAIR PROCEDURES From To Mux and Scope 27 in cable J10 Scat CV slot 14 J4 Mux and Scope 27 in cable J11 Scat CV slot 14 J3 Mux and Scope 27 in cable J12 Scat CV slot 14 J2 Mux and Scope 27 in cable J13 3 PMT Sub SW R1 slot 5 J8 Mux and Scope 27 in cable J14 3 PMT Sub SW R1 slot 5 J7 Mux and Scope 27 in cable J15 Peak Scatter Mux slot 13 J4 Mux and Scope 27 in cable J16 Dual FL slot 3 Mux and Scope 27 in cable J17 Dual FL slot 3 Mux and Scope 27 in cable J19 J3 Dual FL slot 3 J4 J2 J7 Dual FL slot 3 Mux and Scope 27 in cable J20 Dual FL slot 3 J8 Mux and Scope 27 in cable J21 Scat CV slot 14 J9 Mux and Scope 27 in cable J22 3 PMT Sub SW R2 slot 4 J8 Mux and Scope 27 in cable J23 3 PMT Sub SW R2 slot 4 J7 Mux and Scope 27 in cable J24 Peak Scatter Mux slot 13 J10 Mux and Scope 27 in cable J25 Gated Amp Control R3 slot 1 J2 Mux and Scope 27 in cable J18 Mux and Scope 27 in cable J31 Gated Amp Control R3 slot 1 J6 Gated Amp Units Interconnections from Sensor Area FALS WM400 S
457. r and can be read when the holder is installed in the filter block Long Pass LP Filters LP filters absorb short wavelength light and pass long wavelength light They are identified by their 5096 transmittance wavelengths Do not use these filters as the only means to block light Short Pass SP Filters SP filters pass short wavelength light and reflect long wavelength light They are identified by their 5096 transmittance wavelength Blocking BK Filters BK filters pass light at all wavelengths except for a narrow band of blocked wavelengths The filters are identified by the laser line they most effectively block they typically pass about 0 01 to 1 096 of those wavelengths Use BK filters in the detector compartment to block intense laser light scatter Band Pass BP Filters BP filters pass a narrow band of wavelengths and block all others Use these filters to pass fluorescence light from a single dye while blocking light from other dyes Dichroic Long Pass DL Filters DI filters pass longer wavelengths and reflect shorter ones These filters are identified by their 5096 transmittance wavelength Use these filters in diagonal positions PN 4237003C PN 42370036 INSTRUMENT DESCRIPTION OPTICAL COMPONENTS Dichroic Short Pass DS Filters DS filters pass shorter wavelengths and reflect longer ones DS filters are not available through Beckman Coulter Neutral Density ND Filters ND filters do not discriminate l
458. r base is not in place or sensor is defective Autoclone Sorting Option Card The Autoclone Sorting Option card interfaces between the Cytometer multibus CPU and the Autoclone Sorting Option mechanism Through this card the CPU operates the mechanism motors to move the tray and read the optical position sensors and limit switches This is not an intelligent microprocessor equipped card The system CPU must issue an command for each step of the stepper motors The card converts a command from the CPU into the appropriate four phase current to move one of the stepper motors The status of the optosensors and the limit switches are encoded and placed on the databus for the CPU to read The status of the optosensors and limit switches are also displayed by LEDs on the card Note that the limit switches are latched on the card Once a limit switch has been activated the error condition remains until the card is reset This is done by recalibrating the Autoclone Sorting Option Autoclone Sorting Option Interconnect Card PN 4237003C The Autoclone Sorting Option Interconnect card is attached to the rear of the mechanical assembly and serves as the connection point for the cable to the Cytometer The card contains no active components but provides connectors for the two flex cables and a connection to the X motor front back Position Detection Cards Two Position Detection cards are used in the mechanism one is mounted to the X moveme
459. r before proceeding with the sort or the purity and recovery may be affected 6 Using the settings obtained above to sort a mixture of different brightness beads right and left 50 000 each direction non ESP systems sort at 1 500 events sec data rate with Abort ON For ESP systems sort at 5 000 events sec with Abort ON Complete Abort ON 7 Reanalyze the sorted beads to determine purity e For non ESP purity is gt 95 e For ESP purity is gt 99 Sort Coincidence Verification This procedure verifies operation of the sort coincidence circuitry Perform this procedure as follows e After you replace a Sort Oscillator Pulse Pileup Det TOE or Sort Delay card When operation of the coincidence circuits are in question Verification and test of the coincidence circuitry is done by simulating the four possible coincidence events that can occur Good followed by good Good followed by bad Bad followed by good e Doublets where the cells are too close together to be resolved as separate cells Note Doublets are a function of the PPU Note The PPU parameter Par 1 also generates a POSITIVE PPU DETECT signal if that parameter is above 9 8 V or greater than channel 1 000 These simulations verify the operation of the four coincidence functions Sort pulse extension Sort pulse truncation e Abort of two events in same droplet Extended abort To verify the operation of the four coincidence
460. r from the Cytometer Control Valves screen e With Autoclone Sorting Option waste OFF the waste catcher is retracted e With Autoclone Sorting Option waste ON the waste catcher is extended With the waste catcher retracted adjust the Deflection Gain and move the waste catcher from left to right until the left side stream the one flowing straight down does not hit the waste catcher and the waste stream enters the waste catcher cleanly Verify that the waste catcher is correctly positioned so the extended and retracted positions function correctly Note Loosen the clamp holding the air cylinder which moves the waste catcher and adjust as needed Operational Test Many factors affect the outcome of this test the presence of liquid at all well locations and particles in some wells indicates Autoclone Sorting Option function Actual results depend on the condition and adjustment of the host instrument ge Prepare a sample of fluorospheres and place a sort region around the fluorospheres Configure the system to sort 1 particle into each well of a 96 well plate Place the cover of the well plate into the Autoclone Sorting Option and sort Check the plate cover with a microscope to confirm sorting 3 10 9 INSTALLATION PROCEDURES AUTOCLONE SORTING OPTION INSTALLATION 3 10 10 PN 4237003 INSTALLATION PROCEDURES 5TH PMT KIT WITH SWITCHABLE AMPS INSTALLATION PROCEDURE 3 11 5TH PMT KIT WITH SWITCHABLE AMPS INSTALLATION
461. r the cards in the Gated Amp card cage as shown in Table 4 10 4 4 10 2 PN 4237003C ELECTRONIC ADJUSTMENTS CABLE AND JUMPER CONFIGURATIONS Table 4 10 4 Gated Amp Card Cage Jumper Settings SERVICE AND REPAIR PROCEDURES Slot Card Jumpers 1 Gated Amp Control R3 None 3 Dual FL SW R E8 E9 E11 E12 4 3 PMT Sub SW R2 4 9 11 5 Sub SW R1 9 11 6 Gated Amp 7 20 40 60 Microsecond Delay 2 E1 E2 4 E10 E11 In 8 7 Microsecond Delay 2 E10 E15 In 9 7 Microsecond Delay 1 E10 E11 In 10 20 40 60 Microsecond Delay 1 E1 E2 E3 E4 E9 F10 In 11 Scat Aux Gated Amp 12 3 PMT Sub SW R E1 E2 E5 E6 E9 E11 13 Peak Scatter Mux SW R E1 E2 E3 E4 5 7 14 Scat CV SW R1 E9 E11 E5 E7 16 Sensor Interface E2 E3 E5 E6 E8 E9 E11 E12 E13 E14 E16 E17 E19 E20 and E22 E23 Data Acquisition Card Cage Configuration Jumper the data acquisition card cage cards as shown in Table 4 10 5 Table 4 10 5 Data Acquisition Card Cage Cards Jumper Settings Slot Card Jumpers 1 left Pulse Generator and Clock R X1 through X4 5 Pulse Pileup Det TOF E1 E2 E20 E21 6 Mux and Scope Interface E1 E2 E3 E4 7 Quad PSH 1 E2 E4 8 Quad PSH 2 E2 E4 9 Peak ADC PSH Control E1 E2 E6 E7 10 Lister Out E5 E6 11 Prism and Sort Window Test None 12 Bitmap and Sort Decision None 13 Interface and Scaler R E1 E5 E2 E6 E3 E7 14 Sort Delay R3 None PN 4
462. r to Dual CRT pivot mount 4 10 5 electronic 3 4 1 electronic illustration of 3 4 1 for 5 PMT installation 3 11 1 3 11 2 for all instrument configurations 4 10 5 for Gated Amp instruments 4 10 6 for non Gated Amp instruments 4 10 6 for pulse generator setup 4 2 10 for Sort ESP systems 3 13 4 HeCd laser power supply illustration of 3 7 7 interconnections from Sensor Area for Gated Amp instruments 4 10 9 interlock cable 3 7 4 Mux and Scope card 4 10 5 Quad PSH 1 card 4 10 5 to Workstation CPU 3 4 2 to Workstation CPU illustration of 3 4 2 cables back panel wiring configuration 4 10 1 blue ribbon connections of 4 10 4 coax Mux and Scope card function of 2 13 2 DIN Camera Interface card 2 5 5 various configurations of 4 10 1 calibration of Autoclone Sorting Option 2 2 4 camera adjustment procedure 4 1 9 how to aim 4 1 9 how to focus 4 1 10 how to optimize the image 4 1 11 how to position movement 4 1 11 how to zoom 4 1 10 initial setup procedure 4 2 7 pan feature function of 2 5 5 video output 2 5 5 camera image optimization See camera INDEX 3 INDEX Camera Interface card cable DIN function of 2 5 5 camera image R99 adjustments for best 4 1 11 function 2 5 5 jumpers 4 10 2 A 2 2 camera movement See camera capacitors 22 uf replacing R77 with 3 17 1 diode charged through 2 13 4 function of 2 6 5 integration function of 2 6 5 card cages Acq
463. r to Solenoid Valves The system vents sample sheath and rinse tank pressures into the drip chamber The drip chamber is vented to the atmosphere through a biofilter If this filter clogs up then pressure builds up in the drip chamber and the drip chamber switch detects the buildup On the screen this appears as Drip Chamber Error The system draws air from the waste bottle through the waste filter The water filter is a hydrophobic filter that clogs if the waste bottle overflows This condition results in an increase in vacuum detected by the waste filter switch On the screen this appears as Waste Filter Error Solenoid Valves Each solenoid valve performs a specific function See Table 2 9 1 for a description of manifold valves 1 15 See Figure 2 9 1 for an illustration of valve operations PN 42370036 2 9 1 2 INSTRUMENT DESCRIPTION PNEUMATICS SUBSYSTEM Table 2 9 1 Solenoid Valves Manifold Number Name Type When Activated VL1 System Pressure Solenoid Allows flow of air to the sheath and sample pressure regulators when energized VL2 Laser Shutter Solenoid Allows flow of air to laser shutter cylinder Allows laser to pass when energized VL3 Sample Pinch Solenoid Provides air to the sample pressure pinch valve actuator which activates the pinch valve to close the sample tubing VLA Sample Pressure Solenoid operated Passes regulated sample pressure to the sample tube
464. rary version 4 02 OR Elite Pentium PC Upgrade Kit 6915006 which includes gt Elite Intel Pentium computer with SVGA video board and 16 MB RAM gt Serial mouse gt Elite software library version 4 02 Procedure ATTENTION Software version 4 02 is for use with Pentium computers only Do not install version 4 02 on anything other than a Pentium computer 1 Verify that the ac power setting on the rear of the computer matches local ac power on site Install software version 4 02 according to the README DOC file supplied with software Remove the Lister A B card from the old computer and install it in slot 1 of the new computer as viewed from the front 3 21 2 4237003C PN 42370036 Verify the following cable connections COM1 COM2 e Mouse Opin serial cable Lister cable e Keyboard cable e Monitor keyboard port Reboot the computer by pressing Alt Delete Note Do not change the BIOS settings unless absolutely necessary To change them press during the boot up process to access the BIOS Setup Menu The system BIOS is configured as follows Standard Setup FLOPPY A FLOPPY B MASTER DISK SLAVE DISK Advanced Setup TYPEMATIC RATE PROGRAMMING SYSTEM KEYBOARD PRIMARY DISPLAY MOUSE SUPPORT ABOVE 1 MB MEMORY TEST MEMORY TEST TICK SOUND PARITY ERROR CHECK HIT DEL MESSAGE DISPLAY EXTENDED BIOS RAM AREA WAIT FOR F1 IF ANY AREA SYSTEM BOOT UP NUM LOCK FLO
465. rator location with Enterprise laser illustration of 3 14 3 beams alignment of 4 12 1 4 12 5 calibrating for Gated Amp 3 9 24 collinear alignment procedure 3 12 4 collinear laser alignment for three lasers 4 1 6 configurations possible 3 7 1 Enterprise laser installation 3 14 1 Enterprise laser mounting illustration of 3 14 3 hazards of laser beams 1 2 2 HeCd optical arrangement illustration of 3 7 8 installing HeCd covers 3 7 12 installing laser covers water cooled 3 7 13 installing optional third laser 3 7 1 INDEX 9 INDEX mounting procedure 4 1 4 mounting procedure during beam translator upgrade 3 12 1 radiation hazards 1 2 2 specifications summary of 2 3 2 standard alignment procedure 3 12 1 technical specifications 2 3 3 test procedures 3 5 4 time space separated laser 4 1 7 time space separated laser alignment procedure 3 12 5 warning labels in sensing area 3 7 14 warning labels on rear of instrument 3 7 15 water cooled laser illustration of 3 7 10 water cooled optical arrangement illustration of 3 7 9 leaks air testing for 7 3 2 sheath testing for 7 3 2 sort current leakage test 7 5 1 7 6 1 LEDs KIR3 card function of 2 5 5 parabolic mirror image in 4 12 4 lens block location 4 12 2 lenses See optical elements Lensmaker s equation 2 7 4 light defined for optical elements 2 7 1 infrared IR defined 2 7 1 refraction defined 2 7 1 travel media used 2 7 1 wavelengths
