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Agilent 1200 Series Micro Collector/Spotter

1. On completion of this procedure Re install the tray s in the tray base Start the instrument Close the front cover Agilent 1200 Series Micro Collector Spotter User Manual 125 5 Maintenance and Simple Repairs Simple Repairs Procedures 126 Agilent 1200 Series Micro Collector Spotter User M anual Agilent 1200 Series Micro Collector Spotter User Manual 6 Parts and Materials Supported Trays for the Micro Collector Spotter 128 List of Recommended Plates 130 MALDI Spotting Accessories 131 Supported M ALDI Targets 133 Transport Unit Assembly 135 Internal Tray Assembly 136 Micro Collector Spotter Accessory Kit 137 ot Agilent Technologies 127 6 Parts and Materials Supported Trays for the Micro Collector Spotter Table 11 Trays for the Micro Collector Spotter Item Description Part Number 1 Full tray for 4 well plates G1364 84521 2 Full tray for 4 M ALDI plate carriers adjustable G1364 84511 3 Std tray for 2 well plates 10 2ml vials G1367 60011 4 Adapter air channel installed underneath plug channel if G1329 43200 the fraction collector is used with the thermostat 5 Plug channel G1364 47200 Only one type of well plate can be used at a time 128 Agilent 1200 Series Micro Collector Spotter User M anual 6 Parts and Materials Trays Figure 22 129 Agilent 1200 Series Micro Coll
2. 7 Push the capillary through the guiding fitting assembly and attach the wellplate adapter Then adjust the position of the capillary tip For highest accuracy that is required for 384 well plates the tip must be close to the groove in the middle of the wellplate adapter 8 If you are using a deep well plate you should push the capillary through the guiding assembly that the tip is close to the bottom of the wellplate adapter frame The tip must stay within the oval frame 9 If the capillary tip is at the correct position remove the wellplate adapter tighten the clamp nut and remount the adapter ia 0 ee clamp nut 0 Agilent 1200 Series Micro Collector Spotter User Manual 119 5 Maintenance and Simple Repairs Exchanging the Capillary Guiding Assembly Frequency When Capillary guiding assembly is bent or damaged Tools required None Parts required Capillary guiding assembly G1364 87303 To avoid personal injury keep fingers away from the needle area during micro WARNING collector spotter operation Regularly inspect the Fraction Collector Capillary and exchange it if it is worn out or CAUTION shows visible signs of damage Before beginning this procedure Move the transport unit of the micro collector spotter in the Home Positi
3. Use the m key for the context sensitive menu It gives you additional tools for the timetable lTimetable Ra PRA DER Bura o oof Pagay Time Module Setting end of module timetable 14 Geapy Line Paste Ling 3Print Timetabl Insert Agilent 1200 Series Micro Collector Spotter User M anual Control Module Screens of the Fraction Collector Settings With the F3 key Detectors in the Settings screen you can configure and Detectors Enable F8 or Disable F7 any detector that is part of a system for peak detection Changes must be acknowledged with F6 Done Settings Detector Use Upslope Downslope Threshold DA Detector OFF ge RISD for Mosse bened Fraction Coiectiori Fraction is all Peak Detectors detect a peak collected when at least one Peak Detector detects a peak Settings With the F4 key Thermostat in the Settings screen you can set the micro Thermostat collector spotter thermostat temperature Changes must be acknowledged with F6 Done Settings Temperature Cl at Power On turn temperature on Agilent 1200 Series M icro Collector Spotter User M anual 3 75 3 Control Module Screens of the Fraction Collector Settings With the F5 key Runtimes in the Settings screen you can change the stop Runtimes time and the post run time for the selected module individually Changes must be acknowledged with F6 Done Stoptime OFF G 00 m
4. Line voltage Line frequency Power consumption apparent power Power consumption active power 100 240 VAC 10 Wide ranging capability 50 or 60 Hz 5 200 VA Maximum 180 W Maximum Ambient operating temperature Ambient non operating temperature Humidity Operating Altitude Non operating altitude Safety standards IEC CSA UL 4 55 C 41 131 F 40 70 C 4 158 F lt 95 at 25 40 C 77 104 F Non condensing Up to 2000 m 6500 ft Up to 4600 m 14950 ft For storing the micro collector spotter Installation Category Il Pollution Degree 2 For indoor use only 12 Agilent 1200 Series Micro Collector Spotter User M anual Installing the Micro Collector Spotter 1 Table 2 Physical Specifications Thermostat G1330B Type Specification Comments Weight 18 5 kg 40 7 Ibs Dimensions 140 x 345 x 435 mm height x width x depth 5 5 x 13 5 x 17 inches Line voltage 100 120 or 220 240 VAC 10 Wide ranging capability Line frequency 50 or 60 Hz 5 Power consumption active power 210 W Maximum Power consumption apparent power 260 VA Maximum Ambient operating temperature 4 40 C 41 131 F Ambient non operating temperature 40 70 C 4 158 F Humidity lt 95 at 25 40 C 77 104 F Non condensing Operating Altitude Up to 2000 m 6500 ft Non operating altitude Up to 4600 m 14950 ft For storing the micro collector spotter Safety standa
5. fraction 82 raction delays 82 raction filling order 83 raction collector capillary 116 raction delay volumes 43 raction list 57 raction preview 51 raction trigger mode 47 raction vessel capacities and trays 140 raction volume 140 uses 10 f f f f f f f f f f G GLP features 141 H half tray combinations 28 home position 94 initialization 37 installing corrugated waste tubing 20 installing the fraction collector 19 interface cables 19 power cable 19 safety 19 installing the thermostatted fraction collector interface cables 23 power cable 23 preparation 23 safety 19 23 instrument status indicator 91 153 interfaces 141 internet 151 L lamp 37 liquid contact control 35 45 M maintenance 93 maintenance functions 92 home position 94 park arm position 94 parts exchange position 94 transport unit self alignment 95 maintenance positions 93 MALDI spotting 40 50 MALDI Spotting tip 40 MALDI targets 45 manual reset 37 maximum capacity 141 maximum flow rate 141 method 46 setup 46 47 minimum fraction volume 140 missing parts 14 needle assemblies 135 needle movement 44 0 Online 56 operating modes 140 P park arm position 94 park transport unit assembly 32 parts and materials 14 127 needle assemblies 135 plates 130 transport unit assembly 135 parts exchange position 94 peak based 47 peakwid
6. micro autosampler micro well plate sampler column compartment optional diode array detector multiple wavelength detector with 500nlI or 80 nl flow cell micro collector spotter ZORBAX SB C18 5 um 150 x 0 5mm 5064 8256 Check out sample 01080 68704 diluted 1 10 in Acetonitrile 15 ul min 10 minutes 30 HPLC grade water 70 HPLC grade acetonitrile Signal A 254 4nm Reference 360 80nm gt 0 1min 2s 200 nl 25 0 C or ambient Peak based 1min Threshold slope 5 mAU s 5 mAU s 5 mAU 3000 mAU Expected result Run a single injection under the conditions as described in Table 6 You should observe a chromatogram with four peaks and four fractions should have been collected in the well plate Agilent 1200 Series Micro Collector Spotter User M anual Configuration and Operation of the Micro Collector Spotter 2 Check out procedure for M ALDI spotting To check out the MALDI spotting functionality a run with blank injection with time based spotting is performed to verify that Agilent 1200 Series micro collector spotter consistently collects onto the MALDI target Prerequisites V The Agilent 1200 Series capillary LC or nanoflow system has been installed and prepared as described in the Agilent 1200 Series capillary LC system manual G1376 90011 or the Agilent 1200 Series nano LC system quick start guide G2226 90011 V The Agilent 1200 Series micro collector spotter has been installed and confi
7. or 4 x 27 Eppendorf tubes 0 5 1 5 2 0 ml MALDI Target Plates 2 xwell plates std tray MTP 10 x 2 ml vials 1 half tray with 384 or 96 well plates standard and conical shape or 2x 27 eppendorf tubes 0 5 1 5 2 0 ml 96 Agilent plate for AP M ALDI 100 Applied Biosystems 2x96 Applied Biosystems 192 Applied Biosystems 400 Perseptive Biosystems Micromass 80 96 spots Bruker Anchor Chips 384 1536 spots 4 3 for Bruker Anchor Chip 1536 Typically 2 ul depending on the fraction collection container Agilent 1200 Series Micro Collector Spotter User M anual Specifications 7 Table 20 Performance Specifications Agilent 1200 Series M icro Collector Spotter G1364D continued Type Specification MALDI spot size 100 5000 nl depending on the M ALDI plate maximum spotting rate 20 spots min 1spot 3s Maximum flow rate 100 ul min delay volumes ul 25 um ID fraction collector capillary 0 25 50 um ID fraction collector capillary 1 100 um ID fraction collector capillary 5 cooling Recommended with additional G1330B maximum capacity 2 micro collector spotter connected via 2 Position 6 Port micro valve G1162A GLP features Early maintenance feedback EM F electronic records of maintenance and errors interfaces Controller area network CAN optional LAN or external contacts interface RS232C APG remote for remote start stop signals to from other modules Int
8. waste port cleaning 109 waste tubing exchange 122 weight 11 well plates 28 45 130 384 conical well plates 97 154 www agilent com In This Book This manual contains user information about the Agilent 1200 Series micro collector spotter The manual describes the following e configuration and operation e control module features e troubleshooting and test functions e maintenance and simple repairs e parts and materials e specifications e safety information Agilent Technologies 2006 Printed in Germany 02 06 E1364 80011 G1364 90011 ss Agilent Technologies
9. Agilent 1200 Series capillary LC system After the check out sample 01080 68704 has been injected four peaks have to be detected and four fractions collected Prerequisites V The Agilent 1200 Series capillary LC system has been installed and prepared as described in the Agilent 1200 Series capillary LC system manual G1376 90011 V The Agilent 1200 Series micro collector spotter has been installed and configured The delay volume has been specified V A well plate is installed as plate 1 in the 4 well plate tray If 384 well plates are used the Arm Length Calibration on page 97 and the Position Accuracy Calibration for 384 Well Plates on page 100 have to be executed before running the check out V All wells are empty and the fill levels which are stored on the module have been reset To verify this remove and insert the 4 well plate tray Then click on Yes in displayed dialog box Agilent 1200 Series Micro Collector Spotter User M anual 63 2 64 Configuration and Operation of the Micro Collector Spotter Table6 Method parameters for micro fraction collection check out Agilent 1200 Series modules Column Sample Flow Stoptime Solvent A Solvent B Wavelength DAD MWD Peakwidth Responsetime Injection volume Column temperature Fraction trigger mode Max peak duration DAD MWD working mode Upslope Downslope Threshold Upper threshold micro degasser capillary pump 20ul flow sensor
10. Collector Spotter User M anual Safety Information A Lithium Batteries Information Danger of explosion if battery is incorrectly replaced Replace only with the same or equivalent type recommended by the equipment manufacturer Lithium batteries may not be disposed off into the domestic waste Transportation of discharged Lithium batteries through carriers regulated by IATA ICAO ADR RID IM DG is not allowed Discharged Lithium batteries shall be disposed off locally according to national waste disposal regulations for batteries Lithiumbatteri Eksplosionsfare ved fejlagtic handtering Udskiftning ma kun ske med batteri af samme fabrikat og type Lever det brugte batteri tilbage til leverandoren Lithiumbatteri Eksplosionsfare Ved udskiftning benyttes kun batteri som anbefalt av apparatfabrikanten Brukt batteri returneres appararleverandoren Bij dit apparaat zijn batterijen geleverd Wanneer deze leeg zijn moet u ze niet weggooien maar inleveren als KCA Agilent 1200 Series Micro Collector Spotter User M anual 147 A Safety Information Radio Interference Never use cables other than the ones supplied by Agilent Technologies to ensure proper functionality and compliance with safety or EMC regulations Test and M easurement If test and measurement equipment is operated with equipment unscreened cables and or used for measurements on open set ups the user has to assure that under operating conditions the radi
11. Figure 16 Location of Status Indicators Power Supply Indicator The power supply indicator is integrated into the main power switch When the indicator is illuminated green the power is ON 90 Agilent 1200 Series Micro Collector Spotter User M anual Troubleshooting and Test Functions 4 Instrument Status Indicator The instrument status indicator indicates the current instrument condition When the status indicator is OFF and power switch light is on the instrument is in a prerun condition and is ready to begin an analysis A green status indicator indicates the instrument is performing an analysis run mode A yellow status indicator indicates a not ready condition The instrument is in a not ready state when it is waiting for a specific condition to be reached or completed for example front door not closed or while a self test procedure is running The next programmed injection will be inhibited until the not ready condition has been cleared An error condition is indicated when the status indicator is red An error condition indicates the instrument has detected an internal problem which affects correct operation of the instrument Usually an error condition requires attention for example leak defective internal components An error condition always interrupts the analysis and prevents the next run after the current run is finished A flashing yellow status indicator indicates that the module is in the resident m
12. MOTE Remote RS232C CAN cable to previous module Ea h CAN DC Out aoe ii wen EMEA CAN cable to next module Figure3 Cable Connections The micro collector spotter is turned ON when the line power switch is pressed and the green indicator lamp is illuminated The detector is turned OFF when the line power switch is protruding and the green light is OFF To disconnect the micro collector spotter from the line unplug the power cord The power will supply still uses some power even switch at the front panel is turned OFF 22 Agilent 1200 Series Micro Collector Spotter User M anual Installing the Micro Collector Spotter 1 Installing a Thermostatted Fraction Collector Preparation Locate bench space Provide power connections Unpack the micro collector spotter and the thermostat Parts required Fraction Collector and thermostat Power cords ChemStation and or Control M odule G1323B 1 Place the thermostat on the bench 2 Remove the front cover and route the condensation drain tube to a waste container Make sure the condensation drain tube runs down into a waste container without any upwards bends or curves Free and unrestr
13. Micro Collector Spotter M ethod Setting up a Micro Collector Spotter M ethod To setup the method parameters of the micro collector spotter select Setup Fraction Collector from the Instrument menu or right click on the fraction collector icon in the graphical user interface This will open the Setup Fraction Collector dialog box as displayed in Figure 6 In the Setup Fraction Collector dialog box general method settings are specified Set up Fraction Collector Instrument 1 E xj Fraction Trigget Mode on Une Timetable Peak bared Max Peak Duration 1 0 min Pesk Detectors Fraction iz collected when a peak is detected by F Use MSO for Mare bared Fraction Collection all selected peak detectors at leatt one solocted peak detector Fenction Proview Max lil volume per location Stoptime bad rong Recovery on the track as configuied 4 wl Posttine on ima Contact Contiol Adjustment 1 0 F Steee temperature signal F Stoe delay sensor signal F Stere UIB signal OK Cancel Help Figure6 Setup Fraction Collector dialog box Fraction Trigger M ode The Fraction Trigger Mode can either be Use Timetable or Peak based can be selected Furthermore for the Peak based mode a time for Maximum Peak Duration can be specified When the Peak based mode is chosen it overrules all Agilent 1200 Series Micro Collector Spotter User M anual 47 2 Configuration and Operation of the Micro Collector Spotter settings in the T
14. System view System view Use the Esc key until you receive Views on the F5 key Choose System from the pull down menu This screen shows the last activities in the system Sample 25 2 Reference l 36 System Use the Esc key to receive Views on the F5 key in the Analysis Screen Choose Configure System from the pull down menu Use the F2 key Configure to select the micro collector spotter Here you define further special set points for the micro collector spotter operation Changes must be acknowledged with F6 Done Tue 16 48 Module Message IAF Collector Front door closed 47 AF Collector Front door open THE 10 25 16 47 00 Id Date Time AF Collector Needle is nearly centeric IME 10 25 16 46 54 AF Collector 1HPLC System 4 Plates NI 10 25 16 46 06 AF Collector B get Preterm reset NI 10 25 16 45 10 80 Agilent 1200 Series Micro Collector Spotter User M anual Control Module Screens of the Fraction Collector 3 Lamp O Time BERCI Idle Use the F1 key Interfaces to access the interface settings if required Changes must be acknowledged with F6 Done ice 0 O08 Ready 4 DO Ouilput LAR Configure trays Use the F2 key Trays to view and configure the automatically detected sample trays Different views will come up depending on the installed trays The standard tray for the micro collector spotter is the 4 wel
15. away from the needle area during micro collector spotter operation CAUTION If solvents with high concentrations of matrix buffer or salt are used the capillary has to be flushed thoroughly with salt free water after analysis Such a procedure prevents the capillary and the waste port from clogging For low flow rates and for MALDI spotting with online matrix delivery we recommend to use the plug to close the waste container instead the waste tubing The plug G1364 26105 is included in the micro collector spotter accessory kit Agilent 1200 Series Micro Collector Spotter User M anual 109 5 Maintenance and Simple Repairs 1The internal tray is located on the right side of the fraction 2 Turn the tray upside down unscrew the tubing and collector It can easily be removed by lifting the clip and remove the washer 3 Now the whole waste vial can be removed as well 4 After unscrewing the cap the waste port can be further disassembled 110 Agilent 1200 Series Micro Collector Spotter User M anual Maintenance and Simple Repairs 5 To clean the wash port and the waste tubing On completion of this procedure For a thorough cleaning of all components they can sonicated for 15 minutes ina ultrasonic bath filled with water Another way to get rid of plugging is to flush the waste capillary from the exit with a water filled syringe Attach the syringe at the end of the waste tubing us
16. fill levels from the previous run D Tray 1 was replaced by a tray of the same type x7 Do you want to reset the fill volume Yes No Figure8 Reset fill volumes The window Figure 8 to reset current fill levels is not displayed if the well plate or the M ALDI target is removed without removing the complete tray Consequently contaminated fractions unwanted overfill events or split fractions might be observed during the following LC run The run time for a system with a fraction collector must be extended by the delay time to ensure the complete collection of all compounds The run time is calculated as run time end of last peak delay time with delay time delay volume flow rate This calculation is only required if the flow rate or the delay volume capillary is changed For further info read Fraction Delay Volumes on page 43 Agilent 1200 Series Micro Collector Spotter User M anual 53 2 54 Configuration and Operation of the Micro Collector Spotter Assignment of Start Location The start location for fraction collection or MALDI spotting can either be assigned in the Sample Info Figure 9 dialog box in the RunControl menu in the Sequence Parameters Figure 10 dialog box or in the Sequence Table in the Sequence menu All fraction start locations are entered in the format micro collector spotter plate number row column e g 1 P1 A 01 with micro collector spotter 1 2 plate nu
17. in a horizontal position If a complete Agilent 1200 Series LC system is to be installed on the bench make sure that the bench is designed to carry the weight of all the modules For a complete system including the thermostatted micro collector spotter it is recommended to position the modules in two stacks see Recommend Stack Configuration for micro collector spotter in the capillary or nanoflow system on page 18 Make sure that in this configuration there is 25 cm 10 inches space on either side of the thermostatted micro collector spotter for the circulation of air Agilent 1200 Series Micro Collector Spotter User M anual 11 1 Installing the Micro Collector Spotter Environment Your micro collector spotter will work within specifications at ambient temperatures and relative humidity as described in Table 1 and Table 2 on page 13 Do not store ship or use your micro collector spotter under conditions where CAUTION na temperature fluctuations may cause condensation within the micro collector spotter Condensation will damage the system electronics If your micro collector spotter was shipped in cold weather leave it in its box and allow it to warm up slowly to room temperature to avoid condensation Table1 Physical Specifications micro collector spotter G1364D Type Specification Comments Weight 13 5 kg 29 8 Ibs Dimensions 200 x 345 x 440 mm height x width x depth 8 x 13 5 x 17 inches
18. on the lamp during the calibration Open the command line in the ChemStation and type microafclampon to switch the lamp ON microafclampoff to switch the lamp OFF WARNING During the arm length calibration the door lock sensor is disabled The fraction collector arm will move while the front door is open In addition the side panel can be removed during this procedure To avoid personal injury keep fingers away from the needle area during arm length calibration All users must be informed about this potential risk To perform an arm length calibration for 384 well plates Step Note 1 Before starting the arm length calibration procedure put two 384 well plates into the front positions 1 and 3 of the well plate tray Select the type of these plates in the fraction collector configuration dialog 2 Switch to the Diagnosis View of the Agilent ChemStation Software Agilent 1200 Series Micro Collector Spotter User M anual 97 4 Troubleshooting and Test Functions To perform an arm length calibration for 384 well plates continued Step Note 10 11 Select Tests from the Diagnosis menu In the Test Selection dialog box select M icro AFC G1364D from the drop down list Select arm length calibration 384 well plates and click on Start Start the arm length calibration and follow the instructions on the screen Click the Plate 1 button in the Arm Length Calibration dialog box Figure 18 on page 9
19. spotter G1364D this volume depends on the installed capillaries The fraction delay volume can be calculated by fraction delay volume volume detector outlet capillary volume fraction collector inlet capillary The volumes of the most common Agilent 1200 Series detector outlet capillaries and the fraction collector inlet capillaries are summarized in Table 1 on page 43 and Table 2 on page 44 To identify the capillary check the label for the part number internal diameter and capillary length For time based operation the detector cell can be bypassed to minimize the delay volume Table1 Detector outlet capillaries description part number length mm 1ID um Volume nl DAD MWD outlet capillary for 500 nl flow cell pre G1315 87338 120 100 945 mounted DAD MWD outlet capillary for 500 nl flow cell G1315 87328 120 50 235 alternative DAD MWD outlet capillary for 500 nl flow cell pre G1315 68708 700 75 3100 mounted old flow cell kit DAD MWD outlet capillary for 80nl flow cell pre 61315 87328 120 50 235 mounted DAD MWD outlet capillary for 80nl flow cell G1315 87318 600 25 300 alternative Agilent 1200 Series Micro Collector Spotter User M anual 43 2 Configuration and Operation of the Micro Collector Spotter Table 2 micro collector spotter inlet capillaries description part number length mm 1ID um Volume nl micro collector spotter capillary 25 um ID 61364 87304 500 25 250 alte