466. re 3 9 22 laser configuration 3 9 22 PMT 2 adjusting 3 9 17 PMT 3 adjusting 3 9 17 PMT 4 adjusting 3 9 18 PMT 5 adjusting 3 9 18 potentiometer adjusting 3 9 10 Gated Amp card function of 2 12 2 Gated Amp Control card block diagram 2 12 1 function 2 5 6 2 12 1 Gated Amp Control R2 card jumpers 4 10 2 A 2 2 Gated Amp Control R3 card function 2 12 2 installation 3 9 3 jumpers 4 10 3 A 2 3 gauge pressure See pneumatics generator gate window activation of 2 12 2 H house air installation 3 8 1 8 INDEX HP 1200C color printer replacement procedure 3 20 1 HP 1600C color printer cartridges part numbers of 3 20 1 editing CGI CFG file for DOS 6 0 and higher 3 20 2 editing CGI CFG file for lower than DOS 6 0 3 20 4 HP 1200 C replacement of installation procedure 3 20 1 model numbers affected instrument 3 20 1 testing after installation 3 20 6 HV DAC 2 card jumpers A 2 1 HV DAC card jumpers 3 11 1 A 2 1 HV DAC Control card jumpers 4 10 4 A 2 1 Hx marker See Autoclone Sorting Option 2 2 3 Hy marker See Autoclone Sorting Option 2 2 3 hydraulics See pneumatics image formation description 2 7 2 infrared IR defined 2 7 1 inputs Vac laser auto transformer 100 Vac to 4 3 1 laser auto 115 Vac to 4 3 1 laser auto 230 Vac to 4 3 2 laser auto 240 Vac to 4 3 2 main system transformer 100 Vac to 4 3 1 main system transformer 115 Vac to 4 3 1 main system transformer 230 Vac
467. re 7 1 1 General Troubleshooting 7 1 2 7 2 TROUBLESHOOTING THE ACQUISITION SUBSYSTEM 7 2 1 Purpose 7 2 1 Tools Supplies Needed 7 2 1 Procedure 7 2 1 Acquisition Troubleshooting Methods 7 2 3 Subtraction Color Compensation 7 2 3 PMT 3 PMT 2 Compensation 7 2 3 Log Amplifiers 7 2 4 7 5 TROUBLESHOOTING FLUIDIC PROBLEMS 7 3 1 Purpose 7 3 1 Tools Supplies Needed 7 3 2 Air Leaks 7 3 2 Sheath Air Leak Testing 7 3 2 Liquid Leak Testing 7 3 2 Sheath Liquid Leak Testing 7 3 2 7 4 TROUBLESHOOTING PRESSURE CONTROL PROBLEMS 7 4 1 Purpose 7 4 1 Tools Supplies Needed 7 4 1 Procedure 7 4 1 Sheath Pressure Problems 7 4 1 Sample Pressure Problems 7 4 3 7 5 TROUBLESHOOTING THE SORT SUBSYSTEM 7 5 1 Purpose 7 5 1 Tools Supplies Needed 7 5 1 Sort Current Leakage Test 7 5 1 7 6 RESETTING CYTOMETER SOFTWARE LOCKUPS 7 6 1 Purpose 7 6 1 Tools Supplies Needed 7 6 1 Reset the Cytometer CPU 7 6 1 Clear the CPU s Memory on the Cytometer 7 6 1 ILLUSTRATIONS 7 2 1 Acquisition Troubleshooting Flowchart 7 2 2 7 2 2 Filter Configuration 7 2 3 7 3 1 Fluidics Troubleshooting Information 7 3 3 7 4 1 Troubleshooting Fluidics for No Sheath Flow 7 4 1 7 5 1 Troubleshooting Sort Problems 7 5 1 7 5 2 Satellite Droplets 7 5 8 4237003 7 1 7 TABLES 7 1 1 7 1 2 7 1 3 7 1 4 7 1 5 7 1 6 7 1 7 7 1 8 7 3 1 7 5 1 Isolating Problem Areas 7 1 1 Autoclone Sorting Option Gener
468. re Item 1016430 Restrictor tubing 2 0 in 1016487 Restrictor tubing 7 0 in 8 2 20 12 3202039 Waste carrying tubing 0 145 i d 3203016 Pinch valve tubing silicone blue stripe 0 079 i d x 0 031 3210004 Sample line tubing 0 010 i d x 0 036 8 2 17 3 8 2 19 11 8 2 20 6 Table 8 1 23 Workstation Part Number Description Figure Item 1018377 Pen graphics pad 1019318 Clip AT 1022173 Mouse pad 2016161 Mouse serial 8 2 2 1 2016279 Cartridge disk removeable Q PAK 2016280 Drive 5 25 in remoeable 2016297 Disk controller Profile XT 2016298 Drive floppy 5 25 in 1 6 MB 8 2 28 5 2016328 Drive floppy 3 5 in 8 2 28 5 2016340 Disk controller Profile AT long board 2016368 Cartridge ribbon Fujitsu printer 2016381 Drive floppy 5 25 in 1 2 MB 2016383 Power supply switching 115 Vac 60 Hz 230 Vac 120 Hz 2016387 Workstation Intel 301Z 2016392 Keyboard enhanced 102 key 2016413 Card video VGA SW 2016416 Drive non removable 85 32 MB 8 2 28 3 2016419 Card Adapter PC to Ethernet 2016424 Card Disk Controller 1003 V 8 2 28 6 2016426 Drive optical 600 MB 115 Vac 2016427 Pack Cartridge optical disk 2016447 Workstation Intel 302 2016457 Card adapter SCSI host 2016458 Drive disk cartridge SCSI 20 MB PN 4237003C 8 1 23 8 PARTS LISTS MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY Table 8 1 23 Workstation Continued Part Number Description Figure Item 20
469. re within specification for the fluorospheres used Verify operation of the oscilloscope Verify operation of the GRAPH display on the Cytometer Operational Test Sorting Ability If repairs were made affecting sorting electronics or mechanical elements do Heading 4 2 SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE Operational Test Every Call 1 2 Have customer run one or more samples to verify operation Ensure that all customer concerns have been addressed TOF Verification Purpose This procedure describes how to verify TOF by creating a simple protocol to measure the FL Test pulse width using the PMT2 Peak signal Procedure 1 At the Workstation a Select Acquisition Parameter b Erase all parameters that may be present Verify that you are in the Create mode Select the following parameters in the Signal column a 2 b PMT2 Peak c Time of Flight d 2 Peak Note The list of Parameters in the right column should read PMT2 PMT2 Peah Time of Flight The Time of Flight box should contain Param PMT2 Peak these items are not correct repeat step 3 PN 4237003C 10 11 12 13 PN 42370036 MAINTENANCE PROCEDURES VERIFICATION INSPECTION PROCEDURE At the Cytosettings screen a Setthe Time of Flight time base to 10 0 microseconds b Setthe PMT2 Peak discriminator to 100 other channels to OFE At the Protocol screen create single parameter histograms
470. rewdriver or use crew retaining Phillips head screwdriver b Bring the screw to the connector inside the Motor Controller card cage and fasten the screw to the threads in the connector mounting block Install the Motor Controller card in the bottom slot If the instrument is not Autoclone Sorting Option Ready from the factory do Upgrading the Instrument to be Autoclone Sorting Option Ready Upgrading the Instrument to be Autoclone Sorting Option Ready Do this procedure if the instrument is not Autoclone Sorting Option ready 1 Install the Autoclone Sorting Option card into the empty slot above the Motor Controller card Open the pneumatic housing door Remove the screw plugs from the 13th and 14th valve positions second and third positions up from the bottom of the pneumatic manifold Disassemble the new solenoid valves noting the position of the internal assemblies Install solenoid valve bottoms at positions 13 and 14 and assemble the solenoid valves Fasten terminals with WM 342 and WM 343 on the adjoining cable assembly to the solenoid valve at position 14 Fasten terminals with WM 340 and WM 341 on adjoining cable assembly to the solenoid valve at position 13 Install a fittings with a seal on the manifold at a right angle to the solenoid valve as position 14 3 10 3 INSTALLATION PROCEDURES AUTOCLONE SORTING OPTION INSTALLATION 3 10 4 10 11 12 13 14 15 16 1T 18 19 20
471. rform the various subtraction combinations required You can verify the electronic operation of the subtraction circuit by providing the same input to the main channel and the channel to subtract The main channel is reduced by the percent of signal selected for subtraction Note The Elite performs the subtraction on peak signals This means that the subtraction percentages directly correlate with peak signal channel numbers but do not correspond for integral or log Log Amplification The Elite uses log amplification to compress very wide distributions of information to fit conveniently on a histogram display Using four decade amplifiers the Elite can accommodate a 10 000 1 range of intensity between the first and last channels of a histogram Figure 2 6 2 shows the response of the log amp to the input voltage 2 6 2 PN 4237003C INSTRUMENT DESCRIPTION 2 ACQUISITION SUBSYSTEM Figure 2 6 2 Log Amp Response 0 8 Voltage 6 Out of Log Amp 4 2 0 0 001 2 3 4 5 6 7 8 9 10 Voltage Into Log Amp 7003033A In Figure 2 6 2 the output is for an input of 1 mV 0 001 V This corresponds to channel 0 on the histogram Any input voltage less than this including 0 V results in a negative output from the log amp Using the following formula the amp output would approach a very large negative number as the voltage approaches 0 The Elite s circuitry includes a cla
472. riminator channel use one of the other two settings Note The minimum acquisition window length is 5 microseconds even if the triggering pulse is shorter than 5 microseconds e 5 microseconds The acquisition window begins at the first discriminator crossing and remains open for 5 microseconds after that time This setting is appropriate for sense in air flow cell high bandwidth operation e 20 microseconds The acquisition window begins at the first discriminator crossing and remains open for 20 microseconds after that time This setting is appropriate for sort sense flow cell operation Figure 2 13 8 shows the timing PN 4237003 213 11 INSTRUMENT DESCRIPTION SIGNAL SELECTION AND DIGITIZATION 2 13 12 Figure 2 13 8 Old ADC Timing Diagram OLD ADC TIMING STRETCHED PULSE OF INTEGRAL SIGNAL DISCRIMINATOR LEVEL INTEGRAL SIGNAL ACQUISITION TIMING 5 or 20 microseconds 7003040A Lister Out Card The Lister Out card Figure 2 13 9 Reads the converted channel number values as they are presented on the Data Path bus If time is selected as a parameter then a time stamp is appended to each group of values The information is forwarded to the Lister card in the Workstation via the parallel connecting cable The Cytometer CPU samples the converted values over the backplane to generate the mini histograms on the Cytometer screen Provides the List Gate function Refer to Heading 5 8 in the Operators Guide
473. ring from the other direction is reduced in diameter The focal length ratio of the two lenses controls the amount of expansion reduction beam expander reducer has a focus adjustment that compensates for wavelength and allows you to set the desired amount of beam divergence Parallel Plate Consisting of a thick piece of glass a parallel plate translates a beam of light without altering the beam s direction When a laser beam enters the plate at an angle refraction bends the beam in a direction normal to the surface The beam is refracted again by the same amount when exiting the glass Figure 2 7 8 PN 4237003C 2 1 1 2 INSTRUMENT DESCRIPTION OPTICAL ELEMENTS Figure 2 7 8 Laser Beam Refraction 7003178A Light Sources Light is produced when an atom changes energy level and emits a photon of light The Elite uses a laser as a light source A laser is a useful light source because it selectively amplifies light radiation in a narrow frequency and phase range Similar to an array of antennas driven in phase by a single oscillator laser light energy is in phase and is directional High Sensitivity Flow Cell Tips As a sample passes through the laser spot fluorescent light radiates outward in all directions The mirrors radius is the same as the distance between the mirrors surface and the center of the flow channel Therefore emitted light striking the mirror reflects back toward the intersection point and toward the pick
474. rks with the PSH Peak Sense and Hold cards to perform the compensation The discriminator circuit monitors the pulse entering the PSH card This circuit also determines if a particular pulse between 0 and 10 V exceeds the discriminator level which can be set between 0 and 10 V If the discriminator level is exceeded the ADC directs the PSH cards to capture and hold the pulses that are present at that time To accomplish this the ADC controls the OPEN signal to the PSH cards this creates an acquisition window within which the system accepts the data The length of the window depends upon the type of ADC the system uses the discriminator level the acquisition start delay setting and the pulse s that trigger the discriminator Refer to the ADC and PSH Control Card under Signal Flow Description for more information 2 13 1 2 INSTRUMENT DESCRIPTION SIGNAL SELECTION AND DIGITIZATION Signal Flow Description Signal Source signals entering and Scope card have been properly amplified and processed for display acquisition and sorting by the cards described in Overview under Heading 2 13 SIGNAL SELECTION AND DIGITIZATION Mux and Scope Card The Mux and Scope card Figure 2 13 1 Selects up to eight signals for acquisition or sorting The coax cables carry the signals to the two Peak Sense and Hold PSH cards Selects two signals for pulse display A ribbon cable takes the signals to the Digiscope card for poss
475. ronic pedestal 2 Inspect the Multibus card cage to determine the type of memory card present the older 256K Memory card or the newer External Memory card a Ifan External Memory card is present 1 Remove the defective External Memory card and set it aside 2 Goto step 3 b Ifa 256K Memory card is present 1 Remove the 256K Memory card and set it aside 2 Locate and remove the CPU card 3 Install a jumper across pins 105 and 106 on the front left area of the CPU card Figure 4 13 1 Figure 4 13 1 CPU Card Jumper Locations 106 105 104 Ji _ 70032156 lt 3 Install the new External Memory card in a free slot under the CPU card Power on the Cytometer 2 Press and hold the reset switch for approximately 10 seconds until a tone is heard from the newly installed circuit card Reinstall the front left panel back on the electronic pedestal Power on the Workstation Boot to the Elite software Press 42370036 4 13 1 4 SERVICE AND REPAIR PROCEDURES EXTERNAL MEMORY CARD REPLACEMENT 10 When the prompt to transfer the program into the Cytometer appears select Yes to initiate memory transfer from the Workstation to the CPU The number 1 appears on the Cytometer right monitor and begins to sequence as the files are transferred 11 Wait while the number displayed on the right monitor sequences or counts from 1 to approximately 425 This pro