20. the Waste Tubing Frequency When defective or contaminated When waste port is clocked Tools required None Parts required Flap Septum G1364 27107 Waste Tubing G1364 86711 Before beginning this procedure Position the transport unit of the micro collector spotter in the Home Position see Deactivating the door lock sensor on page 92 and remove all installed trays from the tray base Turn OFF the instrument 1 Remove the internal tray from the instrument see Exchanging the Internal Tray on page 124 Don t snap off or squeeze the waste tubing 2 Remove the screw cap from the waste port 3 Exchange the Flap Septum reassemble and tighten the screw cap Agilent 1200 Series Micro Collector Spotter User M anual Maintenance and Simple Repairs 5 4To exchange the waste tubing turn the internal tray upside On completion of this procedure down and unscrew the waste tube fitting from the waste port Assemble the new waste tubing Install the internal tray in the fraction collector as shown in step 4 on page 125 IMPORTANT Make sure that the waste tubing is guided correctly through the waste tubing channel to the waste tubing outlet as shown in step 4 and step 5 on page 125 Agilent 1200 Series Micro Collector Spotter User Manual 123 5 Maintenance and Simple Repairs Exchanging the Internal Tray Fr
21. the micro collector spotter is OFF and the power cables are disconnected 14 Connect the cable between the micro collector spotter and the thermostat see Figure 6 on page 26 WARNING Do not disconnect or reconnect the micro collector spotter to thermostat cable when the power cords are connected to either of the two modules This will damage the electronics of the modules 15 Connect the power cables to the power connectors 16 Connect the CAN cable to other Agilent 1200 Series LC modules 17 If an Agilent ChemStation is the controller it must be connected to the LAN interface 18 Connect the APG remote cable optional for non Agilent 1200 Series instruments 19 Turn ON power by pushing the button at the lower left hand side of the micro collector spotter 20 The exhaust fan will turn ON and remove potential solvent vapor from the inside of the instrument After 2 minutes close the front door Then the Agilent 1200 Series Micro Collector Spotter User M anual 25 1 Installing the Micro Collector Spotter micro collector spotter will start the hardware initialization process At the end of this process the status LED should be green 21 The micro collector spotter is turned ON when the line power switch is pressed and the green indicator lamp is illuminated The detector is turned OFF when the line power switch is protruding and the green light is OFF WARNING To disconnect the micro collector spotter from the lin
22. the needle in such a wap thet ft touches all cone positions of the target acc ately in thea Contes xy position alignment buttons Corrective Values relative to the previous alignment Figure 19 Position Accuracy Calibration Interface for well plates Agilent 1200 Series Micro Collector Spotter User Manual 101 4 Troubleshooting and Test Functions Position Accuracy Calibration for M ALDI Targets The Position Accuracy Calibration for MALDI targets has to be performed if e a different type of MALDI Target is used e the positioning is inaccurate e after mayor repairs e g exchange of transport assembly capillary carrier or mainboard e after a firmware update We recommend to switch on the lamp during the calibration Open the command line in the ChemStation and type microafclampon to switch the lamp ON microafclampoff to switch the lamp OFF During the position accuracy calibration the door lock sensor is disabled The fraction collector arm will move while the front door is open To avoid personal injury keep fingers away from the needle area during position accuracy calibration All users must be informed about this potential risk The position accuracy depends on the temperature We recommend to perform the position accuracy alignment at the same temperature as the operation of the instrument If a fraction collector thermostat is present it should be switched on 30 minutes before the calibration to ensure cons
23. well plate can be used on the well plate tray The checkbox Treat as 4 x 96 is only available for 384 well plates and allows splitting the 384 well plate virtually into 4 separate 96 well plates This only changes the collection order but the numbering of the wells remains as indicated on the plate The filling order of each quarter is as specified in the Collection Order section When the 384 well plate is split into four equal quarters the order of the four plates is the same as displayed in the Installed Plates section Each tray position that contains a well plate has to be checked Agilent 1200 Series Micro Collector Spotter User M anual 45 2 46 Configuration and Operation of the Micro Collector Spotter In the Reserved Locations you have the possibility to specify locations that will not be used for Fraction Collection see Table 3 Table 3 Syntax for the definition of Reserved Locations Location Syntax Description Rows A Row A can t be used A B Rows A and B can t be used A D A B Cand D can t be used A D F Rows A B C D and F can t be used Columns 1 Column 1 can t be used 1 2 Columns 1 and 2 can t be used 1 4 Columns 1 2 3 and 4 can t be used 1 4 12 Columns 1 2 3 4 and 12 can t be used Single locations G12 H12 Locations G12 and H12 can t be used Agilent 1200 Series Micro Collector Spotter User M anual Configuration and Operation of the Micro Collector Spotter 2 Setting up a
24. with well plates and Eppendorf tubes e G1364D Thermostatted option of the micro collector spotter This option can be setup by additionally ordering and installing a G1330B fraction collector thermostat 14 Agilent 1200 Series M icro Collector Spotter User M anual Installing the Micro Collector Spotter 1 Table3 Delivery Checklist for the G1364D micro collector spotter Description Quantity micro collector spotter unit 1 Power cord local M atching the sockets in your country or region 1 Accessory kit see below 1 Table 4 Delivery Checklist for the G1330B Thermostat M odule optional Description Quantity Thermostat M odule 1 Power cord local Matching the sockets in your country or region 1 Accessory kit thermostat module see below 1 Accessory Kits Each shipment contents an Accessory kit with the necessary tools to install the system and to have an operating system e The Accessory kit G1364 68715 shown in Table 5 is shipped with the micro collector spotter G1364D Table5 Micro collector spotter Accessory Kit Contents G1364 68715 Description Quantity Part Number Hex key 2 0 mm 1 8710 2476 MALDI Spotting Adapter 1 61364 83205 Flap Septum 1 G1364 27107 Waste Tubing 1 4 mm ID 1 G1364 86711 Waste tubing 1 2 m 1 5062 2463 CAN cable 1m 1 5181 1519 Agilent 1200 Series Micro Collector Spotter User M anual 15 1 16 Installing the Micro Collector Spotter Table5 Micro collector
25. 00 Series Micro Collector Spotter User M anual 137 6 Parts and Materials Micro Collector Spotter Accessory Kit 138 Agilent 1200 Series Micro Collector Spotter User M anual Agilent 1200 Series Micro Collector Spotter User Manual 7 Specifications Performance Specifications for the Micro Collector Spotter 140 ee Agilent Technologies 139 7 Specifications Performance Specifications for the Micro Collector Spotter 140 Table 20 Performance Specifications Agilent 1200 Series M icro Collector Spotter G1364D Type Specification trigger modes operating modes fraction vessel capacities and trays MALDI Spotting plates pre configured M ALDI Plate Capacity Minimum fraction volume Time slices Peak threshold up downslope Timetable combination of time intervals and peak and Agilent 1200 UV Vis detectors DAD G1315A B MWD G1365 A B are fully supported Other detectors with appropriate delay volumes can be connected through UIB interface Above location Into location Liquid Contact Control The tip of the fraction collector capillary will initially move down to the bottom of the well Then it will slowly move upwards while the fraction is collected The contact control mode enables the micro collector spotter to collect fractions down to 2 ul in well plates or M ALDI spots down to 100 nl e 4 well plates full tray MTP with 384 or 96 well plates standard and conical shape
26. 2CHCl3 Og BCOCl2 2HC1 This reaction in which stainless steel probably acts as a catalyst occurs quickly with dried chloroform if the drying process removes the stabilizing alcohol Chromatographic grade ethers which can contain peroxides for example THF dioxane di isopropylether such ethers should be filtered through dry aluminium oxide which adsorbs the peroxides Solutions of organic acids acetic acid formic acid and so on in organic solvents For example a 1 solution of acetic acid in methanol will attack steel Agilent 1200 Series Micro Collector Spotter User M anual Safety Information A e Solutions containing strong complexing agents for example EDTA ethylene diamine tetra acetic acid e Mixtures of carbon tetrachloride with 2 propanol or THF Agilent Technologies on Internet For the latest information on products and services visit our worldwide web site on the Internet at http www agilent com Select Products Chemical Analysis It will provide also the latest firmware of the Agilent 1200 Series modules for download Agilent 1200 Series Micro Collector Spotter User M anual 151 A Safety Information Agilent Technologies on Internet 152 Agilent 1200 Series M icro Collector Spotter User M anual A accessory kit 137 accessory kit contents 16 Agilent on internet 151 air circulation 11 arm length calibration 97 battery safety information 147 bench space 11
27. 37 2 Configuration and Operation of the Micro Collector Spotter Configuration of the Micro Collector Spotter Choosing and Installing the Micro Collector Spotter Capillary The micro collector spotter is delivered with three capillaries each having a different inner diameter 25 50 and 100 um The 50 um capillary is installed in the micro collector spotter whereas the 25 and the 100 um capillaries are in the accessory kit see Micro Collector Spotter Accessory Kit on page 137 To optimize the micro collector spotter for different flow ranges the correct capillary has to be selected and installed For micro fraction collection we recommend flow rates below 4 uL min the yellow 25 um i d capillary G1364 87304 delay volume app 0 25 uL flow rates between 4 and 30 uL min the green 50 um i d capillary G1364 87305 delay volume app 1 uL flow rates between 30 and 100 uL min the black 100 um i d capillary G1364 87306 delay volume app 4 uL For MALDI spotting we recommend for overall flow rates LC flow matrix flow lt 8 ul min the green 50 um i d capillary G1364 87305 delay volume app 1 uL for overall flow rates LC flow matrix flow gt 8 ul min the black 100 um ID capillary G1364 87306 delay volume app 4 uL The exchange procedure is described in Replacing Micro Collector Spotter Capillary Assembly on page 117 An Exchange of the fraction collector capillary might also be necessary if
28. 9 Use the left and right button in the dialog box Figure 18 to adjust the capillary tip to the center of the well Click the Plate 3 button in the Arm Length Calibration dialog box Figure 18 on page 99 in the Arm Length Calibration dialog box Figure 18 on page 99 Use the left and right button to adjust the capillary tip to the center of the well After adjusting the capillary this way the determined values are stored by pressing the OK button Click on Explain to get detailed instructions and information The capillary carrier will move to well position A7 on plate 1 The capillary carrier will move to well position A18 on plate 3 It might take some time to store the positions on the micro collector spotter 98 Agilent 1200 Series Micro Collector Spotter User M anual Troubleshooting and Test Functions 4 Arm Length Calibration rm Length Calibration Figure 18 Arm Length Calibration Interface Agilent 1200 Series Micro Collector Spotter User M anual 99 4 Troubleshooting and Test Functions Position Accuracy Calibration for 384 Well Plates We recommend to perform the Position Accuracy Calibration for 384 well plates e if the positioning is inaccurate e after mayor repairs e g exchange of transport assembly capillary carrier or mainboard e after a firmware update e if a different type of 384 well plate is used We recommend switching on the lamp during the calibration Open th
29. Agilent 1200 Series Micro Collector Spotter User Manual ope Agilent Technologies Notices Agilent Technologies Inc 2006 No part of this manual may be reproduced in any form or by any means including elec tronic storage and retrieval or translation into a foreign language without prior agree ment and written consent from Agilent Technologies Inc as governed by United States and international copyright laws Microsoft is a U S registered trademark of Microsoft Corporation Manual Part Number G1364 90011 Edition 02 06 Printed in Germany Agilent Technologies Hewlett Packard Strasse 8 76337 Waldbronn Warranty The material contained in this docu ment is provided as is and is sub ject to being changed without notice in future editions Further to the max imum extent permitted by applicable law Agilent disclaims all warranties either express or implied with regard to this manual and any information contained herein including but not limited to the implied warranties of merchantability and fitness for a par ticular purpose Agilent shall not be liable for errors or for incidental or consequential damages in connec tion with the furnishing use or per formance of this document or of any information contained herein Should Agilent and the user have a separate written agreement with warranty terms covering the material in this document that conflict with these terms th
30. C cable connections 22 capacities 140 capacity 141 capillary 38 cleaning 39 62 109 M ALDI spotting 38 micro fraction collection 38 replacing 116 capillary guiding assembly exchange 120 capillary tip 39 position 39 check out procedure fraction collection 63 M ALDI spotting 65 ChemStation configuration 42 cleaning the fraction collector 108 collection order 44 condensation 12 configuration 42 collection order 44 fraction delay volumes 43 needle movement 44 trays 43 well plates 45 configuring well plate types 29 Contact Control 45 contact control 45 control module analysis view 69 configure screens 81 detectors 75 EMF 85 error log 87 fraction 82 fraction delays 82 fraction filling order 83 fractions 79 introduction to AFC views and screens 67 maintenance log 87 method screens 76 recovery location 83 reserved locations 83 run times screen 76 sequence 78 settings screens 70 system view 80 tests 84 thermostat screen 75 timetable screens 73 trays 81 cooling 141 corrugated waste tubing 20 D damaged packaging 14 data analysis 57 delay volumes 141 delivery checklist 14 door lock sensor 92 deactivate 92 electrical connections thermostat 26 EMF 85 environment 10 12 Agilent 1200 Series Micro Collector Spotter User M anual Index F ill levels reset 53 ill volume maximum 50 flap septum exchange 122 flow rate 141
31. Calibration Plate for Bruker Targets 5023 0208 Calibration Plate for Applied Biosystems Targets 10x10 20x20 5023 0213 Calibration Plate for Applied Biosystems Targets 192 96x2 5023 0209 Calibration Plate for Micromass Targets 5023 0215 Table 15 Online M atrix Kit G1364 68706 Item Part Number External Contact BCD Board G1351 68701 Cable BCD Board to Syringe Pump 5181 1536 Micro Connector T Type 5042 8519 Agilent 1200 Series Micro Collector Spotter User Manual 131 6 Parts and Materials Table 15 Online M atrix Kit G1364 68706 continued Item Part Number Peek coated fused silica capillary 550mm length ID 125um G1375 87318 Adapter 10 32 to 1 4 28 5042 8517 Syringe 1ml 5181 1541 Union to Luer Fitting 5042 8518 Needle LL 22 51 3 2 pk 5183 4614 132 Agilent 1200 Series Micro Collector Spotter User M anual Parts and Materials 6 Supported M ALDI Targets Table 16 Supported Targets for M ALDI Spotting Note M ALDI target plates for Applied Biosystems Bruker and Micromass are not supplied by Agilent Technologies Dimensions for other targets can be configured in the ChemStation Software Agilent Technologies 96 Agilent for AP M ALDI G1972 60025 Applied Biosystems 80 Applied Biosystems 96 Applied Biosystems 100 Applied Biosystems 96x2a Applied Biosystems 96x2b Applied Biosystems 192 Applied Biosystems 400 Perseptive Biosystems 144 Applied Biosystems 192 Opti TOF tar
32. Simple Repairs Procedures on page 116 WARNING When opening capillary or tube fittings solvents may leak out Please observe appropriate safety procedures for example goggles safety gloves and protective clothing as described in the material handling and safety data sheet supplied by the solvent vendor especially when toxic or hazardous solvents are used WARNING Regularly inspect the capillary and exchange it if they is worn out or shows visible signs of damage WARNING To prevent personal injury the power cable must be removed from the instrument before removing the micro collector spotter cover Do not connect the power cable to the micro collector spotter while the cover is removed Cleaning the Micro Collector Spotter The micro collector spotter covers should be kept clean Cleaning should be done with a soft cloth slightly dampened with water or a solution of water and a mild detergent Do not use an excessively damp cloth from which liquid could drip into the micro collector spotter WARNING Do not let liquid drip into the micro collector spotter It could cause a shock hazard or damage to the micro collector spotter 108 Agilent 1200 Series Micro Collector Spotter User M anual Maintenance and Simple Repairs 5 Cleaning the waste port Frequency The frequency depends strongly on the application Tools required None Parts Required None WARNING To avoid personal injury keep fingers
33. The alignment tools for arm length calibration and position accuracy alignment are not available e Only one fraction start location per sequence can be specified e No tick marks are displayed in the online plot The flexibility of managing data and reporting is very limited Nevertheless the Agilent 1200 Series control module can be helpful during the operation of the micro collector spotter For example the user will be able to trigger the fraction collector manually or specify the flow rate of a non Agilent 1200 Series pump a Agilent Technologies 67 3 Control Module Screens of the Fraction Collector Major keys on the Agilent 1200 Series Control Module ESC Return to previous screen abort any change of parameters and toggle between the last two top layer views m Open context sensitive menus i Information Help Enter Store changed parameters or execute a choice Done If available Activate settings of current screen On Off Switch on individual Instrument s or complete System Start Start a location range or sequence Plot View online signals Views Change between analysis samples status system views The screens shown on the next pages are based on Control M odule firmware revision B 04 0x G1323B HPLC Module firmware revision A 05 06 and higher In case the control module s display seems to be frozen hang up due to a communication problem on the CAN bus unplug the control module from the HPLC module an
34. ack mode is checked the solvent that is delivered between the fractions spots is collected on the same plate for recovery The capillary carrier will move to the next well following collection order e g row by row Agilent 1200 Series Micro Collector Spotter User M anual 49 2 50 Configuration and Operation of the Micro Collector Spotter In the Data Analysis view as well as in the Report a fraction list indicates the wells that contain fractions and the wells that contain recovery With this information the user is able to recover compounds that weren t collected Contact Control Adjustment This parameter is only required for MALDI Spotting If you are adding your matrix online as described in Online Matrix Delivery on page 59 the flow rate at the capillary tip of the micro collector spotter is higher than the flow rate that is measured at the flow sensor of your Agilent 1200 Series capillary or nanoflow pump Consequently the needle movement in the Contact Control mode has to be adjusted The factor can be calculated by Contact Control Adjustment pump flow matrix flow pump flow Some examples are presented in the Table 4 below Table4 Examples for Contact Control Adjustment PumpFlow nl Matrix Flow nl Contact Control Adjustment 500 0 1 0 default 500 500 2 0 1000 500 15 500 1000 3 0 The Contact Control adjustment can also be used to optimize the MALDI spotting process and if the flow is spli
35. act with the liquid surface of the collected fraction This movement types enables the Agilent 1200 Series M icro Collector Spotter User M anual 83 3 84 Control M odule Screens of the Fraction Collector Configure Thermostat System tests Agilent 1200 Series micro collector spotter to collect low volumes without carry over or air bubbles at the bottom of the well Changes must be acknowledged with Done F6 into location above location Options E i contact control mm F4 Therm in the main Configuration screen allows you to configure the temperature and turn on conditions of the thermostatted micro collector spotter Changes must be acknowledged with F6 Done Settings Temperature HA C 26 8 C Cat Power On turn temperature on Press F3 from the System View and choose the micro collector spotter to access the micro collector spotter Tests screen The Tests screen gives you access to multiple tests and features such as the automatic transport unit Alignment F1 tool maintenance procedures to Change F2 parts on the micro collector spotter Parts Info F5 provides information about the type and revision of the installed transport unit assembly The pull down menu in the center of the screen gives you access to various step commands Agilent 1200 Series Micro Collector Spotter User M anual Control Module Screens of the Fraction Collector 3 ice 0 008 Ready Func
36. age 93 Turn OFF the instrument Unscrew the micro collector spotter capillary from the flow cell of the detector Agilent 1200 Series Micro Collector Spotter User Manual 117 Maintenance and Simple Repairs 2 Remove the clamp nut and remove the capillary from the clip at the bottom of the carrier assembly Do not adjust the capillary while the clamp nutis tightly mounted on the capillary guide fe ea N clip lt clamp nut 3 Pull the capillary out off the capillary guiding pipe and out off the guiding hole in z arm of the transport assembly Do not tighten the clamp nut without a capillary installed a hole y guiding pipe 4 Press the metal sheets of fixation spring together and push the capillary assembly out of the holder at the z arm of the transport assembly 5 Unplug the capillary assembly from the bolt carrier located on the top center of the front cover by moving the snapper as indicated by the arrows Slide the Capillary out to the bottom 118 Agilent 1200 Series M icro Collector Spotter User M anual Maintenance and Simple Repairs 5 6 Push the new capillary through the bolt carrier as shown in step 5 of this procedure Then attach the fixation spring at the z arm of the transport assembly and push the capillary through the guiding hole
37. al Control Module Screens of the Fraction Collector 3 fraction collector will move to the next position With this functionality no eluent is flushed to waste and the sample can be recovered from the well plate if it hasn t been correctly collected Recovery on Track is recommended for valuable samples and if the chromatographic behavior of the sample is unknown e g during method development Changes must be acknowledged with F6 Done With the F2 key Timetable in the Settings screen you can list the timetable Settings for the micro collector spotter Press F7 key Insert to edit the selected entry Timetable or F6 key Delete to remove the selected highlighted entry Timetable IE Dea ces OOS Ready Time Module Setting end of module timetable 5_ lt Insert Edit each entry line according to your needs by entering a time for a Setting and by selecting a Setting with the help of the pull down menu Use F7 Enter to insert the current settings Changes must be acknowledged with F6 Done Agilent 1200 Series Micro Collector Spotter User M anual 73 3 74 Control M odule Screens of the Fraction Collector etable BE CRETA ours lt miu 0 cojte Ready Setting Mode Time A min t fields change with selling sefected I Timetable HG loka Matos 0 ooe Ready Time Module Setting 2 00 AF Collector Trigger Mode 0 50 min each fraction 10 00 AF Collector Trigger Mode Off