476. rs minute 2 4 gpm Laser head 27 kg 59 Ib 36 kg 78 Ib Due to low output power this laser is not recommended for sense in air operation tAir cooled laser dWater cooled laser 2 3 4 PN 4237003C PN 4237003C INSTRUMENT DESCRIPTION Table 2 3 2 Laser Technical Specifications Continued OPTICAL COMPONENTS Laser Specifications Coherent Innova 305 Wavelength output power Beam diameter at ue Beam divergence Beam pointing stability Optical noise Amplitude drift Input voltage Input current Cooling water Weight Laser head Power supply 528 7 nm 0 35 W 514 5 nm 2 00 W 501 7 nm 0 40 W 496 5 nm 0 60 W 488 0 nm 1 50 W 476 5 nm 0 60 W 472 7 nm 0 20 W 465 8 nm 0 15 W 457 9 nm 0 35 W 454 5 nm 0 12 W 363 8 nm 0 14 W 351 1 nm 0 14 W 333 6 to 363 8 nm 0 40 W 1 5mm 0 5 mrad lt 5 urads 0 2 RMS 1 208 Vac 10 50 or 60 Hz 3 phase 50 A phase 8 5 liters minute 2 2 gpm 42 kg 92 Ib 39 kg 86 Ib Coherent Spectrum Wavelength output power Beam diameter at t e Beam divergence Optical noise Input voltage Input current Cooling water Weight Laser head Power supply 647 1 nm 0 25 W 568 2 nm 0 15 W 514 5 nm 0 25 W 488 0 nm 0 25 W 476 5 nm 0 10 W 457 9 nm 0 03 W UV multiline 0 05 W 1 5 mm 647 1 nm 0 8 mrad 0 5 RMS 208 Vac 10 50 or 60 Hz 3 phase 40 A phase 8 5 liters minute 2 2 gpm 43 kg 95 Ib
477. rt tubes too low Use the sort tube holders to raise the 12 x 75 mm tubes up to the correct height within the sort area Plastic tubes were used for sorting Use glass tubes with 200 500 uL of media in the bottom Breakoff is unstable Check stability of LAD and retain stable system Less than 50 of the sort count is in the sort collection tube Low sort recovery Sort tubes are not catching all of the sort stream Usethe sort tube holders to raise the 12 x 75 mm tubes up to the correct height within the sort area 2 Increase deflection 3 Use glass tubes with 200 500 uL of media in the bottom Too few drops sorted Use a 3 drop sort window Sort delay or phase not set properly Review sort setup procedure and ensure the correct drop delay is sorted PN 4237003C 7 5 5 7 TROUBLESHOOTING TROUBLESHOOTING THE SORT SUBSYSTEM Table 7 5 1 Troubleshooting Sort Problems Continued Condition Symptom Problem Corrective Action Last attached The last attached The correct frequency has not Find the point that gives the second highest drop moves drop moves more been determined breakoff and continue with step 4 under Drive more than 3 than 3 drops Frequency Sort Settings of Heading 4 2 SORT drops when downstream to the WAVEFORM VERIFICATION AND ADJUSTMENT focusing the drops left when the frequency is adjusted for drop clarity A particular fre
478. rtion rod enters the flow cell body a If either of these conditions exist do Replacing Metal Fittings with PVC Fittings before replacing and testing the new flow cell b Ifneither of these conditions exist go to step 2 Power up the system including the laser and valves Verify that the system is at 12 0 psi Verify that the system is in vacuum Remove the new flow cell from the vial and unwrap the tissue Note The Certification Form included with the flow cell contains the crystal drive and frequency settings that gave the most stable droplet breakoff with clean crisp side PN 4237003C PN 42370036 SERVICE AND REPAIR PROCEDURES SORT SENSE FLOW CELL ADJUSTMENT AND VERIFICATION streams and at least one free drop above the ground plate In house test unit results may vary slightly Inspect the flow cell under the microscope and verify that the orifice is clean and round and in the center of the square channel that is visible behind it If the flow cell appears dirty or cloudy clean with canned air or rinse with distilled water and dry completely If the flow cell appears clean and clear go to step 7 IMPORTANT Risk of compromising fluorescence sensitivity If you touch the lens or quartz portion of the flow cell with your finger or anything else fluorescence sensitivity may be compromised Do not touch lens or quartz of flow cell 7 Screw on the new flow cell and finger tighten onto the flow body
479. rward scatter sensor assembly From the top card cage counting from the left disconnect the coax cables from the following cards and remove the cards e Slot 1 Gated Amp Control R2 Slot 6 3 PMT Sub Amp R2 Slot 7 3 PMT Sub Amp e Slot 8 Dual FL Amp Slot 49 Scat CV Amp Apply the new slot identification label over the old label on top of card cage Note Slot 1 is on the left Rest the anti static container on the top card rack Remove exiting Mux and Scope Interface card slot 6 Note The Mux and Scope card is installed in the container in slot 15 Remove the Mux and Scope Card from the container but leave it attached to facilitate the sliding of the cards into their respective slots Install the Mux and Scope card last in slot 6 of the bottom card cage Verify jumpers 1 2 and E3 E4 are in place See Figure 3 9 2 PN 4237003C INSTALLATION PROCEDURES 3 GATED AMPLIFIER UPGRADE Figure 3 9 2 Jumpers Mux and Scope Card COMPONENT SIDE 7003017A 14 Verify that PAL chip U6 PN 6704597 is installed 15 Connect the upper trace UT coax to Mux and Scope J29 16 Connect the lower trace LT coax to Mux and Scope J30 17 Connect the small blue ribbon cable from Mux and Scope J27 to Digiscope J1 18 Determine if the kit is pre wired e If the kit is pre wired install the kit and go to step 22 e Ifthe kit is not pre wired go to step 19 19 Install the new Gated Amp Control R3 card in first slot from l
480. s 8 1 4 Covers Doors 8 1 7 Cytometer 8 1 7 Filters 8 1 10 Flow Cell Area 8 1 11 Fuses 8 1 12 Hardware 8 1 12 Kits 8 1 15 Lamps 8 1 16 Lasers 8 1 16 Miscellaneous 8 1 18 Optics 8 1 19 Pneumatics 8 1 19 Power Supplies 8 1 21 Printers 8 1 21 Regulators 8 1 22 Software 8 1 22 Tools 8 1 22 Tubing 8 1 23 Workstation 8 1 23 Elite Cytometer Right Panel Removed Detail of Parts See Figure 8 2 1 8 2 1 Workstation Detail of Parts See Figure 8 2 2 8 2 2 Sheath and Waste Compartment Detail of Parts See Figure 8 2 3 8 2 3 Pneumatics Cabinet Detail of Parts See Figure 8 2 4 8 2 4 Pneumatics Drawer Detail of Parts See Figure 8 2 5 8 2 5 Pneumatics Panel Rear View Detail of Parts See Figure 8 2 6 8 2 6 Dual CRT and Connector Panel Detail of Parts See Figure 8 2 6 8 2 7 Elite Card Configuration Detail of Parts See Figure 8 2 8 8 2 8 Elite ESP Card Configuration Detail of Parts See Figure 8 2 9 8 2 9 Elite and Elite Analyzer Interconnections Detail of Parts See Figure 8 2 10 8 2 10 Multibus Card Cage All Configurations Detail of Parts See Figure 8 2 11 8 2 11 Elite ESP Data Acquisition Card Cage Detail of Parts See Figure 8 2 12 8 2 12 Cytometer Inside Rear View Detail of Parts See Figure 8 2 13 8 2 13 Cytometer Right Rear Door Inside View Detail of Parts See Figure 8 2 14 8 2 14 Cytometer Left Rear Door Inside View Detail of Parts See Figure 8 2 15 8 2 15 Cytometer L
481. s VACUUM 2 Insert the interlock defeat bar Figure 4 12 1 a Pull rod up b Open the external cover c Insert the red interlock d Push rod down to secure interlock defeat bar Figure 4 12 1 Interlock Defeat Bar EXTERNAL COVER INTERNAL INTERLOCK DEFEAT BAR 7003204A PN 4237003 4 12 1 SERVICE AND REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE 3 Remove the lens block See Figure 4 12 2 for location a Loosen the screw b Remove the beam shaper Figure 4 12 2 Location of Optical Alignment Components FLOW CELL LASER BEAM INTERLOCK VERTICAL VERTICAL DEFEAT BAR ADJUSTMENT ADJUSTMENT V ROD FLOW CELL Z AXIS ET ADJUSTMENT un im EW oo FLOW CELL O O HORIZONTAL a ADJUSTMENT HORIZONTAL ADJUSTMENT 2 SHUTTER 7003196A BEAM SHAPER SCREW 4 Open the laser shutter Note The flow cell is now rotated so the laser beam strikes perpendicular to the right side of the tip A correctly rotated flow cell directs a small amount of reflected light back through or slightly off the center of the shutter hole as shown in Figure 4 12 3 Figure 4 12 3 Laser Beam Reflected through Shutter Hole SHUTTER 7003197A 4 12 2 PN 4237003C SERVICE AND REPAIR PROCEDURES 4 OPTICAL ALIGNMENT PROCEDURE 5 Rotate the flow cell chamber on the Elite a Foran old flow ce
482. s equation PN 4237003C 2 1 3 INSTRUMENT DESCRIPTION OPTICAL ELEMENTS The Lensmakers equation is 1 1 1 BR f where f focal length n index of refraction 1 for air 1 5 for glass radius of the positive lens radius of the negative lens The more curvature a lens has smaller R the smaller the focal length f The Thin Lens equation is IO lr Ie 1 f where f focal length p distance of an object q image The Thin Lens equation relates the focal length to the distance of an object p and distance from the lens where an image q forms Magnification is determined by q p Optical Components Used in the Elite The Elite uses the following optical elements e Beam shapers Beam expanders reducers e Parallel plates e Lasers Flow cell tips Figure 2 7 5 illustrates some of the optical accessories a customer may receive depending upon the configuration of their instrument 2 1 4 4237003 4237003 JET IN AIR en amp drib OBSCURATION M FLOW GELLS FLOW CELL TIPS INSERT BAR 7003185 INSTRUMENT DESCRIPTION 2 OPTICAL ELEMENTS Figure 2 7 5 Optical Accessories DEFLECTION PLATE COVER BEAM SHAPERS CLEANING TOOLS Beam Shapers According to the refraction theory parallel rays focus to an infinitely small point at the focal point However light is a form of ra
483. s for the 100 p sort sense tip at 12 psi e 60 microseconds for the 76 sort sense tip at 12psi 7 microseconds for the 76 p jet in air tip Note With the Argon laser generated signal occurring first it is on the far left of the Digiscope screen Test this by increasing decreasing the UV laser power and observing the far right peak block the HeNe laser and observe the far right peak and raise lower the Argon laser power and observe the far left signal Verify correct alignment The Argon laser should exit near the center of the beam shaper The UV laser should exit the beam shaper slightly below the Argon They should remain parallel through the flow cell all the way to the Fals sensor Verify this by placing a business card in the beams in several different places Lower laser power is easiest on the eyes It is recommended to place the card immediately after the beam shaper just before the scatter sensor Camera Adjustment Procedure The camera assembly must be properly adjusted to obtain the best image for alignment and sorting This procedure describes how to correctly adjust the camera to achieve the following conditions Stream centered in the illuminated strobe area Strobe illumination centered on monitor Stream and strobe illumination remain centered at all zoom settings Stream to remain in focus at all zoom settings When camera position is all the way up the flow cell alignment image will remain centered on the
484. s in Sensing Area 3 7 14 Warning Labels Rear of Instrument 3 7 15 Anti Static Container of Gated Amp Kit 3 9 1 Jumpers and Scope Card 3 9 3 Gated Amplifier Assignment Screen 3 9 9 Gated Amplifier Assignment and Adjustment Potentiometer Locations 3 9 11 Gated Amplifier Control Screen 3 9 14 Gated Amp Assignment Screen 3 9 15 Standard Laser Configuration 3 9 23 Autoclone Sorting Option Mechanism 3 10 1 Laser Beam Hitting Dichroic Top View 3 12 2 Coax Cable Connections 3 13 4 Enterprise Laser Mounting Location 3 14 3 Beam Separator Location 3 14 3 Waste Bottle Caps 3 19 3 Camera Assembly Adjustment 4 1 12 Waveforms 4 2 1 Front and Back Porch 4 2 6 Pulse Signals and Histograms for Sort Coincidence Verification 4 2 14 Power Supplies 4 4 1 Fittings on Valve Assembly Bracket 4 11 2 Connecting Clip from Flow Cell to DVM 4 11 3 Filament Connecting Last Attached Drop 4 11 6 Interlock Defeat Bar 4 12 1 Location of Optical Alignment Components 4 12 2 Laser Beam Reflected through Shutter Hole 4 12 2 Flow Cell Bodies Old and Newy 4 12 3 New Flow Cell Body 4 12 4 Old Flow Cell Body 4 12 4 Parabolic Mirror and LED 4 12 5 PN 4237003C PN 42370036 4 12 8 Laser Shutter Location 4 12 5 4 12 9 Laser Beam Centered on Forward Scatter Detector Mask 4 12 6 4 12 10Scope Screen Selections 4 12 6 4 12 11 Laser Beam Horizontal Adjustment Location 4 12 7 4 12 12Laser Beam Centered
485. s not return to the position that it was before a CLEAR or DEBUBBLE was done there may be a fluidics problem Set the cursors to calculate the delay a Zoom the camera out to get the full pan function to reach the laser intersection point and the first free droplet b Place the cursors Refer to the Operators manual for instructions Verify the flow factor adjustment by performing a matrix and determining the correct delay at the Sort screen Do Delay Calculation under Heading 4 2 SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE If you were doing Replacing Metal Fittings with PVC Fittings resume that procedure at step j 4 11 7 4 SERVICE AND REPAIR PROCEDURES SORT SENSE FLOW CELL ADJUSTMENT AND VERIFICATION 4 11 8 PN 4237003C SERVICE AND REPAIR PROCEDURES 4 OPTICAL ALIGNMENT PROCEDURE 4 12 OPTICAL ALIGNMENT PROCEDURE Purpose Use this procedure to align the flow cell and the laser beam The procedure is divided into three sections Flow Cell Alignment Laser Beam Alignment and Optical Fine Tuning Tools Supplies Needed None Flow Cell Alignment WARNING Risk of personal injury The laser beam can cause eye damage if viewed either directly or indirectly from reflective surfaces such as a mirror or shiny metal surface To prevent eye damage avoid direct exposure to the beam Do not view it directly or with optical instruments except with special service tools as directed in this manual 1 Pres