38. anging the Internal Tray on page 124 Exchanging the flap septum and When defective or contaminated See Exchanging the Flap Septum and the waste tubing the Waste Tubing on page 122 R 116 Agilent 1200 Series Micro Collector Spotter User M anual Maintenance and Simple Repairs 5 Replacing Micro Collector Spotter Capillary Assembly Frequency When contaminated blocked worn out or visibly damaged When using a new capillary with different diameter Tools required None Parts Required Capillary Assembly 25 um ID G1364 87304 Capillary Assembly 50 um ID G1364 87305 or Capillary Assembly 100 um ID G1364 87306 WARNING To avoid personal injury keep fingers away from the needle area during micro collector spotter operation WARNING Thoroughly follow the described installation procedures to maximize the lifetime of the fraction collector capillary and to avoid potential spills or fraction losses Regularly inspect the capillary and exchange it if itis worn out or shows visible signs of damage Before beginning this procedure 1 Remove the wellplate adapter from the Capillary Carrier Assembly Position the transport unit of the micro collector spotter in the Home Position see Maintenance Functions on page 93 Remove all installed trays from the tray base Position the transport unit of the micro collector spotter in the Change Parts Position see Maintenance Functions on p
39. arameters given in the Peak Detector table Additionally a Maximum Peak Duration in minutes has to be specified This parameter can be used to stop the fraction collection in cases where the baseline drifts and the signal does not drop below the specified threshold value The limits are from 0 1 min to 9999 0 min This parameter is mandatory if you use Peak Controlled fraction collection but is disabled for Time Based fraction collection Timetable When the Time based mode is chosen two different options are available e A specific number of fractions or time slices can be specified The of Fractions refer to the number of fractions that are collected in the corresponding time frame in the table This parameter is applicable only for time based fraction collection The maximum spotting rate is 20 spots min e Alternatively time slices can be set up that define the collection time for each fraction This parameter is only available in the time based fraction collection mode Based on the maximum spotting rate of 20 spots per minute the minimum time slice for one spot is 3s e Contact Adj is a parameter which is only required for MALDI Spotting For details please read Contact Control Adjustment on page 50 To edit the Timetable the functions Insert Append Cut Copy and Paste are available To access the additional sections in the Setup Fraction Collector dialog box click the More button Recovery If the Recovery on the Tr
40. asily be graphically specified A chromatogram e g a pilot run can be loaded by pushing the Load Signal button Parameters can now be changed either Agilent 1200 Series Micro Collector Spotter User M anual 51 2 Configuration and Operation of the Micro Collector Spotter manually in the Detector Table and Timetable or graphically in the Fraction Preview screen By pushing the desired buttons on the right hand side of the Fraction Preview screen the chromatogram can be zoomed the values for up and down slope can be specified and the upper and lower threshold level can be set up The graphically specified values are automatically transferred to the Peak Detector Table 52 Agilent 1200 Series Micro Collector Spotter User M anual Configuration and Operation of the Micro Collector Spotter 2 Starting your run with fraction collection or M ALDI spotting Resetting the current fill levels To protect the collected fractions spots for contaminations and to avoid overfilling of single positions the Agilent 1200 Series micro collector spotter stores current fill levels on the module and blocks used positions If a tray with new well plate or MALDI target is inserted in the micro collector spotter all fill levels can be reset by removing and re installing the 4 well plate tray not just the well plate or MALDI plate carrier After re installation of the tray a pop up window as displayed in Figure 8 will be displayed Click on Yes to reset all
41. ble 1 With the capillary pump Time 0 5 minutes Trigger mode Time based of fractions 20 Time 2 5 minutes Trigger mode Off 2 With the nanoflow pump Time 0 5 minutes Trigger mode Time based of fractions 10 Time 5 5 minutes Trigger mode Off Expected result Run a single injection under the conditions as described in Table 7 You should observe 20 spots with the capillary LC system or 10 spots with a nanoflow LC system The spots should be consistently aligned on the MALDI target Due to evaporation the spots collected earlier during the run will be smaller than the later spots Agilent 1200 Series Micro Collector Spotter User M anual Agilent 1200 Series Micro Collector Spotter User Manual 3 Control M odule Screens of the Fraction e Collector Major keys on the Agilent 1200 Series Control Module 68 Screens available from the Analysis view 69 Screens available from the System view 80 This chapter is intended to introduce the user to the functionality for operation of the Agilent 1200 Series micro collector spotters G1364D with the Agilent 1200 Series control module This chapter will provide an overview of the control module features Please use the User Manual of the Agilent 1200 Series control module or the i key online help for further details To use the full functionality of the Agilent 1200 Series micro collector spotter the Agilent ChemStation is required With the Agilent 1200 Series control module
42. capillary or nanoflow LC systems With these systems the collection of very low volume fractions requires low delay volumes high position accuracy of the capillary tip liquid contact control for reproducible collection of small fractions without carryover The principles of operation for the Agilent 1200 Series micro collector spotter will be illustrated in this chapter along with detailed descriptions on how to set up the method and change the configuration This will enable the user to run the instrument with optimal performance The Agilent 1200 Series micro collector spotter designed for low flow rates from below 100 ul min down to 100n1 min In order to keep dispersion a single low volume PEEK coated fused silica capillary is used to direct the flow to the vessel or MALDI target A permanent waste position is built in to allow automated and continuous collection of the eluent The proper alignment of the capillary tip is crucial for precise collection of the fractions especially if 384 conical well plates or MALDI targets are used A sensor pad for automated control of the capillary tip position was therefore integrated into the micro collector spotter A diagram of the fraction collector which indicates the most important components is shown in Figure 1 on page 35 34 Agilent 1200 Series Micro Collector Spotter User M anual Configuration and Operation of the Micro Collector Spotter 2 Waste port Sensor pad In orde
43. ction Delay Volumes the Collection Order the Needle Movement and the well plate MALDI target type can be specified Traction Collector Configuration Instrument 1 x Traps Taya Tube Volume ml D 00 Tube Height mm D Fraction Delay Volumes r Collection Order z Shortest path Detector Volume nl G Row by row n r Column by Ill r Shortest path row Collection Mode Needle Movement Discrete lrachone inta location Depth AL am C above location Distance Z0 am Continuous flow contact control Distance 0 2 mm Well Plates e Pan Treatas fleserved Locations Pie Toe inert a Installed Plates 1 Row s F Plate 1 hrom felt F Plate 3 liront night Coluan s F Piate 2 back left I Plate 4 back right eee ocation s Figure5 Micro Collector Spotter Configuration dialog 42 Agilent 1200 Series Micro Collector Spotter User M anual Configuration and Operation of the Micro Collector Spotter 2 Trays In the online ChemStation the configuration of the Trays is recognized automatically In the offline ChemStation the tray configuration can be chosen in a dropdown menu Fraction Delay Volumes The fraction delay volumes specify the volume between the detector cell and fraction collector capillary tip This volume is required to calculate the time delay between the detection of the peak in the detector and the start of the collection in the fraction collector For the micro collector
44. d the capillary has to be flushed thoroughly with salt free water after analysis Such a procedure prevents the capillary and the waste port from clogging The capillary length has to be adjusted between the following limits see Figure 3 For 384 conical well plates PCR plates where high position accuracy is required the capillary tip should be approximately set to the groove at the center of the well plate adapter When 96 well plates or deep 384 well plates are used the capillary tip must be set to the lower edge of the well plate adapter opening in order to reach the bottom of the wells Agilent 1200 Series Micro Collector Spotter User M anual 39 2 40 Configuration and Operation of the Micro Collector Spotter high position accuracy maximum capillary length e g 384 well plates e g deep well plates Figure3 Positions of the capillary tip Capillary Alignment for M ALDI spotting If the micro collector spotter is used for MALDI spotting the well plate adapter has to be replaced with the spotting adapter G1364 83205 which is supplied in the accessory kit see Table 19 on page 137 For MALDI Spotting applications we recommend using the MALDI spotting tip This tip prevents that the droplet from flowing upwards on the outer capillary wall 1 Remove all trays and move the fracti
45. d alignment plate fits into the slit between the pusher and the adapter t shaped alignment plate 114 Agilent 1200 Series Micro Collector Spotter User M anual Maintenance and Simple Repairs 5 Connect the lamp assembly to the extension cable Push the cable connector in between the yellow plastic clip as described in step 3 of this procedure Plug the lamp power cable into the connector at the front of the z arm Install the fraction collector spotter capillary as described in Replacing Micro Collector Spotter Capillary Assembly on page 117 To activate the lamp switch open the command line in the ChemStation and type microafclampon to switch the lamp ON microafclampoff to switch the lamp OFF Agilent 1200 Series Micro Collector Spotter User Manual 115 5 Maintenance and Simple Repairs Simple Repairs Procedures The procedures described in this section can be done with the micro collector spotter in place in the stack Table 10 Simple Repair Procedures Procedure Typical Frequency Notes Replacing the fraction collector When worn out blocked or damaged See Replacing Micro capillary When using a capillary with different diameter Collector Spotter Capillary Assembly on page 117 Exchanging the capillary guiding When bent or damaged See Exchanging the Capillary Guiding assembly Assembly on page 120 Exchanging the internal tray When flow delay sensor defective See Exch
46. d reconnect 68 Agilent 1200 Series Micro Collector Spotter User M anual Control Module Screens of the Fraction Collector 3 Screens available from the Analysis view Analysis view This is the start up screen if the Agilent 1200 Series micro collector spotter or thermostatted micro collector spotter is the only configured Agilent 1200 Series LC module m Temp 4 c Settings Timetable Method Sequence On Off F7 allows you to turn on the Thermostat Module The m key allows access to the context sensitive menus Date amp Time allows you to change time settings Print Screen sends the screen to a configured printer location About tells you the current firmware revision and the serial of your control module Setup view leads you to analysis view configuration for additional Agilent 1200 Series LC modules Restart re boots the control module If multiple instruments are configured in the Analysis screen the context menu contents will vary depending on the cursor position when the m key is pressed depending on highlighted module Agilent 1200 Series M icro Collector Spotter User M anual 69 3 70 Control M odule Screens of the Fraction Collector Settings Mon 10 56 2Print Screen 3 About Settings Timetable f Sequence In the Setup view modules can be added or removed to the analysis view Select a module you want to move with the help of the selection keys Use function ke