486. screw PN 4237003C SERVICE AND REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE Positioning Movement of the Camera When camera position is all the way up the flow cell alignment image must remain centered on the monitor at all zoom settings If the image does not remain centered do this procedure See Figure 4 1 1 1 Move the camera up until it stops ATTENTION Do not lose the setscrew Do not allow the belt to fall off 2 Loosen the screws holding the position pot pulley and move do not rotate the pulley to release tension on the belt 3 Rotate the pulley slightly to move the camera to a new position and stop the movement by returning the pulley to its original position Repeat this step until the image is correct 4 Move the pulley to tighten the belt and tighten the screws Optimizing the Image Adjust the camera video level for best image definition as follows Remove the left front panel from electronics pedestal Use the camera to observe sheath flow Adjust R99 on the Camera Interface card to obtain the best camera image Note This control is a video gain adjust that controls the brightness of the image Increase the brightness until the image begins to bloom or saturate Decrease the brightness until good image quality is observed PN 4237003 4 1 11 4 SERVICE AND REPAIR PROCEDURES OPTICAL ALIGNMENT PROCEDURE 4 1 12 Figure 4 1 1 Camera Assembly Adjustment Z00M MECHANISM Z00M POT PULLEY
487. se present d Change the delay to 7 microseconds and the clock to 0 5 At the Gated Amp Assign screen set AUX2 to the LWBM At the Scope screen a PutPARM Bon the top trace b Adjust R138 slot 11 to minimize At the Gated Amp Assign screen a Select AUX2 to UPBM b Adjust R119 slot 9 to minimize At the Gated Amp Control screen a Change the delay to 60 microseconds and the clock to 5 b Adjust R110 slot 10 to minimize PN 4237003C PN 42370036 34 35 36 37 38 39 40 41 42 43 44 45 46 47 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE c Change the delay to 7 microseconds and the clock to 0 5 d Change the trigger source to PMT1 PEAK e Verify that pulses are going into PMT 1 and arrange optical filters f Check Trig Signal of PMTI Peak At the Scope screen put PMT2 LOG on the upper trace At the Gated Amp Assignment screen a Ensure PMT2 is on the LWBM b Adjust R197 slot 6 to minimize c Select PMT2 UPBM d Adjust R106 slot 8 to minimize At the Gated Amp Control screen a Change the delay to 60 microseconds and the clock to 5 b Adjust R107 slot 7 to minimize c Change the delay to 7 microseconds move the blank to the bandpass position under PMT3 and change the clock to 0 5 At the Scope screen put PMT3 LOG on the upper trace At the Gated Amp Assign screen a Ensure is on the LWBM b Adjust R198 slot 6 to minimize
488. set pots to adjust for offsets in the 7 Microsecond Delay card Do this for PMT1 PMT2 PMT4 PMT5 installed AUX1 and AUX2 Note When setting the PMT to be tested for 1 division tall log on trace 2 on the scope disable the gated amp Note Use a sensor not being tested for the gated amp trigger source so amplitude will be large enough to start gating process At the Gated Amp Control screen select the following PMT1 PEAK TRIGGER SOURCE CLOCK 5msec WINDOW WIDTH 20 msec WINDOW DELAY 50 msec SIGNAL DELAY 60 msec Note WINDOW DELAY can be adjusted as necessary 30 45 50 etc to capture complete pulse Repeat step 50 for PMT1 PMT2 PMT3 PMT4 PMT5 AUXI amp AUX2 for 60 microsecond pulse delay using Table 3 9 4 for pot for signals needing adjustment Adjust the offsets for the 7 microseconds delay using the same procedure in step 50 The pots will be different for each card See Table 3 9 4 Do the correct laser alignment Laser Alignment With Gated Amplifier Laser Configuration One Innova 305 laser is set to the UV mode and placed in the rear most position on the optical bench and parallel to the PMT block A beam expander contractor is used to insure beam diameter is the same for both lasers A second Innova 305 laser is set to the 457 nm line and placed parallel to the first Innova 305 laser If necessary it is possible to position these two lasers close enough together so allow both the Arg
489. sheath and fix 2 Replace the regulator Fanning recurs shortly after clear or debubble temporarily fixes the problem Pinched sheath tubing Follow sheath tubing from the flow body to release the constriction Note Tugging gently on the sheath tubing at the point before it disappears behind the sort collection area may alleviate the problem 7 5 2 42370036 Table 7 5 1 Troubleshooting Sort Problems Continued TROUBLESHOOTING TROUBLESHOOTING THE SORT SUBSYSTEM Condition Symptom Problem Corrective Action Fanning There is a Insertion rod is corroded Fix this problem before continuing the sort between side blackish greenish causing an irregular droplet and center buildup on the charge and flow cell plugs insertion rod in the Flow through fittings on the Find cracked or damaged fitting and clean entire continued flow body valve bracket assembly are wrong metal or one of the correct flow through fittings PVC is cracked system of the metal fillings created from the electrolysis of the rod as instructed under Heading 4 11 SORT SENSE FLOW CELL ADJUSTMENT AND VERIFICATION Fanning occurs after replacement or adjustment of flow cell Loose flow body Loose flow cell tip Ensure the flow cell and the flow body are sufficiently tightened Loose alligator clip Oil the hinge on the alligator clip Bimorph is unstable Replace the bimorph usin
490. sion Card 1 At the electronics pedestal remove the right panel to gain access to the data acquisition electronics a Remove the four Phillips head screws securing the panel b Disconnect the grounding cable from the Cytometer chassis C Setthe panel aside 2 Locate the Bitmap and Sort Decision card in slot 12 of the lower card cage a Remove the ribbon cable attached to the card b Remove the circuit card C Set it aside 3 Install the new Bitmap and Sort Decision R2 card in the slot in slot 12 Reconnect the ribbon cable so that the arrow on the connector is pointing up when the cable is attached 5 Reinstall the right panel on the electronic pedestal a Reconnect the grounding cable from the Cytometer chassis b Secure the panel using the four Phillips head screws removed earlier Install The Elite Software l Locate the Elite version 4 5 Software disk 1 of 2 2 Insert disk 1 into the 3 5 inch floppy drive Note If the Workstation has two floppy drives the 3 5 inch drive is designated as the B drive If the 3 5 inch drive is the only floppy drive the drive is the A drive 3 Ifthe Workstation is presently in the Elite Software press then to exit to DOS At the CAELITE prompt type A then press Enter To access the README document Ifthe Cytometer does not have a printer go to step 6 Ifthe Cytometer is equipped with a printer go to step 7 PN 4237003 3 22 1 INSTALLATION PROCEDURES ELIT
491. sion Card Bitmap to DATAPATH From prism sort download window test card from dt interface card BITMAP DOWNLOAD LOGIC Compare window results to sort CRITERIA criteria to determine whether to lt sort based on linear regions BITMAP 1 Capture parameter pairs and address BITMAP 2 location to test in BITMAP 3 appropriate bitmap BITMAP 4 DNS DNS BITMAP RESULTS LEFT RIGHT BITMAP Compare bitmap results CRITERIA to stored bitmap criteria lt MEMORY to determine whether to sort based on bitmap 45V 7 S STROBE DO NOT SORT DNS LEFT LEFT DO NOT SORT DNS RIGHT SORT RIGHT 7003054 CLOCK OUTPUT In Figure 2 8 4 the bitmap download logic decodes the incoming data word to address and set a particular location in a bitmap Interface and Scaler Card The Interface and Scaler card e Provides digital counters for various sort actions sort right sort left abort counts right and left e Compares the sort counts to the sort stop values When the card reaches the stop value it signals the Sort Oscillator card to stop the sort e Records other system events via counters including acquisition elapsed time and acquisition stop count 4237003 2 8 7 2 INSTRUMENT DESCRIPTION SORT SUBSYSTEM 2 8 8 Sort Delay Card The Sort Delay card Figure 2 8 5 Delays the application of the sort pulse until the cel
492. solenoid valves Autoclone Sorting Option drain function of 2 9 2 Autoclone Sorting Option waste function of 2 9 2 bubble drain function of 2 9 2 drip chamber drain function of 2 9 2 flow cell vacuum function of 2 9 2 laser shutter function of 2 9 2 operation illustration of 2 9 3 Pneumatic Interface card function of 2 9 4 rinse flow function of 2 9 2 rinse pressure function of 2 9 2 sample collect drain function of 2 9 2 sample pinch function of 2 9 2 sample pressure function of 2 9 2 sheath pressure function of 2 9 2 system pressure function of 2 9 2 VLI function 2 9 2 VL2 function 2 9 2 VL3 function 2 9 2 VL4 function 2 9 2 VL5 function 2 9 2 VL6 function 2 9 2 VL 7 function 2 9 2 VL8 function 2 9 2 VL9 function 2 9 2 VL10 function 2 9 2 PN 4237003C INDEX VL11 function 2 9 2 VL12 function 2 9 2 VL13 function 2 9 2 VL14 function 2 9 2 VL15 function 2 9 2 water drain function of 2 9 2 solenoids See solenoid valves sort coincidence verification procedure 4 2 9 sort current leakage test 7 5 1 7 6 1 Sort Delay card block diagram 2 8 9 f20 clock function of 2 8 8 function 2 8 8 verifying sort coincidence after replacing the card 4 2 9 Sort Delay R3 card jumpers 4 10 3 A 2 3 sort function adjustments based on direction beads fell 3 6 6 applications list of 2 7 6 cable connections required 3 13 4 checking the side streams 3 6 5 circuit card requir
493. sort gate on a fluorescence signal Verify that the droplets sorted is set to 1 Cv Scb Set the Sort Stop to 100 to make it easy to see the correct drop quickly in the fluorescent microscope 7 Runasortseven times once each for Delay settings from three less to three more than the values given on the Camera screen 8 View the sorted drops under the fluorescent microscope beads should be all in one drop about 9096 of the beads in one drop is fine If so select the drop and go to step 13 e fall 29096 the beads are not in one drop select the drop containing most of the beads For example for purposes of illustration assume most of the beads are in drop 29 and some are in 30 In this case you would select drop 29 9 View the selected drop on the monitor and place a cursor tangent to the tip of it Note Verify that the camera view is zoomed in sufficiently to easily view the last drop and the filament connecting it to the stream 10 To make the beads fall in the selected drop 29 in our example instead of 30 raise the flow cell very slightly by turning the flow cell height adjustment to raise the flow cell and stream 1 16 of a drop while watching the camera 4 2 7 4 SERVICE AND REPAIR PROCEDURES SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE 11 12 13 14 15 16 Note Make sure that the fluorescence signals do not drop In quartz this adjustment is more sensitive than i
494. stments to obtain linear operation over the whole operating range PN 4237003C 2 9 5 INSTRUMENT DESCRIPTION PNEUMATICS SUBSYSTEM 2 9 6 Figure 2 9 2 Block Diagram Sample Pressure System Old Type Regarding Figure 2 9 2 1 gt 8 00 V 1 Tre 5 9 P 1 VOLT Y VOLT VOLT REF REF REF 1 R31 R34 1 1 REF SENSOR OUT VOLT R59 TP7 l e R51 SENSOR Y INPUT LATCH DAC LU 1 1 CPU v Y 1 1 FEEDBACK OP AMP DRIVER l DRIVE 1 R63 SAMPLE REGULATOR 8 15 PSI 0 10 VIN 7003074A 30 PSI IN The Input Latch is an 8 bit word set pressure written by CPU U23 The DAC provides a voltage proportional to the digital pressure Full scale is set by Volt Ref PN 4237003C PN 42370036 New INSTRUMENT DESCRIPTION PNEUMATICS SUBSYSTEM The new type of Sample Pressure System Figure 2 9 3 consists of a DAC that converts the requested pressure in digital form to a 0 10 V analog level This level goes to the low bleed sample regulator The regulator uses an internal pressure transducer to sense and regulate the sample pressure separate circuit consists of a pressure transducer a reference power supply and an amplifier to provide a voltage
495. system pressure regulator until the system pressure is 30 psi Slowly reduce system pressure on the Compressor module until the DVM reads 4 to 5 V The pressure displayed on the system pressure gauge should be 17 psi If not adjust the screw on the system pressure switch and repeat steps 7 and 8 until the correct value is obtained 4 7 1 4 SERVICE AND REPAIR PROCEDURES PRESSURE SWITCH ADJUSTMENT 4 7 2 PN 4237003C 4 8 PN 42370036 WORKSTATION AND SOFTWARE PC Model 486 Setup Procedure At the Workstation press Cti Alt Delete to reboot the Workstation pc During the reboot sequence press to display the Setup menu qe Eb Select STANDARD CMOS SETUP SERVICE AND REPAIR PROCEDURES WORKSTATION AND SOFTWARE Follow the instructions on the screen and ensure the following configuration Date mm date year current date Time hour min sec current time Hard Disk C Type 1 Hard Disk D Type Not Installed Floppy Drive A 1 2 MB 5 1 4 Floppy Drive B 1 44 MB 3 1 2 Primary Display VGA PGA EGA Keyboard Installed Base memory 640 KB Extended Memory 15360 KB Cyln Head WPcom LZone Sect 306 4 128 305 17 Press to return to the Setup menu Select ADVANCED CMOS SETUP Size 10 MB Follow the instructions on the screen and ensure the following configuration ADVANCE CMOS SETUP Typematic Rate Programming Disabled Typematic Rate Delay msec 500 Typematic Rate chars Sec
496. t Note It is recommended that you start at 32 kHz and scan down 4 When the highest breakoff is achieved and the droplets appear very focused and sharp stop scanning Note Highest breakoff means closest to the flow cell 5 Seta cursor tangent to the bottom of the last attached droplet Note If the last attached droplet is not yet visible above the top of the ground plate increase the crystal drive from 75 to 85 If the droplet is still not visible move the deflection body slightly down Turn OFF the High Voltage Press DEBUBBLE twice and verify that the breakoff returns to the same place If the breakoff does not return to the same place the flow cell has a bubble or is partially clogged Remedy this situation before continuing Note You may have to remove or replace the flow cell 8 Move the camera all the way down stream and look at the droplets The droplets should be clear sharp and fairly round in shape there should be no satellite droplets If there are satellite droplets then proceed to step 9 are no satellite droplets slightly adjust the frequency via the decimal setting to improve the shape and clarity of the droplets 9 Move the camera back upstream and ensure that the last attached droplet position has not changed by more than three droplets You may have to adjust the sort deflection body in order to view the last attached drop tis helpful if the last attached droplet
497. t hex 47 32 UNS x 0 562 Analyzer 8 2 18 2 2826051 Washer split lock 0 26 i d x 0 49 o d Argon laser 8 2 34 4 2830014 Grommet sample tube 0 375 i d x 0 625 o d x 0 250 8 2 1 20 2839039 Screw self locking 6 32 x 0 37 analyzer 8 2 18 20 24 2839043 Screw self locking 6 32 x 0 62 analyzer 8 2 18 16 2839058 Screw self locking 4 40 x 0 18 analyzer 8 2 17 6 2851558 Screw machine 25 20 x 0 62 Elite Analyzer 8 2 20 3 11 2851836 Screw thumb 6 32 x 0 25 analyzer 8 2 20 13 2851876 Thumbscrew RPL analyzer sample 8 2 17 3814206 Screw metric micro optical rail 6011002 Tie wire nylon 0 14 x 6 7 8 2 35 7 6211015 Valve actuator analyzer 7 16 bore 8 2 5 8 8 2 18 14 23 8 2 19 1 6216012 Spacer cylinder 0 500 i d x 0 562 o d x 0 062 8 2 18 13 6216129 Fitting hose barb T brass 0 187 i d 8 2 6 10 PN 4237003C 8 1 13 8 PARTS LISTS MASTER PARTS LIST IN NUMERICAL ORDER BY CATEGORY Table 8 1 10 Hardware Continued Part Number Description Figure Item 6216345 Gasket 10 black 8 2 18 10 6232086 Fitting hose barb 0 062 i d to 10 32 thread 8 2 18 17 6232092 Fitting quick connect white female 8 2 3 2 8 2 4 8 6232093 Fitting quick connect white male 6232104 Fitting hose barb T 0 115 to 0 180 i d 8 2 6 5 6232124 Fitting hose miniature 10 32 analyzer 8 2 18 9 6232205 Fitting barb elbow nylon white 0 125 i d to 10 32 8 2 17 11 8 2 18 7 6232208 Fitting hose barb elb
498. t 13 J11 3 PMT Sub SW R3 slot 12 Scat Aux Gated Amp slot 11 J19 Scat Aux Gated Amp slot 11 J20 3 PMT Sub SW R3 slot 12 J2 3 PMT Sub SW R3 slot 12 J3 3 PMT Sub SW R3 slot 12 J5 3 PMT Sub SW R3 slot 12 J6 Scat Aux Gated Amp slot 11 J21 J J J Peak Scatter Mux slot 13 J8 Peak Scatter Mux slot 13 J6 3 PMT Sub SW R3 slot 12 J7 3 PMT Sub SW R3 slot 12 J8 Peak Scatter Mux slot 13 J1 J J J Peak Scatter Mux slot 13 J5 3 PMT Sub SW R3 slot 12 J4 Scat CV SW R1 slot 14 J1 Peak Scatter Mux 27 in cable slot 13 J2 Scat CV slot 14 J6 Peak Scatter Mux 27 in cable slot 13 J3 Scat CV slot 14 J5 3 PMT Sub SW R1 slot 5 J6 PMT Gated Amp 27 in cable slot 6 J2 Scat Aux Gated Amp slot 11 J4 Peak Scatter Mux 27 in cable slot 13 J9 PMT Gated Amp 27 in cable slot 6 J3 Scat Aux Gated Amp slot 11 J5 Scat CV 27 in cable slot 14 J8 Dual FL slot 3 J6 Scat CV 27 in cable slot 14 J10 Peak Scatter Mux slot 13 J12 Gated Amp Control R3 27 in cable slot 1 J3 Scat Aux Gated Amp slot 11 J1 Gated Amp Control R3 27 in cable slot 1 J4 PMT Gated Amp slot 6 J1 Mux and Scope J29 Digiscope LT Mux and Scope J30 Digiscope UT Mux and Scope J26 No Connection Mux and Scope 27 in cable J9 Peak Scatte
499. t TP4 Set Sheath Pressure to 3 00 psi After the pressure stabilizes change the set pressure slightly as needed for the gauge to read 3 00 psi Adjust R76 for 2 00 0 003 Vdc at TP4 Repeat steps 11 through 14 until no further adjustment of R75 or R76 is needed Remove the jumper from E2 to and place on El to E2 Set Sheath Pressure to 12 00 psi on the Cytometer Control screen Adjust R81 for 12 0 0 02 psi on the gauge Set Sheath Pressure to the following pressures and verify that the set read and gauge pressures agree within 0 1 psi 11 5 psi 4 5 psi Repeat steps 13 through 20 if needed Set Sheath Pressure to 5 00 psi Verify that the set read and gauge pressures agree within 0 2 psi Repeat steps 13 through 20 if needed Remove the gauge Low Bleed Regulator Adjustment These regulators do not normally require adjustment However if difficulty is encountered with pneumatic adjustment the regulators can be tested and adjusted as described in this procedure 4 5 2 42370036 PN 42370036 SERVICE AND REPAIR PROCEDURES PNEUMATICS SYSTEM IMPORTANT Risk of erroneous results if the sample and sheath regulators are adjusted using noncalibrated test equipment Use only calibrated test equipment to perform the following procedures Sample Regulator Musee d D 9 10 11 Connect the to TP8 Connect the external gauge to the sample pressure line Set the Sample Pressure to 15
500. t mode where the feedback from the liquid pressure transducer is disabled and the pressure regulator uses its internal air pressure sensor for feedback In the test mode the Pneumatic Control card does not compensate for sheath liquid level changes which simplifies the card s adjustment and troubleshooting 2 9 10 PN 4237003 INSTRUMENT DESCRIPTION 2 PNEUMATICS SUBSYSTEM Figure 2 9 5 Block Diagram Sheath Pressure System New Type 1 TP10 R81 10 00 V VOLT 1 10 00 V R54 TP3 VOLT I EC REF gt am INPUT LATCH DAC 11 l 1 4 NM 1 R75 GAIN lt R76 ZERO 1 1 O J4 TO SENSOR 1 0 1 NZ E3 E E1 to E2 normal operation sheath sensor feedback 2 to adjustment setting regulator uses internal sensor only 1 MEM ME EE secondary SHEATH feedback REGULATOR input SENSOR LIQUID Jolt E TRANSDUCER AMP amp 3 15 PSI 15 DRIVER OUT Vv 30 PSI IN 7003077A Regarding Figure 2 9 5 The Input Latch is an 8 bit word set pressure written by CPU U17 The DAC provides a voltage pr
501. t parameter is greater than 9 8 V or channel 1 000 Figure 2 8 9 Interconnection of Pulse Pileup Det TOF Card SLOT 5 PULSE PILE UP amp TIME OF FLIGHT COAX INTERCONNECT 10 TURN PPU TOF BMWDTH NORM QPSH 1 J2 SLOT 7 CH 1 OUT 2 _ gt DIAL CH 8 OUT J4 i 3 PANEL MOUN f pe SEN CH B J5 STRETCH JUMPER PLS IN EE J6 II PPU TOF CARD PARAM 8 OU dies JUMPER J7 LOT 8 ST E1 TO E2 Ser LEGEND E20 TO E21 CH 8 IN CH 8 m CH 8 USE 3 17 IN IN STRETCH J3 P PLS OUT 60270793 CR AE 9 93 JUMPER 2 4 J EXISTING T CH 8 CH 1 ig SLOT 9 17 OUT OUT ER IA LAN PARAM 8 MUX amp SCOPE SITE ADC PSH CONTROL 7003043A PN 4237003C 2 8 15 INSTRUMENT DESCRIPTION SORT SUBSYSTEM TOF Function The TOF function Figure 2 8 10 of the Pulse Pileup Det TOF card measures the length of the signal assigned to parameter eight Figure 2 8 10 TOF Function of Pulse Pileup Det TOF Card MUX ADC AND AD AND RUA PSH CONTROL SCOPE x CARD J8 J7 J8 J8 CHAN 8 CHAN 8 o 8 OUT IN IN J3 J4 J5
502. t to switch to the PPU Det mode Note that a new selection block labeled PPU sensitivity now appears This block switches among four states e 0 5 to 4 microseconds 4to 10 microseconds 4 2 11 4 SERVICE AND REPAIR PROCEDURES SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE 4 2 12 10 to 20 microseconds Over 20 microseconds Note Choose the range that encompasses the pulse width of peak pulse being used for PPU In this test the 10 to 20 microsecond range should be used In normal operation the range will depend on the flow cell e 76 p jet in air 0 5 4 e 76 psort sense 10 20 e lOO0nsortsense 4 10 The sort counts stop and the abort rate increases this verifies operation of the PPU circuit Verification of Sort Pulse Extension The sort pulse is lengthened if a second good cell appears in the droplet immediately following the sort envelope for the first cell EE P WE D ON D Turn Coincidence to Coinc Abort OFF Set Drops Sorted to 1 Adjust R109 for 80 microsecond delay Observe the sort pulse on the oscilloscope Note that the pulse consists of a front and back porch but no center level Slowly adjust R109 to decrease the pulse delay When the pulses are approximately 60 to 30 microseconds apart the center level for the sort pulse appear this indicates the one drop sort is now being extended into a two drop sort Adjust R109 so the two pulses are almost touching Change Drops Sorted to 3 Observe th
503. te line 8 2 6 2 6855931 Lines air and vacuum 6856539 Drawer pneumatics 6856634 Box gauge and regulator pneumatic housing 6856677 Compressor module 115V 6856699 Pneumatics cabinet 8 2 4 N A 8 1 20 42370036 MASTER PARTS LIST IN NUMERICAL ORDER CATEGORY Table 8 1 16 Pneumatics Continued PARTS LISTS Part Number Description Figure Item 6856859 Table top laser mount 8 2 4 1 6856889 Door pneumatic side 8 2 4 5 6856984 Chamber lower bubble 8 2 6 3 6856985 Bubble chamber upper 8 2 6 1 6857576 Valve pinch double acting 8 2 5 9 6857872 Rinse tank assembly 8 2 4 2 Table 8 1 17 Power Supplies Part Number Description Figure Item 2603058 Fan blower 90 CFM 24 Vdc 4 69 sq in 1 in thick 4004024 x90 Vdc 0 2A 115 Vac 8 2 13 14 4004074 Switcher 400 W 8 2 13 8 4004084 Power conerter 1 3 K Vdc out 5 12 Vdc in 6704960 Deflection 8 2 13 15 6705493 3000 V Deflection 8 2 15 2 6804637 PMT HV Bertran 8 2 13 4 6853275 15 2 0 8 2 13 10 6854358 15 V 5 0 A 8 2 13 12 6856870 15 V 0 25 A 8 2 13 11 6856636 PMT HV 8 2 13 2 6857017 HeNe laser 6857226 Fiber optics 7260017 Computer 220 W 8 2 28 2 Table 8 1 18 Printers Part Number Description Figure Item 2016675 2016691 2016692 2016693 2016694 4237252 6027510 HP Deskjet 1600C Color Printer Cartridge ink HP 1600 Color Printer magenta Cartridge ink HP 1600C Color Printer black Cartridge ink HP
504. teps through each well on the plate and 3 10 8 returns to the Load position when complete Note The wells turn red since no sort pulses are present PN 4237003C PN 42370036 INSTALLATION PROCEDURES AUTOCLONE SORTING OPTION INSTALLATION Second Test Set FL Test DRIVE to 100 Start acquisition and ensure a population builds up within the sort region Test created earlier Select Activate and observe that the Autoclone Sorting Option steps through the wells of the plate Note The wells turn green indicating a successful sort for each well Stream and Waste Catcher Adjustment When sorting with the Autoclone Sorting Option the Deflection Gain and the Stream control located above the fluidic controls are adjusted such that the waste stream is constantly deflected to the right into the waste catcher and the sort steam drops straight down on the collection plate The waste catcher has two positions In the extended position it catches the waste stream and the sort stream When retracted it catches only the waste stream 1 Turn on Sheath and use Sort Test to setup sort streams Verify stable droplets and sidestreams Turn the Stream knob until the left sort stream is flowing straight down Switch the deflection high voltage on and off to check the stream position With high voltage on the left stream should look the same as the sheath stream does when high voltage is off Note You can control the waste catche
505. the trigger Compares the selected trigger signal to a programmed dc level trigger level and activates the gate window generator if the trigger level is exceeded The generator is based on two timing circuits The first provides a time delay starting from when the trigger level is exceeded The time delay is the window delay period After the window delay period ends a second timing circuit generates a programmed length pulse which is the window width pulse When enabled this pulse is routed to the Gated Amp card which then gates the actual signals Selects the window pulse gate window or the trigger source to send to the Mux and Scope card for display Intercepts the command to select which channels are to be delayed This command is decoded and used to toggle individual lines on the backplane that go to the Gated Amp card s which actually selects the delays Prevents retriggering of the timing circuits until the current timing cycle is completed when the Lower Beam Lockout is selected When the Gate Window Discriminator Override is selected the gate window is routed to the Mux and Scope card where it is connected to the DISCSAT line forcing the Gated Amp Discriminator Level to become the System Discriminator Scat Aux Gated Amp Card Functions The Scat Aux Gated Amp card Figure 2 12 2 Selects one of the four signals to be the trigger source and routes the signal to the Gated Amp Control card Sends all four signals to th