47. e command line in the ChemStation and type microafclampon to switch the lamp ON microafclampoff to switch the lamp OFF During the position accuracy calibration the door lock sensor is disabled The fraction collector arm will move while the front door is open In addition the side panel can be removed during this procedure To avoid personal injury keep fingers away from the needle area during position accuracy calibration All users must be informed about this potential risk 100 To perform a position accuracy calibration for well plates Step Note 1 Switch to the Diagnosis View of the Agilent ChemStation Software 2 Select Tests from the Diagnosis menu 3 Inthe Test Selection dialog box select Micro AFC G1364D from the drop down list Agilent 1200 Series Micro Collector Spotter User M anual Troubleshooting and Test Functions 4 To perform a position accuracy calibration for well plates continued Step Note 4 Select position accuracy calibration well plates and click on Start 5 Start the position accuracy calibration and Click on Explain to get detailed instructions follow the instructions on the screen and information 6 Align the capillary tip to the center of each See Figure 19 on page 101 The height corner well with the corresponding positioning calibration is not required for well plates and buttons therefore greyed out 7 Repeat Step 6 for all configured plates OO hl Adjust
48. e unplug the power cord The power supply still uses some power even if the power switch at the front panel is turned OFF WARNING To avoid personal injury keep fingers away from the needle area during micro collector spotter operation Do not attempt to insert or remove a vial or a plate when the needle is positioned po000g0 Fraction Collector 3 SSsSsssss Thermostat cable 3 e e l H l O00009 B000mA D CAN bus AC Power Figure6 Connections at the Rear of the Thermostatted Fraction Collector 26 Agilent 1200 Series Micro Collector Spotter User M anual Installing the Micro Collector Spotter 1 Micro Collector Spotter Trays Installing the Fraction Collector Trays 1 Press the front door latch holding button located at the front of the right side cover 2 Lift the front door 3 Adjust the top plate of the test tube tray for the correct tube height if required 4 Load the micro collector spotter tray with micro collector spotter well plates test tubes or vials as required 5 Slide the micro collector spotter tray into the micro collector spotter so that the rear of the tray is seated firmly against the rear of the tray area Installed trays are automatically detected and identified 6 Press the front of the micro collect
49. e warranty terms in the sep arate agreement shall control Technology Licenses The hardware and or software described in this document are furnished under a license and may be used or copied only in accor dance with the terms of such license Restricted Rights Legend If software is for use in the performance of a U S Government prime contract or subcon tract Software is delivered and licensed as Commercial computer software as defined in DFAR 252 227 7014 J une 1995 oras a commercial item as defined in FAR 2 101 a or as Restricted computer soft ware as defined in FAR 52 227 19 J une 1987 or any equivalent agency regulation or contract clause Use duplication or disclo sure of Software is subject to Agilent Tech nologies standard commercial license terms and non DOD Departments and Agencies of the U S Government will receive no greater than Restricted Rights as defined in FAR 52 227 19 c 1 2 J une 1987 U S Government users will receive no greater than Limited Rights as defined in FAR 52 227 14 J une 1987 or DFAR 252 227 7015 b 2 November 1995 as applicable in any technical data Safety Notices CAUTION A CAUTION notice denotes a haz ard It calls attention to an operat ing procedure practice or the like that if not correctly performed or adhered to could result in damage to the product or loss of important data Do not proceed beyond a CAUTION notice until the indica
50. ector Spotter User Manual 6 Parts and Materials List of Recommended Plates Table 12 Recommended Plates Item Description Select in ChemStation Volume ml Package Part Number 1 96 polypropylene well plate 96A gilent 0 5 10 5042 1386 2 96 polypropylene well plate 96A gilent 0 5 120 5042 1385 3 384 polypropylene well plate 384Agilent 0 1 30 5042 1388 4 96 polypropylene conical well plate 96EppendorfC 0 18 25 5042 8502 5 54 x 2ml vial plate 54VialPlate 15 6 G2255 68700 Only one type of well plates can be used at a time in one tray WARNING If you are using flammable solvents remove the plates from the micro collector spotter after turning it OFF You avoid the risk of building explosive gas mixtures in the instrument 130 Agilent 1200 Series Micro Collector Spotter User M anual Parts and Materials 6 MALDI Spotting Accessories Table 13 MALDI Plate Carriers Plate Type Part Number MALDI Plate Carrier for Agilent Technologies AP M ALDI Targets 5022 6543 MALDI Plate Carrier for Bruker AnchorChip 5022 6541 MALDI Plate Carrier for Bruker Prespotted AnchorChip 5022 6546 MALDI Plate Carrier for Applied Biosystems Targets standard format 5022 6542 MALDI Plate Carrier for Applied Biosystems Opti TOF MTP format 5023 0238 MALDI Plate Carrier for Micromass Targets 5022 6544 Table 14 Calibration Plates Plate Type Part Number Calibration Plate for Agilent Technologies AP M ALDI Targets 5023 0214
51. ellow waiting for the manual reset Before you reset the micro collector spotter the door has to be closed To reset the instrument click on the micro collector spotter icon in the graphical user interface of the ChemStation and start the Reset If the main pump is switched ON a dialog is displayed which reminds the user to switch OFF the pump Then the reset procedure is started without the risk of contaminating well plates or fractions During the initialization process the vertical and the horizontal position of the capillary tip is determined The micro collector spotter performs the capillary check by pressing the capillary tip onto the sensor pad After this capillary check a message is displayed if any further capillary alignment is required For more details about capillary alignment refer to Choosing and Installing the Micro Collector Spotter Capillary on page 38 Switching the micro collector spotter lamp The micro collector spotter includes a lamp to illuminate the fraction compartment The lamp is specifically required for the position accuracy calibration as described on page 100 and page 102 The operation of the lamp requires firmware revision A 05 09 or higher and ChemStation revision A 10 02 or higher To activate the lamp switch open the command line in the ChemStation and type microafclampon to switch the lamp ON microafclampoff to switch the lamp OFF Agilent 1200 Series Micro Collector Spotter User M anual
52. embly Micro Scale Item Description Part Number 1 Internal tray assembly micro scale includes items 2 6 G1364 63115 2 Waste tubing with fitting 61364 86711 3 Washer 3050 2204 4 Screw cap 9301 1379 5 Flap septum G1364 27107 6 Waste Container G1364 22301 O o ae e au 4 Figure 24 Internal Tray Assembly M icro Scale 136 Agilent 1200 Series M icro Collector Spotter User M anual Parts and Materials 6 Micro Collector Spotter Accessory Kit Table 19 Micro Collector Spotter Accessory Kit Contents G1364 68715 Description Quantity Part Number Hex key 2 0 mm 1 8710 2476 MALDI Spotting Adapter 1 61364 83205 Flap Septum 1 61364 27107 Waste Tubing 1 4 mm ID 1 61364 86711 Waste tubing 1 2 m 1 5062 2463 CAN cable 1m 1 5181 1519 Sticking clamp for corrugated waste tubing large 3 5065 9976 Sticking clamp for waste tubing small 3 5065 9976 ESD wrist strap 1 9300 1408 Fraction Collector Capillary Assembly 25 um ID 1 61364 87304 Fraction Collector Capillary Assembly 100 um ID 1 61364 87306 ZDV Union stainless steel 1 5022 2184 Strip with 10 MALDI Spotting Tips 1 61364 81701 Plastic Spacers 20 not orderable Plug for waste container 1 61364 26105 Reorder gives 5m t Reorder gives 10 clamps for corrugated waste tubing large and 10 clamps for waste tubing small Reorder gives 10 clamps for corrugated waste tubing large and 10 clamps for waste tubing small Agilent 12
53. enance functions 5 Maintenance and Simple Repairs This chapter contains instructions on maintenance and simple repair procedures 6 Parts and Materials This chapter contains selected illustrations and lists for identification of parts and materials that are required for maintenance or exchanged in simple repair procedures 7 Specifications This chapter contains performance specifications of the micro collector spotter Agilent 1200 Series Micro Collector Spotter User M anual 3 A Safety Information This appendix provides a safety summary Agilent 1200 Series Micro Collector Spotter User M anual Contents 1 Agilent 1200 Series Micro Collector Spotter User Manual Contents Installing the Micro Collector Spotter 9 Site Requirements 10 Power Consideration 10 PowerCords 11 BenchSpace 11 Environment 12 Unpacking the Fraction Collector 14 Damaged Packaging 14 Delivery Checklist 14 Accessory Kits 15 Optimizing the Stack Configuration 17 Installing the Micro Collector Spotter 19 Installing a Thermostatted Fraction Collector 23 Micro Collector Spotter Trays 27 Installing the Fraction Collector Trays 27 Numbering of vial well plate and spot positions 28 Configure Well plate Types 29 Transporting the Micro Collector Spotter 32 Configuration and Operation of the Micro Collector Spotter 33 Introduction 34 Initialization and Reset 37 Configuration of the Micro Collector Spotter 38 Choosing and Insta
54. ep the EMF flag set An exceeded limit will not stop a sequence or run information only to plan maintenance activities System Log Select F2 to open the System Log There the user can find information about errors or not ready conditions The System Log will be cleared when the micro collector spotter is switched off The System Log can be printed F6 or saved to a file on the PCMCIA card pressing the m key ice 0 O008GS Ready 10 25 05 16 55 2 10 25 05 16 55 15 10 25 05 16 53 27 Plate P2 detected 10 25 05 16 53 24 Plate P3 detected 10 25 05 16 53 20 86 Agilent 1200 Series Micro Collector Spotter User M anual Control Module Screens of the Fraction Collector 3 ErrorLog Select F3 to open the Error Log where all module errors are documented The Error Log is a lifetime logbook All entries will be stored on the mainboard of the instrument The Error Log can be printed F6 or saved to a file on the PCMCIA card pressing the m key ice 0 008 Ready Message nitialization failed 10 24 05 16 38 2 Touchpad malfunction 10 21 05 16 38 21 Thermostat contact lost 10 21 05 16 32 04 Thermostat contact lost 10 21 05 16 32 03 Thermostat contact lost 10 21 05 16 32 02 Maintenance Log Use the F4 key Maintenance Log to look for maintenance activities performed on the instrument A list of possible events are listed in the scroll screen e g Capillary replaced Select the activity you have performed and p
55. equency When defective Tools required None Parts required Internal Tray micro scale G1364 63115 Before beginning this procedure Position the transport unit of the micro collector spotter in the Home Position see Maintenance Functions on page 93 Remove all installed trays from the tray base Turn OFF the instrument 1 Locate the internal tray assembly with the rinse funnel and flow delay sensor in the bottom of the right front corner of the instrument 124 Agilent 1200 Series Micro Collector Spotter User M anual Maintenance and Simple Repairs 5 2 Remove the internal tray by pushing down the plastic holder that holds it in position underneath the metal latch 1 and sliding the tray to the left at the same time 2 3 Remove the corrugated waste tubing from the front of the instrument 1 and slide the internal tray s waste tubing to the rear of the outlet 2 the before sliding the tray out 4 Install the new tray by sliding it into position underneath the metal leash that holds it The waste tubing from the internal tray should be guided as shown 5 Make sure that the waste tubing is slid all the way through the outlet 1 Its end should be over the edge and below the level of the laboratory desk that the system stands on to avoid any back flow of solvent Re install the corrugated waste tubing 2
56. er Viewing your Results Report In order to create reports with a fraction table and tick marks the Specify Report dialog box the item Add Fraction Table and Tick Marks has to be checked Figure 13 Fraction collection report setup 58 Agilent 1200 Series Micro Collector Spotter User M anual Configuration and Operation of the Micro Collector Spotter 2 Online M atrix Delivery The Agilent 1200 Series micro collector spotter can be configured for online matrix delivery For this application the Online Matrix Kit G1364 68706 is required see Table 15 on page 131 This kit contains all accessories required to control a syringe pump and to connect the syringe to the LC flow path An overview is displayed in Figure 14 For low flow rates and for MALDI spotting with online matrix delivery we recommend using the plug to close the waste container instead of the waste tubing The plug G1364 26105 is included in the micro collector spotter accessory kit lt a UVDetector Pump Control Cable T Type Connector Syringe Pump ee Matrix Capillary pe tart MALDI Spotter Figure 14 Online M atrix Delivery Syringe Pump Control The Online Matrix Kit contains an External Contact Board BCD G1351 68701 to control the syringe pump In addition a cable to connect the KDS 200 syringe pump is provided 5181 1536 To connect other syringe pumps a Agilent 1200 Series Micro Collecto
57. erface to G1330A Thermostat CAN DC out for operation of Agilent approved external devices like valves Only one type of well plate or MALDI plate can be used at a time in one tray Agilent 1200 Series Micro Collector Spotter User M anual 141 7 Specifications Performance Specifications for the Micro Collector Spotter 142 Agilent 1200 Series Micro Collector Spotter User M anual Agilent 1200 Series Micro Collector Spotter User Manual A Safety Information Safety Information 144 Lithium Batteries Information 147 Radio Interference 148 SoundEmission 149 Solvent Information 150 Agilent Technologies on Internet 151 a Agilent Technologies 143 A Safety Information Safety Information The following general safety precautions must be observed during all phases of operation service and repair of this instrument Failure to comply with these precautions or with specific warnings elsewhere in this manual violates safety standards of design manufacture and intended use of the instrument Agilent Technologies assumes no liability for the customer s failure to comply with these requirements This is a Safety Class I instrument provided with terminal for protective earthing and has been manufactured and tested according to international safety standards If you are using flammable solvents remove the well plates from the tray when you turn off the sampler You avoid the risk of building explosive gas mixtures in
58. get 384 Opti TOF target M PT format 96 Opti TOF target M PT format Bruker 384 AnchorChip Sample 384 AnchorChip Lock 1536 AnchorChip 384 Prespotted AnchorChip Micromass Agilent 1200 Series Micro Collector Spotter User M anual 133 6 134 Parts and Materials Table 16 Supported Targets for M ALDI Spotting continued Note M ALDI target plates for Applied Biosystems Bruker and Micromass are not supplied by Agilent Technologies Dimensions for other targets can be configured in the ChemStation Software 96 Micromass Sample 96 Micromass Lock 84 Micromass Sample 84 Micromass Lock 84 Micromass Calib Agilent 1200 Series Micro Collector Spotter User M anual Parts and Materials 6 Transport Unit Assembly Table 17 Transport Unit Assembly Micro Scale Item Description Part Number 1 Transport unit assembly micro scale 61364 60020 includes items 2 4 2 Capillary carrier assembly micro scale G1364 60023 3 Capillary Guiding Assembly 61364 87303 4 Wellplate adapter 61364 23203 5a Fraction Collector Capillary Assembly 25 um ID G1364 87304 5b Fraction Collector Capillary Assembly 50 um ID G1364 87305 5c Fraction Collector Capillary Assembly 100 um ID 61364 87306 Figure 23 Transport Unit Assembly Micro Scale Agilent 1200 Series Micro Collector Spotter User Manual 135 6 Parts and Materials Internal Tray Assembly Table 18 Internal Tray Ass
59. gured for MALDI spotting The delay volume has been specified The MALDI adapter and the MALDI spotting tip have been installed V The Position Accuracy Calibration for MALDI Targets on page 102 has been executed before running the checkout V A MALDI plate carrier with supported MALDI target is installed as plate 1 in the 4 well plate tray Table 7 Method parameters for M ALDI spotting check out Agilent 1200 Series modules micro degasser capillary pump 20ul flow sensor or nanoflow pump e micro autosampler micro well plate sampler column compartment optional diode array detector multiple wavelength detector with 500nl or 80 nl flow cell optional micro collector spotter configured for M ALDI Spotting Column ZORBAX SB C18 5 um 150 x 0 5mm 5064 8256 for the capillary pump ZORBAX SB C18 3 5 um 150 x 100m 5065 9910 or similar column for the nanoflow pump Sample blank injection no sample required Flow 5 wl min capillary pump or 1 ul min nanoflow pump Stoptime 3 minutes capillary pump 6 minutes nanoflow pump Solvent A 30 HPLC grade water Agilent 1200 Series Micro Collector Spotter User M anual 65 2 66 Configuration and Operation of the Micro Collector Spotter Table 7 Method parameters for M ALDI spotting check out continued Solvent B 70 HPLC grade acetonitrile Injection volume blank injection Column temperature 25 0 C or ambient Fraction trigger mode Use Timeta
60. ick on the Start button in the graphical user interface of the ChemStation select Run Sequence from the RunControl menu e press F6 Agilent 1200 Series Micro Collector Spotter User M anual 55 2 Configuration and Operation of the Micro Collector Spotter Setting up a Micro Collector Spotter Method Online tick marks To display tick marks for fraction start and stop events in you Online Plot click on the Change button in your Online Plot window Then check Show fraction collection ticks in the Edit Signal Plot window Figure 11 Figure 11 Online Tick Marks 56 Agilent 1200 Series Micro Collector Spotter User M anual Configuration and Operation of the Micro Collector Spotter 2 Viewing your Results Viewing your Results Data Analysis In order to display the tick marks for the collected fractions or spots on the screen click on Signal options from the Graphics menu Then choose Separated in the Layout drop down menu To review your chromatograms file information and a fraction list select the Data Analysis view from the drop down menu and press the Fraction Task button as displayed in Figure 12 Purification Task Button Chromatogram with Fraction Tick EAEE a z Seeth EOIN eee GAIS 54 depte Fraction Table Figure 12 Data Analysis View Agilent 1200 Series M icro Collector Spotter User M anual 57 2 Configuration and Operation of the Micro Collector Spott
61. icted flow of the condensation into a waste container must be guaranteed M ake sure that the condensation drain tube is always above the liquid level in the container If the tube is located in liquid the condensed water cannot flow out of the tube and the outlet is blocked Any further condensation will then remain in the instrument This may damage the instruments electronics The micro collector spotter thermostat requires 25 cm 10 inch space on each for sufficient air circulation Install the LAN interface board in the micro collector spotter if required Remove the adhesive tape which covers the side and front doors Open the front door and remove the left side door Remove the transport protection foam nN Oo oO FP UW Install the corrugated waste tube in the plastic port at the front bottom center of the micro collector spotter and route down into a waste container Slide the waste tubing coming from the internal tray if present Agilent 1200 Series Micro Collector Spotter User M anual 23 1 24 Installing the Micro Collector Spotter through the plastic port and the corrugated waste tube see Figure 2 on page 20 Route the corrugated waste tubing into a waste container 8 Re install the left side door take care of the magnet at the back Ensure the side door is correctly installed its presence is sensed by a hall sensor a missing side door will result in a NOT READY state of the instrument 9 Place the
62. imetable If the LC system is used for analytical chromatography without fraction collection or MALDI spotting the micro collector spotter can be switched Off The spot size for M ALDI spotting is specified by the flow rate and the collection time If peak based trigger mode is selected for MALDI spotting the collection time and therefore the spot size is flexible In this case we recommend specifying a maximum fill volume per location as described in Auxiliary on page 50 48 Peak Detectors The Peak Detectors section in this screen comprises a list of all Peak Detectors connected to the system As working modes for each peak detector Threshold only Threshold Slope and Slope only are possible In the Threshold only mode the settings for Up Slope Down Slope and Upper Threshold in the subsequent columns are ignored Fraction collection is triggered whenever the detector signal exceeds the specified threshold value When the signal drops below the threshold value fraction collection is stopped In the Slope only mode fraction collection is triggered on the slopes of the detector signals Adequate values for Up Slope and Down Slope can be specified in the corresponding fields In the Threshold Slope mode fraction collection is triggered on the corresponding values for threshold and slope For more detailed information concerning working with threshold and slopes please refer to Agilent s Application Note Sophisticated peak based frac
63. in Posttime 0 00 min overrides HPLC system runtime settings Method screens Use Esc to return to the Analysis screen Use the F3 key Method to view the parameters in a method Use the F8 key Save As to save the method in the module s The PC Card key is only active when a PCMCIA card is inserted in the control module Module F1 and Method F2 give access to methods stored on the Agilent 1200 Series LC module or the PC Card Timetable F3 gives access to the timetable of a method if specified The PCM CIA card must have been inserted into the control module prior to the last start up in order to be recognized Setting 76 Agilent 1200 Series Micro Collector Spotter User M anual Control Module Screens of the Fraction Collector 3 After pressing F1 or F2 use the Right Left selection keys to switch between PC Card and Instrument window Use the Up Down selection keys to select the method Use F7 F8 Copy to copy a selected method from one window to the other one Use F6 Delete to delete a selected method Ts end of method list Delete Press the m key to open the context sensitive menu for printing the method directory Date Time User Comment 05 25 00 14 52 end of method list Agilent 1200 Series Micro Collector Spotter User M anual 77 3 Control Module Screens of the Fraction Collector sequence In the sequence
64. ing a with a 1 6 mm OD capillary and flush the waste tubing and the waste port thoroughly In order to avoid flooding of the fraction collector a tissue should be placed besides the internal tray close to the spill Assemble the cleaned waste tubing Install the internal tray in the fraction collector as shown in step 4 on page 125 IMPORTANT Make sure that the waste tubing is guided correctly through the waste tubing channel to the waste tubing outlet as shown in step 4 and step 5 on page 125 Agilent 1200 Series Micro Collector Spotter User Manual 111 5 Maintenance and Simple Repairs Exchanging the micro collector spotter lamp assembly Frequency When lamp is defective or not required in the micro collector spotter Tools required None Parts Required Micro collector spotter lamp assembly G1364 60014 WARNING To avoid personal injury keep fingers away from the needle area during micro collector spotter operation WARNING Explicitly follow the described installation procedures to maximize the lifetime of the fraction collector capillary and to avoid potential spills or fraction losses Regularly inspect the capillary and exchange it if itis worn out or shows visible signs of damage 112 Agilent 1200 Series Micro Collector Spotter User M anual Maintenance and Simple Repairs 5 Before beginning this procedure Position the transport unit of the micro collector spotter in the Home Posit