506. the ac power outlets Note When forward scatter FS is blocked no signal should be visible If offset is wrong you will see a signal either before the window offset or after the window offset Adjust R135 slot 11 Scat Aux Gated Amp to minimize any pulse present on the upper trace during or after the window View the scope output to see zero crossing Select FALS UPBM and adjust R106 slot 9 to minimize any pulse present on the upper trace during or after the window Repeat steps 13 through 14 with signals set one decade higher and center the adjustment potentiometer At the Gated Amp Control screen Figure 3 9 5 a Change the delay to 60 microseconds and change the clock to 5 b Adjust R107 slot 10 to minimize any pulse present on the upper trace during or after the window c Turn Strobe off At the Options screen select Align LED Pulsed 100 and Drive 2 kHz Replace the mirror with the 488 DL filter Select Display Trigger a Change trigger source to PMT4 PEAK b Setthe delay to 7 microseconds and change the clock to 0 5 c Adjust PMT 4 HV for 3 divisions tall trigger pulse Note If PMT 5 is installed change trigger source to 5 PEAK Set the delay to 7 microseconds and adjust PMT5 HV for 3 divisions tall TRIGGER At the Scope screen put 1 LOG on the top pulse 3 9 19 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE 3 9 20 23 24 25 26 27 28 29 30
507. the other end of the cable through the side of the computer pedestal and connect it to the Lister card connection on the back of the Workstations CPU 3 4 2 PN 4237003 INSTALLATION PROCEDURES 3 CONNECTING ASSEMBLIES 7T Connect the serial comm cable a Connect one end of the cable to the Cytometers SERIAL COMM connector as shown in Figure 3 4 2 b Feed the other end of the cable through the side of the computer pedestal and connect it to the COM2 port on the Workstations CPU Plug the Workstation power cord into one of the Cytometers ac receptacles Plug the color monitors power cord into the Cytometer s remaining ac receptacle 10 Carefully press all the cables going to the Cytometer into the wiring channel under the table top Tubing Connections l Prepare the rinse bottle a the rinse bottle with deionized water and place the bottle in its holder b Attach the quick connect tubing connectors to the appropriate connector Be sure to match the colors c Plug the level sense connector into the matching socket 2 Prepare the sheath bottle a Fill the sheath bottle with conductive sheath fluid such as IsoFlow sheath fluid and place the bottle in its holder See Figure 3 4 3 for location b Attach the quick connect tubing connectors to the appropriate connector Be sure to match the colors c Plug the level sense connector into the matching socket 3 Prepare the waste bottle a Attach the qu
508. the waste level sensor logic to provide fail safe monitoring of the level sensing circuits The software does not lock up when Acquisition is started and Sort Logic is changed on the Cytometer Sort screen from Ext Sort to Cmp Abort and Acquisition is stopped user then goes to Sort screen at the Workstation The software does not lock up when the user prints a bar code label for a Panelyzer run under the Acquisition application then switches to a different application then back to Acquisition The software reads FCS 2 0 listmode and histogram files The software reads large listmode file 210 MB During data analysis statistics display all asterisks when an overflow in a channel occurs 265535 events in 1 channel or when an overflow is a histogram occurs The software allows the user to switch PPU sensitivity ranges independently from sort logic This procedure applies to Elite Flow Cytometer 2362 Elite Analyzer Flow Cytometer 2819 Elite Flow Cytometer with Gated Amp and or Autoclone Sorting Option 2360 Elite ESP Flow Cytometer 2358 Elite ESP Flow Cytometer with Gated Amp and or Autoclone Sorting Option 2357 Elite ESP Flow Cytometer with Gated Amp and or Autoclone Sorting Option and Advanced Lasers 2356 High SN W48072 Low SN V39054 Version 4 01 detects the current version of software installed on the instrument and if version 4 0 is not detected you cannot install version 4 01 Version 4 01 disks contain an updated
509. through a parallel port The solenoids are controlled through this pathway Controls the remaining cards in the Cytometer through the Data Taker Interface card See Figure 2 5 3 for details on control pathways External Memory or 256K Memory Card The External Memory or 256K Memory card provides additional memory RAM for the Cytometer program to run When the system is powered off or when the External Memory or 256K Memory card is removed from the unit the RAM is powered by lithium batteries The RAM draws 250 microamps of current from the batteries At this rate the life of the batteries is the same as their shelf life five years Serial 1 0 Card The Serial I O card is a five channel RS232 interface card that allows The CPU to communicate with the KIR3 touch screen card and the Indicator card the Sample Station control panel through one channel For communication with the Workstation through another channel Laser Control Subsystem The Laser Control subsystem Figure 2 5 1 consists of the Dual Laser Control card Laser relays located in the transformer drawer Interlock switches Relay panel on systems equipped for additional lasers Interlock keyswitches Dual Laser Control Card The Dual Laser Control card has two main functions 2 5 2 The interlock function which allows lasers to turn on only if the interlock switches are correctly configured this function does not involve the CPU card Laser control
510. through the upper beam the PI dye emits light according to how much is present in the cell As the cell passes through the lower beam the Hoechst dye emits light according to the concentration of the dye in the cell A delay line retards the signal from the upper beam by a time equal to the time required for the cell to fall from the upper to the lower beam This occurs because the acquisition electronics expect all signal for a particular cell to be synchronous The delay is 7 microseconds for a sense in air flow cell This corresponds to a beam separation of 70 p because the sheath flow outside the aperture is 10 meters second The flow rate inside the Sort Sense flow cell is much less than that outside the aperture Therefore the beam separation determines which delay line is used the 20 40 or 60 microseconds The Gated Amp Detects the cells by comparing the selected trigger source signal usually FALS to a trigger discriminator level Delays all signals generated by the upper beam Allows the delayed and non delayed signals to pass on to the amplifier cards as the cell passes through the lower beam Refer to the individual card descriptions for information on where and how the Gated Amp functions are implemented Signal Flow for Each Configuration Table 2 6 2 summarizes the Elite configurations regarding signal flow Table 2 6 2 Configuration Summary Generation 4 PMT Nonswitchable Amps 5 Nonsw
511. tional test See VIP pressure regulator adjustment 4 6 1 safety precautions to follow 1 2 1 system description of 2 4 1 system See system electronics electrostatic buildup preventing See Autoclone Sorting Option Elite 256 Sort Upgrade installation procedure 3 22 1 Elite manuals list of 1 1 1 emission spectra See color compensation engineering schematics included in this manual 6 1 1 equations used by instrument Lensmaker s equation 2 7 4 log amplification 2 6 3 Snell s Law 2 7 2 Thin Lens equation 2 7 4 error messages Drip Chamber Error 2 9 1 Waste Filter Error 2 9 1 ESP installation See sorting ESP instruments summary of circuit card upgrades 3 13 5 upgrade procedure 3 13 1 event defined 2 13 1 extended abort See sort function verification External Memory card PN 4237003C INDEX upgrade procedure 4 13 1 F f20 clock function of 2 8 8 FALS adjustment procedure 3 9 10 fans cooling fans illustration of 3 5 1 locations illustration of 3 5 1 feedback system closed loop 2 9 5 fiber optics power supply adjustments and measurements 4 4 2 A 3 2 FIFO register function 2 13 14 filament connecting last drop See sorting file paths B 1 1 filters waste function of 2 9 1 water function of 2 9 1 filters optical See optical filters fittings defective locating 4 11 3 replacing on valve bracket assembly 4 11 1 FLI signal flow summary of 2 6 9 FL2 signal flow summary of 2 6 9 FL3 signa
512. tive maintenance See PMI procedure principles circuit operation 2 6 4 Printers HP 1200C See HP 1200C color printer HP 1600C color printer See HP 1600C color printer initial installation connections for 3 4 4 Prism and Sort Window Test card block diagram 2 8 5 digital values reading 2 13 1 function 2 8 4 jumpers 4 10 3 A 2 3 protocols creating for acquisition check 3 6 1 PSH cards See Quad PSH card pulse pileup verification See sort function stretched defined 2 13 4 width variations of 2 6 5 pulse generator adjustments See sort function verification Pulse Generator and Clock R card jumpers 4 10 3 A 2 3 Pulse Generator card adjustments for sort pulse truncation test 4 2 12 connections for pulse generator setup 4 2 10 jumpers 4 2 14 4 2 15 potentiometer functions 4 2 15 test point functions 4 2 15 pulse generator setup See sort function verification pulse heights obtaining maximum 4 12 12 Pulse Pileup Det TOF card function 2 8 15 interconnection illustration 2 8 15 jumpers 4 10 3 A 2 3 PPU function 2 8 17 PPU function illustration of 2 8 18 TOF function 2 8 16 TOF function illustration of 2 8 16 TOF verification 5 1 2 verifying sort coincidence after replacing the card 4 2 9 Pulse Pileup Det TOF card verification procedure 4 2 11 pulse video display components of 2 5 5 function 2 5 5 PN 4237003C INDEX 0 Quad 20 40 60 Microsecond Delay card jumpers 3 9 4 Quad 20 40 60 Mi
513. to 4 3 1 main system transformer 240 Vac to 4 3 1 inspection Autoclone Sorting Option See Autoclone Sorting Option installation Elite software See software Enterprise laser location illustration of 3 14 3 hardware configuration of 5 PMT 3 11 1 HeCd laser covers illustration of 3 7 12 HeCd laser mounting illustration of 3 7 7 house air 3 8 1 1305 laser mounting illustration of 3 7 9 laser power requirements 3 1 2 of Argon laser illustration 3 7 5 of Autoclone Sorting Option 3 10 1 of beam separator location with Enterprise laser illustration 3 14 3 PN 4237003C of beam translator upgrade 3 12 1 of collection optics 3 15 1 of deflection plate 3 5 3 of dual CRT 3 2 2 of Enterprise laser 3 14 1 of Gated Amp 3 9 1 of house air 3 8 1 of HP 1600C color printer 3 20 1 of HP 1600C color printer software 3 20 2 of instrument 3 2 4 of laser covers water cooled illustration 3 7 13 of optional lasers 3 7 1 of Piezo bimorph 3 16 1 of Piezoelectric transducer See Piezo of PMT 5 3 11 1 of sample collection compartment 3 2 7 of software version 4 0 3 18 1 of software version 4 01 3 19 1 of software general 4 9 1 of sort kit 3 13 3 of sort upgrade 3 13 1 PMT 5 software requirements 3 11 1 requirements for system installation 3 1 1 system power requirements 3 1 2 waste bottle modification software version 4 01 3 19 2 waste bottle modification software version 4 01 illustration of 3 19 3 instr
514. to acquire PMT2 PMT2 Peak and Time of Flight Press On the Cytometer touchscreen a Press Options b Under Align LED select Pulsed 100 0 25 kHz Place the mirror filter holder in the dichroic 45 degree slot below PMTI Remove the 525 nm filter below PMT2 Adjust PMT2 HV and gain to obtain pulses on the Cytometer scope display for PMT2 Peak of about 5 V two divisions amplitude Note the Time of Flight blocks on the Cytometer Options screen TOF should be on and Time base should be 10 0 microseconds Note Check your protocol if your readings are incorrect Verify that the protocol Cytosettings are set to REC then press to restart acquisition Observe the TOF histogram Adjust the Beam Width Normalization control to shift the distribution until the histogram mean channel corresponds to the PMT2 Peak pulse width estimate based on Cytometer scope display width of pulse at 1596 above baseline For example For an observed pulse width of 6 microseconds and a TOF time base of 10 microseconds the histogram mean channel is 6 16 1024 614 6 5 MAINTENANCE PROCEDURES VERIFICATION INSPECTION PROCEDURE 5 1 4 PN 4237003C MAINTENANCE PROCEDURES PREVENTIVE MAINTENANCE PMI PROCEDURE 5 2 PREVENTIVE MAINTENANCE PROCEDURE Purpose The PMI procedure is intended to help prevent failure of the Elite In this procedure you may be required to replace a worn component clean the system or make necessary
515. to pass 26 Adjust the air cooled mirror for optimal FALS signals Note the width of the PMT 2 Peak pulse 27 Open the HeNe and align the air cooled mirror to place the Argon beam on top of the HeNe with FALS or PMT 2 Peak and PMT 4 Peak ATTENTION Do not adjust the beam shaping optics 28 Carefully remove the beam reducer Figure 3 7 11 29 Block the HeNe and air cooled Argon beams 30 Align the UV mirror to obtain optimal signals on PMT 2 Note The beam is out of focus at this point so the signal will be bad 31 Install the beam reducer and adjust it for optimal signals from 2 Peak Adjust the beam reducer focus ring to match the pulse width with the Argon pulse as measured in step 26 3 7 11 INSTALLATION PROCEDURES OPTIONAL LASER INSTALLATION 32 Open the HeNe beam and use only the beam reducer and the PMT 2 Peak and PMT 4 or PMT 5 on 5 PMT systems Peak adjustments to align the UV beam to be on top of the HeNe beam 33 Install remaining covers See Figure 3 7 12 and Figure 3 7 13 34 Install warning labels See Figure 3 7 14 and Figure 3 7 15 Figure 3 7 12 HeCd Laser Covers 1 7003007A 3 7 12 PN 4237003 INSTALLATION PROCEDURES 3 OPTIONAL LASER INSTALLATION Figure 3 7 13 Water Cooled Laser Covers Menem 7003013A 3 7 13 PN 4237003C INSTALLATION PROCEDURES OPTIONAL LASER INSTALLATION V6L0 007 NOLLVIGVY Q3H311VOS HO 1938410 OL NIMS HO 3A
516. tput is compared to the requested 488 power and the error is used to modify the requested UV power The laser cannot be operated at 488 power levels which are so low as to provide no UV output The laser should NEVER be operated in either regulation mode such that its UV output is less than 10mW For dual beam use UV regulation appears to be the best choice for optimum UV power stability Allow 1 5 hours for warmup to stabilize the 488 power Installation 1 Uc P o 10 If the Elite system is not already equipped with the Interlock Relay card and large laser compatible covers install the interlock system optical rail and relocate the air cooled lasers Note If using the Enterprise laser the air cooled Argon will not be used therefore remove its dichroic Verify that the output aperture of the Enterprise laser is at hole 12 3 See Figure 3 14 1 Warm the laser up at 50 mW UV for 30 minutes Install two 5 in targets on the table so their apertures are above row three Adjust the laser so the beam passes through the targets The goal is to have the beam parallel with and 5 in cm above the row Clamp the laser feet to the table using the supplied hardware Remove the targets and install the beam separator with two bolts at the position shown in Figure 3 14 2 Install the 5 in targets at 1 3 and 1 6 so the apertures are at these coordinates Set the two beam translators to be vertical and straight to introduce
517. translator can be mounted as in the attached diagram for use when adjusting the beam to the proper height The resultant beam is parallel to the exit beam of the appropriate laser This allows the user to use either co lateral or separate beams depending on the application Tools Supplies Needed 9 9 9 9 Beam Translator Kit PN 6912985 4 Laser alignment targets with three target holes large PN 6857274 6 small PN 6857236 3 Mounting screws PN 2851920 6 Fluorospheres Laser Mounting Procedure The long dimension of the optical bench is referred to as the columns The shorter dimension of the optical bench is referred to as the rows 1 Mount the UV laser in the farthest rear position possible on the table top Note Allow enough room to adjust the laser XY axis column 16 row 9 parallel with the long dimension of the optical bench Allow enough room in front of the laser for optics including the beam expander reducer a target and the optical rail Mount the Argon laser in the column 8 9 and 10 row 7 Note Allow enough room in front of the laser output for the beam expander a target and the optical rail Mount the HeNe laser next to the fluorescence collection assembly Note Allow enough room for the alignment of the XY axis of the HeNe laser and the fluorescent collection assembly Allow enough room in front of the laser output for the beam expander a target and the optical rail Standard Laser Align
518. ts of the behavior of light are simplified to provide you with a basic understanding with minimum mathematical complexity Light PN 4237003C Definition Light is a form of electromagnetic radiation with wavelengths that range from about 200 to 4 000 nanometers nm Wavelengths from about 400 to 700 nm are visible by human eye Wavelengths shorter than 400 nm are ultraviolet UV Wavelengths longer than 700 nm are infrared IR A unit photon of light is produced when an atom changes energy levels In most light sources many atoms change energy levels randomly In modern optics light consists of a dual nature In some cases light behaves as a wave and in other cases it behaves as a massless energy carrying particle a photon Media Used for Travel Light travels through various media such as air glass and water Each medium has a property known as index of refraction n where n is the ratio of the speed of light in air compared to that in the specific medium As light travels obliquely from one media into another the change in n changes the direction of the light this is known as refraction The principle of refraction makes it possible to trace the path of light rays and to understand the action of lenses Refer to Lenses in this section for more information Figure 2 7 1 shows the refraction of a light ray as it passes from air into glass Notice that the ray is bent toward a line perpendicular to the glass surface Figure
519. ttom of the second to the last attached drop 3 View the side streams periodically while sorting to ensure that they are clean and stable Adjust the drive while watching the droplet in the camera to ensure the connection between the last drop and the second to last drop is not severed Note Optimal side streams are evident when the filament between the two drops is very thin and remains connected 5 Ensure the side streams remain stable Ifthe side streams remain stable go to step 6 Ifthe side streams appear to slightly fan adjust the crystal drive as follows gt Ifthe filament connecting the last attached droplet breaks decrease the crystal drive gt If the filament connecting the last attached droplet fattens increase the crystal drive gt Ifthe side streams remain unstable stop the sort and investigate the problem 6 Press DEBUBBLE and or CLEAR three times e Ifthe droplet returns to the same place the sort should be fine Ifthe droplet does not return to the same place then investigate further before proceeding with the sort otherwise the purity and recovery may be affected 7 Ifyou are performing this procedure in sequence beginning with Acquisition Check return to step 7 under the Sort Check procedure PN 4237003C 3 6 7 INSTALLATION PROCEDURES SYSTEM TESTING 3 6 8 PN 4237003C INSTALLATION PROCEDURES OPTIONAL LASER INSTALLATION PART B OPTIONS AND UPGRADES INSTALLATION 3 7 OPTIONAL
520. uad peak sense and hold QUAD 7 DLY Quad 7 Microsecond Delay card QUAD 20 DLY Quad 20 40 60 Microsecond Delay card R R potentiometer resistor right research RAM random access memory REF reference REG regulator RF radio frequency RG pressure regulator ROM read only memory S Scat scatter Scat Aux Scat Aux Gated Amp SCSI small computer system interface SD standard deviation Sensor I F Sensor Interface SIMM single in line memory module SL solenoid SN serial number SOL solenoid SP short pass stretched pulse SS side scatter SUP supply SVGA super video graphics adapoer SVP system verification procedure SW switch SW R ABBREVIATIONS 3 ABBREVIATIONS sync synchronized T TEMP temperature TOF time of flight TP test point trans translator TRIG trigger U L Underwriters Laboratory NC unified coarse thread NS unified special thread PPBM upper beam U U U U UT digiscope upper trace BNC connector U V ultraviolet V V volt 4 ABBREVIATIONS Vac voltage alternating current vac vacuum VC vacuum chamber Vdc voltage direct current video graphics adaptor VL valve volt voltage VS Versus W W watt w with WM wire marker w o without wt weight X x multiply by xmit transmit PN 4237003C Numerics 0 zero value defined 2 13 1 3 PMT Sub Amp 1 c
521. uisition lower 2 5 6 Gated Amp upper 2 5 6 lower defined 2 5 6 PMT power supply back 2 5 6 PMT power supply defined 2 5 6 upper defined 2 5 6 CC marker See Autoclone Sorting Option 2 2 3 CGI CFG file editing for HP 1600C color printer for DOS 6 0 and higher 3 20 2 lower than DOS 6 0 3 20 4 See also HP 1600C color printer channel 1023 function of 2 13 4 channels See acquisition channels See also channel 1023 circuit Camera Motor Control function of 2 5 8 peak sense and hold function of 2 13 3 2 13 4 circuit cards 20 40 60 Microsecond Delay See 20 40 60 Microsecond Delay card 256K Memory See 256K Memory card 3 PMT Sub Amp SW R See 3 PMT Sub Amp SW R card PMT Sub Amp SW RI See 3 PMT Sub Amp SW Rl card 3 PMT Sub Amp SW R2 See 3 PMT Sub Amp SW R2 card 3 PMT Sub Amp See 3 PMT Sub Amp card 3 PMT Sub SW R See 3 PMT Sub SW R card 7 Microsecond Delay See 7 Microsecond Delay card ADC and PSH Control See ADC and PSH Control card ADC See ADC card all configurations used in 2 10 1 Autoclone Sorting Option Interconnect See 4 INDEX Autoclone Sorting Option Interconnect card Autoclone Sorting Option See Autoclone Sorting Option card Bitmap and Sort Decision See Bitmap and Sort Decision card Camera Interface See Camera Interface card CPU 835 Intel See CPU 835 Intel card CPU IBC 86C See CPU IBC 86C card Cytometer CPU See Cytometer CPU card DAC See DAC card Data Lister Out See Data
522. uisition cycle begins when at least one channel senses a cell s presence The system stretches and holds all pulses for the event The ADC converts stretched pulses to digital values and sequentially places theses values on the data path bus At this time a frame of sequential values on the data path bus represents the event The following can read the digital values as they appear on the bus Prism and Sort Window Test card Bitmap and Sort Decision card and Data Lister Out card The Multibus CPU card writes values to the registers of the cards used in acquisition determining how they will function during acquisition The CPU has nothing to do with the acquisition cycle except to turn the acquisition process on and off The sensors simultaneously measure each cell that passes through the laser The Elite correlates the data that comes from the sensors The value of 0 is a valid piece of information because is means that the particular cell had no fluorescence or no light scatter It is important that the voltage value from a particular parameter remains associated with that particular cell To achieve correlation the Elite uses one channel to detect the presence of cells in the laser beam and then commands all other channels to measure what they see at that time This process is called forced conversion The system must compensate for slight timing differences that occur for different types of parameters The ADC and PSH Control card wo
523. ument description 2 1 1 function 2 1 1 installation procedure 3 1 1 setup 3 1 1 instrument setup 3 2 1 instrument main components illustration of 2 1 1 integration description 2 6 4 integrator circuit function 2 6 4 integrator charge time of 2 6 4 integrator charge time of 2 6 4 integrators 2 6 5 Interface and Scaler R card jumpers 4 10 3 A 2 3 Interface and Scaler card function 2 8 7 Interface and Scaler R card jumpers 4 10 3 A 2 3 Interlock 4 12 1 interlock defeat bar location 4 12 1 IR See infrared defined PN 42370036 INDEX J jumpers for cards in Data Acquisition card cage A 2 3 for cards in Gated Amp card cage 4 10 2 A 2 3 for cards in Multibus Card cage 4 10 1 for cards in Multibus card cage A 2 2 for cards in Non Gated Amp card cage 4 10 2 for line voltages 4 3 1 for Watchdog timing 3 17 1 line voltage A 2 4 non Gated Amp card cage circuit cards in A 2 2 various configurations of 4 10 1 See also circuit cards K KIR3 card beeper Piezo bimorph function of 2 5 5 function 2 5 5 LEDs function of 2 5 5 L Laser Control subsystem description 2 5 2 laser safety ANSI Standard 136 1 Safe Use of Lasers 1 2 2 precautions to follow 1 2 2 warning labels 1 2 3 laser shutter valve VL2 function of 2 9 2 laser shutter location 4 12 5 lasers alignment procedure for Gated Amp instruments 4 1 4 alignment procedure for standard laser configuration 4 1 1 beam sepa
524. up lens The flow cell lens focal length is greater than the distance between it and the intersection point Used this way the lens acts as a magnifier for the pickup lens Because the flow cell lens is close to the light source it intercepts a greater percentage of the total radiated light and relays the light to the pickup lens 27 8 42370036 INSTRUMENT DESCRIPTION 2 SORT SUBSYSTEM 2 8 SORT SUBSYSTEM Function The Sort subsystem Figure 2 8 1 Evaluates each event against previously loaded sort conditions Generates the appropriate signals for droplet formation and droplet deflection Overview Independent Operation Before the sort operation begins the Multibus CPU card sets up the cards for the Sort subsystem During the actual sorting process the Sort subsystem operates without CPU intervention in response to each event Sort Cycle The sort cycle goes through the following processes The cell passes through the laser beam The Acquisition subsystem indicates this time with the DISCSAT signal usually originating in the Quad PSH card it can also originate in the Gated Amp Control card and the Pulse Pileup Det TOF card The acquisition electronics measure selected parameters and place the digitized parameters on the data path The sort electronics perform the following sequence for each cell gt Tests the parameter values for linear gates gt Tests the parameter values for bit maps gt Tests
525. upper beam at flow cell should be 1 32 in lower This means the beam will clip the bottom hole of both targets For Elite Analyzer instruments the front laser upper beam at flow cell should be 1 32 in higher This means the beam will clip the bottom hole of both targets Install small targets on optical plate Remove FALS detector and laser shutter With the lens block and flow cell moved to their highest position of travel aim front laser delayed signals so that back target just clips beam on bottom of hole and front target just clips the beam on the top of the hole Use 488 dichroic mirror adjustments to target Aim the rear laser delayed signals directly through both targets using UV dichroic mirror adjustments Install the beam reducer expander and re target the rear laser beam using the beam reducer expander position adjustments Ensure that beam passes through center of telescope lens Lower beam shaper lens into the beam path and recheck the beams especially for side to side position The beam shape on the back laser target will be elliptical not round Use beam shaper Y and Z axis adjustments to target beams as in steps 4 and 5 Lower the flow cell into the beam path and reposition the flow cell mounting plate as necessary to ensure sample stream is intersected by beams DO NOT adjust beam shaper Z axis to do this Remove targets and reinstall FALS detector and laser shutter Shut off back laser Close aperture
526. ure R1 is 4 99 kohm Ensure C109 is 5 pF Ensure that near U23 U16 U11 07 a diode was added to the card near each of these op amps Gated Amp Control 6705327 Ensure diode location CR5 is not present 3 13 2 See Table 3 13 2 for additional information Table 3 13 2 Additional Information on Cards Card Name Description Minimum Revision Level Peak ADC Replaces ADC card PN 6704139 B Peak Scatter Sensor Located on scatter sensor B Gated Amp Control In all units C Scat Aux Gated Amp Gated Amp units only D PMT Gated Amp Gated Amp units only D Quad PSH 2 used F 3 PMT Sub Amp 1 3 used B Peak Scatter Mux In all units C PN 4237003C PN 42370036 INSTALLATION PROCEDURES SORT PERFORMANCE ESP UPGRADE Sort Kit Installation ale so 11 12 13 14 Do Heading 5 1 VERIFICATION INSPECTION PROCEDURE to ensure instrument is 10096 operational a Record all HV and Gain settings Perform actual sorts with beads Cyto Trol Control cells or cells to determine baseline sorting capability of the unit before beginning upgrade c Make printouts of pre and post sort histograms indicating purity and sort acquisition rates Remove the Sort Delay card and replace it with the Sort Delay R3 card Remove the Sort Oscillator card and replace it with the Sort Oscillator R2 card Install the a Pulse Pileup Det TOF card b Sort bracket c Control panel T
527. urn the instrument off and remove the right front panel to expose the card racks Insert Pulse Pileup Det TOF card in bottom Data Acquisition card cage slot 5 a b Install coax cables as shown in Figure 3 13 1 Replace existing cards with cards in upgrade kit 5 Remove the two upper right screws securing the lower card rack and remove the lower right screw securing the upper card rack Use the screws removed in step d to secure the new control panel Connect the multipin connector to the Sort Oscillator card R2 Connect the cable terminating with a bullet connector to the TOF PPU card J7 Temporarily hold the new front panel up to the instrument and verify that the new cutout in the panel provides access to the controls when the panel is in place Verify instrument operation a Do TOF Verification under Heading 5 1 b Do Heading 4 2 SORT WAVEFORM VERIFICATION AND ADJUSTMENT PROCEDURE c Do Sort Coincidence Verification under Heading 4 2 Install the front panel Install the Piezoelectric Transducer See Heading 3 16 PIEZOELECTRIC TRANSDUCER INSTALLATION for the Piezo assembly included in the sort kit Note If the Piezo is already installed return the part from the Sort kit Do Operational Test Cytometer Electronics Optics or Fluidics under Heading 5 1 Note FALS gain setting must be set higher to achieve the same mean channel obtained before the update You will also note differences in the fluorescence
528. ust translator height in post for laser to penetrate as close to the center of the translator window as possible Refer to the multi hole target and set the Alignment mode for top 1 Move the pin on bottom of target to line up with the desired target hole and lock down with a setscrew Install the middle target in the space between the beam translator and the HeNe dichroic and ensure the target is square with the table top Rotate the translator window until the HeNe passes through the middle of the target hole Remove the HeNe laser target Use the adjustable HeNe laser dichroic on the optical rail look at the lower trace on the Digiscope Screen and adjust the trace between the peak of the HeNe laser caused signal 3 12 5 INSTALLATION PROCEDURES BEAM TRANSLATOR UPGRADE 3 12 6 11 12 13 14 15 16 1T 18 19 20 21 22 23 24 25 and the peak of the UV laser caused signal The adjustment should be e 20 microseconds for a 100 pm quartz tip 30 microseconds for a 76 quartz tip 3 5 microseconds for a 76 pm jet in air tip Note The UV signal is on the far left of the Digiscope Test this by increasing and decreasing the UV laser power and observe the far left peak that follows Close the HeNe laser output shutter Install the adjustable beam translator between the Argon laser and its adjustable dichroic Note Allow enough room between the translator and the optic
529. ustments A 3 2 PN 4237003C QUICK REFERENCE INFORMATION A 1 TOLERANCES AND LIMITS Pressure Compressor Module System pressure 30 psi Compressor pressure 50 psi House Air System Pressure Gauge must not exceed 30 psi System vacuum gauge should read at least 10 in Hg Pneumatics Sheath pressure 12 psi Sample pressure 12 psi Switches DVM set to DC V DVM 4 to 5 V immediately after you turn instrument on 0 V after Compressor module pressure builds up Initial Installation Settings Argon Laser Light output 20 second delays Camera Zoom setting minimum Newer camera assemblies 1 0 Older camera assemblies 0 7 Front and Back Porch Settings for Best Streams Non ESP systems adjust the Sort Oscillator card to make the front and back porch adjustments e Adjust R93 for side streams Keep the front porch level between 60 and 95 of the main pulse e Adjust R92 for center waste stream Keep the back porch level between 5 and 30 of the main pulse PN 4237003 1 1 QUICK REFERENCE INFORMATION TOLERANCES AND LIMITS Test Points Pneumatic Interface Card Table A 1 1 Pneumatic Interface Card Adjustments Adjustment on Card Adjustment Range Test Point R31 10 00 0 01 Vdc TP5 R34 10 00 0 01 Vdc 1 R81 10 00 0 01 TP10 R54 10 00 0 01 Vdc TP3 R51 8 00 0 01 Vdc TP2 Power Supplies Test the Deflection power supply J2 Blac
530. utput R PN 6319584 Sort Transistor card PN 6317927 indicates not used in Analyzer system f ESP indicates new or changed for the ESP system SCHEMATICS SCHEMATICS REQUIRED Gated Amp Card Cage Upper 3 PMT Sub SW R Dual FL Switch R card Gated Amp Backplane Gated Amp Control R 3 ESPt Peak Scatter MUX SW R SCAT CV SW R Sensor Interface RP f ESP indicates new or changed for the ESP system Multibus Card Cage Camera Interface R DT Interface R DIGI Scope Dual CRT Controller Dual Laser Controller Extended Memory Serial C indicates not used in Analyzer system Interconnect Diagrams Interconnect Diagram AC L N area Interconnect Diagram AC RH area Interconnect Diagram DC 400W Switcher Interconnect Diagram DC Distribution Interconnect Diagram DC Sensing area mg mg mg mg cu mg mg mg mg mg mg mg mg mg mg 2122 2 2 24 212 2 2 2 24 2 2 2 2 Z 320686 6320687 6319579 6320758 6320829 6320688 6319759 6319828 6319006 6319024 6319472 6319732 6323668 6317988 6320236 6320233 6320235 6320289 6320238 42370036 PN 42370036 Other 36 VDC Power Supply board A board of Lister AT Analyzer High Voltage DAC B board of Lister AT HV DAC PMT backplane KIR3 Motor Controller R Peak Scatter Sensor card Pinhole LED Driver PMT Connector Board R Pneumatic Interface R2 Pump Motor Det tube sensor Sort HV Deflection Supply Switch amp
531. uts to be FL2 OUT subtracted Programmed Divider Outputs to be subtracted N V J7 7 PROGRAMMABLE PEAK OUT Programmed Divider GAIN AMPLIFIER Outputs to be subtracted gem y INTEGRATOR 4 4 Programmed Divider ra J6 Outputs to be FL3 INTEGRAL subtracted PROGRAMMABLE OUT GAIN AMPLIFIER CONTROL DECODE COMMANDS J8 DIVIDERS CPU uem FL3 LOG OUT TO SET SET INTEGRAL LOGAME SUBTRACTION CARD FUNCTIONS CAIN t SET INTEGRATOR SPEED BANDWIDTH 211 2 7003055 1 42370036 INSTRUMENT DESCRIPTION 2 CARD DESCRIPTION NON GATED AMP NON SWITCH CONFIGURATION EXCEPT ANALYZER Dual FL Amp Card Function The Dual FL Amp card processes each input to provide variable gain peak and integral signals as well as a log output See Figure 2 11 3 Inputs The Dual FL Amp card accepts two FL inputs FL1 and FL2 Figure 2 11 3 Block Diagram Dual FL Amp Card J5 PROGRAMMABLE GAIN J2 FL1 IN AMPLIFIER FL1 PEAK OUT INTEGRATOR PROGRAMMABLE GAIN D AMPLIFIER MD Hi LIN OUT Dit LOG AMP E LOG OUT J7 PROGRAMMABLE GAIN ti J1 FL2 IN AMPLIFIER FL2 PEAK OUT INTEGRATOR Ei PROGRAMMABLE
532. w cell on the following instruments Elite 2362 Elite ESP 2363 Refer to the Special Procedures and Troubleshooting manual for additional information on flow cell adjustments Tools Supplies Needed M DOODO Elite ESP sort sense flow cell 76 p 6859313 1 Elite ESP sort sense flow cell 100 p PN 6859300 0 Sort sense flow cell 76 p PN 6856511 1 Sort sense flow cell 100 u PN 6856101 9 Barbed fittings 5 0 115 i d to 0 180 PVC PN 6232418 0 Barbed fittings 5 6 for instruments with Autoclone Sorting Option 0 062 i d to 0 062 PVC PN 6232417 1 Sheath filter 0 2 m with fittings PN 6858525 2 Flow cell body with fixed insertion rod PN 6858366 7 DVM 50 bleach solution 2 L Fluorospheres IsoFlow sheath fluid PN 8547008 6 Deionized water Microscope gt Fluorescent microscope is recommended gt Light microscope if fluorescent microscope is not available Procedure ATTENTION The instrument must be in proper working order with no sheath sample system or regulator pressure leaks Before changing the flow cell verify there are no metal barbed fittings on the valve bracket assembly See Figure 4 11 1 If there are metal fittings on this assembly or if there is a blackish greenish buildup where the insertion rod enters the flow cell body do Replacing Metal Fittings with PVC Fittings before replacing and testing the new flow cell PN 4237003C 4 11 1 4 SERVIC
533. w cell tip d Beam shaping lens e Sample tubing f Sample drawer Reconnect a The electrical connector to the bimorph crystal drive b The ground connector at the deflection plate holder Do Heading 5 1 VERIFICATION INSPECTION PROCEDURE Repeat step 1 to do the Sensitivity test Ensure the customer is satisfied with the upgrade 3 15 3 INSTALLATION PROCEDURES COLLECTION OPTICS UPGRADE 3 15 4 PN 4237003 3 16 PN 42370036 INSTALLATION PROCEDURES PIEZOELECTRIC TRANSDUCER INSTALLATION PIEZOELECTRIC TRANSDUCER INSTALLATION Purpose Do this procedure to install the Piezoelectric Piezo transducer in instruments manufactured before March 1993 Tools Supplies Needed Stage plate flow cell kit PN 6912795 9 which includes gt Stage plate flow cell PN 6858242 3 Flow cell with intro rod PN 6858366 7 gt Cap nut PN 1021202 2 gt Tubing silicone 0 015 in 0 04 cm PN 3213154 9 Installation 1 Turn off the instrument 2 Remove the flow cell tip and set it aside 3 From the flow cell remove the a Sheath line b Vacuum line c Sample line Unplug the bimorph crystal drive connector Remove the two screws that secure the flow cell mount to the flow cell stage Use the screws you removed in step 5 to now attach the Piezo assembly to the same threaded holes from which the old assembly was removed 7 Install the new flow body a Attach the new flow body to the Piezo assembly b
534. while in current mode Run beads and establish pulses for both FALS and fluorescence Make all usual adjustments to ensure good CVs and sensitivity Turn on beam from back laser Adjust power on lasers so pulses are nearly equal With DNA check beads this means about 10X power for UV than for visible wavelengths Optimize pulses using telescope controls Adjust peak signals form both lasers so they are 40 microseconds apart for 100 pm quartz and 7 microseconds apart for jet in air Cycle through several adjustments of all alignment controls to obtain good pulses from both beams PN 4237003C PN 42370036 14 15 16 17 18 INSTALLATION PROCEDURES GATED AMPLIFIER UPGRADE If the peak signal pulse width is not very similar for signals from both lasers minimize the pulse width for the front laser by adjusting the beam shaper focus and minimize the rear laser peak signal pulse width by adjusting the beam reducer expander You may have to constantly reposition the back beam with the telescope control at the same time Once both beams look good go to the Gated Amp Control screen and set the clock delay and width Set delay to 40 microseconds for 100 quartz and 7 microseconds for jet in air Set source to some signal associated with the front laser beam Delay all signals associated with this beam Turn on gate window display and adjust accordingly so that the entire peak signal pulse width is enclosed by the window

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