65. ing side door will result in a NOT READY state of the instrument 7 Place the micro collector spotter in the stack or on the bench in all horizontal position 8 Ensure the power switch at the front of the micro collector spotter is OFF Connect the power cable to the power connector at the rear of the micro collector spotter 10 Connect the CAN cable to the other Agilent 1200 Series LC modules 11 If an Agilent ChemStation is the controller it must be connected to the LAN interface should be installed to the detector 12 Connect the APG remote cable optional for non Agilent 1200 Series instruments 13 Install the tray that has been delivered for your micro collector spotter 20 Agilent 1200 Series Micro Collector Spotter User M anual Installing the Micro Collector Spotter 1 The micro collector spotter G1364D is shipped with 4 x well plate full tray All other trays have to be ordered separately 14 Turn ON power by pushing the button at the lower left hand side of the micro collector spotter 15 The exhaust fan will turn ON and remove potential solvent vapor from the inside of the instrument After 2 minutes close the front door Then the micro collector spotter will start the hardware initialization process At the end of this process the status LED should be green Agilent 1200 Series Micro Collector Spotter User M anual 21 1 Installing the Micro Collector Spotter PEPEE E J D le ontrol of Thermostat O THERMO
66. ion see Maintenance Functions on page 93 Remove all installed trays Position the transport unit of the micro collector spotter in the Change Parts Position see Maintenance Functions on page 93 Turn OFF the instrument Unscrew the micro collector spotter capillary from the flow cell of the detector Remove the fraction collector spotter capillary as described in Replacing Micro Collector Spotter Capillary Assembly on page 117 1 Unplug the lamp power cable from the connector at the front of the z arm 2 To disassemble the lamp carefully bend up the two metal sheets that hold the LEDs Then push up the lamp assembly 3 Un clip the cable connector which is located between the plastic clips of the capillary guiding assembly eee of the Trlr N R A Nad Agilent 1200 Series Micro Collector Spotter User Manual 5 Maintenance and Simple Repairs 4 Disconnect the lamp assembly from the extension cable at the cable connector and pull the cable through the guiding hole in the carrier assembly extension cable 5 To install the new lamp assembly unplug the connector as described in step 4 and slide the cable from the top through the hole in the carrier assembly guiding hole 6 Carefully bend up the two metal sheets of the lamp assembly that hold the LEDs Then slide the assembly onto the spotting adapter that the t shape
67. ions Agilent 1200 Series Micro Collector Spotter User M anual 31 1 Installing the Micro Collector Spotter Transporting the Micro Collector Spotter 32 When moving the Agilent 1200 Series micro collector spotter inside the laboratory no special precautions are needed However if the micro collector spotter needs to be shipped to another location via carrier ensure V The transport assembly is in the park position Use the ChemStation or the Control Module for this command To move the arm to the park position 1 Switch to the Diagnosis view of the ChemStation and select Fraction Collector gt Maintenance Positions from the Maintenance menu 2 In the upcoming dialog box click Park Arm V The vial tray and the sample transport mechanism is secured with the transport protection foam Agilent 1200 Series Micro Collector Spotter User M anual Agilent 1200 Series Micro Collector Spotter User Manual 2 Configuration and Operation of the Micro Collector Spotter Introduction 34 Initialization and Reset 37 Configuration of the Micro Collector Spotter 38 Setting up a Micro Collector Spotter Method 47 Viewing your Results 57 Check out procedures 63 aa Agilent Technologies 33 2 Configuration and Operation of the Micro Collector Spotter Introduction The Agilent 1200 Series micro collector spotter G1364D is an instrument for micro scale fraction collection or MALDI Spotting with the Agilent 1200 Series
68. ip After the maintenance or repair task has been finished click End to reset the micro collector spotter and move the arm to the waste port Home Position This maintenance function moves the arm up and to the right rear for better access and exchange of the trays Park Arm This maintenance position moves the arm to the park position at the upper rear left side of the tray for transporting or shipping the micro collector spotter 94 Agilent 1200 Series Micro Collector Spotter User M anual Troubleshooting and Test Functions 4 Transport Unit Self Alignment The transport unit alignment is required to compensate for larger deviations in positioning the needle carrier assembly This might be necassary after disassembling the system or when you exchange the transport unit the needle carrier assembly or the MTP main board This function is in the diagnose screen of the ChemStation or the Control Module WARNING The sample transport self alignment requires the 4 well plate tray G1364 84511 or G1364 84521 All well plates MUST be removed If the Transport Unit Self Alignment is started with well plates on the tray the alignment procedure is aborted WITHOUT error message When is a Transport Unit Self Alignment Necessary The sample transport self alignment is required after disassembling the module or when you exchange e The transport unit e The needle capillary carrier assembly e The MTP main board How
69. itable site environment is important to ensure optimum performance of the micro collector spotter Power Consideration The micro collector spotter power supply has wide ranging capability see Table 1 on page 12 Consequently there is no voltage selector in the rear of the micro collector spotter There are also no externally accessible fuses as automatic electronic fuses are integrated in the power supply The thermostatted micro collector spotter comprises two modules the micro collector spotter G1364D and the thermostat G1330B Both modules have a separate power supply and a power plug for the line connections The two modules are connected by a control cable and both are turned on by the micro collector spotter module The thermostat power supply has two externally accessible fuses To disconnect the micro collector spotter from line power unplug the power cord The power supply still uses some power even if the power switch on the front panel is turned off To disconnect the thermostatted micro collector spotter from line power unplug the power cord from the micro collector spotter and the thermostat The power supplies still use some power even if the power switch on the front panel is turned off Please make sure that itis always possible to access the power plug Shock hazard or damage of your instrumentation can result if the devices are connected to a line voltage higher than specified 10 Agilent 1200 Se
70. l plate tray mucas 0 O08 Ready Plate Formats 192AppliedBiosys 24WhatmanUniPlat 384AnchorChipLo 384AnchorChipSa 384ApogentC Active Plates P4 384Agilent P3 384Agilent P2 384Agilent F1 384Agilent O 384 4x 96 Select highlight a tray configuration with the help of the selection keys F7 F8 Add Remove allows you to move configurations from available Plate Agilent 1200 Series Micro Collector Spotter User M anual 81 3 Control Module Screens of the Fraction Collector Configure fraction Configure fraction delays 82 Formats to the Active Plates and vice versa Press F2 Delete Def to delete a user defined definition Press F3 Rotate Plate to rotate the selected plate Use F1 Definition to edit the user defined definition of a well plate Changes must be acknowledged with F6 Done Only one type of well plate or M ALDI target can be selected at the same time F3 Fraction in the main Configuration screen allows you to access the Delay volume definition screen to specify Filling Order and Reserved or Needle Movement ce 60 OORRIE Ready 2 Filling Order Reserved Locations 4 Needle Move Interfaces Delays in the Fraction Configuration screen allows you to configure the delay volume between each connected detector and the micro collector spotter Edit F8 allows to enter new values for the selected detector Changes
71. lling the Micro Collector Spotter Capillary 38 Configuration of the ChemStation 42 Setting up a Micro Collector Spotter Method 47 Fraction Preview 51 Contents Starting your run with fraction collection or M ALDI spotting 53 Assignment of StartLocation 54 Online tick marks 56 Viewing your Results 57 Data Analysis 57 Report 58 Online M atrix Delivery 59 Check out procedures 63 Check out procedure for micro fraction collection 63 Check out procedure for M ALDI spotting 65 Control M odule Screens of the Fraction Collector 67 Major keys on the Agilent 1200 Series Control Module 68 Screens available from the Analysis view 69 Screens available from the System view 80 Troubleshooting and Test Functions 89 Status Indicators 90 Power Supply Indicator 90 Instrument Status Indicator 91 Deactivating the door lock sensor 92 Maintenance Functions 93 Transport Unit Self Alignment 95 Arm Length Calibration 97 Position Accuracy Calibration for 384 Well Plates 100 Position Accuracy Calibration for M ALDI Targets 102 Single Step Commands 105 Agilent 1200 Series Micro Collector Spotter User M anual Contents 5 Maintenance and Simple Repairs 107 Introduction into Repairing the Micro Collector Spotter 108 Simple Repairs 108 Cleaning the Micro Collector Spotter 108 Cleaning the waste port 109 Exchanging the micro collector spotter lamp assembly 112 Simple Repairs Procedures 116 Replacing Micro Collector Spotter Capillary As
72. m fromthe topofthe upto 45mm plate to the bottom of the well If you use well plates with inlets caps and septa you have to specify the distance from the top of the septa to the bottom of the inlets 30 Agilent 1200 Series Micro Collector Spotter User M anual Installing the Micro Collector Spotter 1 Table7 Configuring Well plate Types continued Location Description Definition Limits Well X size Size of the well in x direction Plate min 3 7mm length If you use well plates with min 3 0 mm with inlets caps and septa youhave to position accuracy specify the x size of the septa alignment micro scale Well Y size Size of the well in y direction Plate down to 3 7mm width If you use well plates with min 3 0 mm with inlets caps and septa youhaveto position accuracy specify the y size of the septa alignment micro scale Bottom size For round wells the relative of the 1 0 cylindrical well top and bottom of the well 0 0 conical well Square Click in the field to specify whether Yes rectangular the well is rectangular or round No round oval Is well plate Click in this field to specify if this is Yes well plate or a well plate or not Relevant for continuous flow operation MALDI Target No Vial Tray or Eppendorf tray The distances need to be measured with high precision It is recommended to use a caliper If possible contact the vendor of non predefined well plates to obtain these dimens
73. mber P1 P4 row is A to H and column 1 12 for 96 well plates or the Agilent AP MALDI plate for different well plates or MALDI plates refer to the description from the plate manufacturer Note the orientation of the M ALDI plate The starting position A1 is always in the back left corner of the plate x Operstoe Name fio Chemeat Data Fila C Pretia Comster Manoni Hennesee oErAUL TO Sebdenctory PRT Ome Path C MPCHEMSIWDATA Siompte Name Sample 61 Sampie Anot mper ff Siem P tive ft 8 Figure9 Sample Info dialog box All settings made in the Sequence Table overrule the settings chosen in the Sequence Parameters screen In addition to the exact position for fraction collection start in the Sequence Table it is also possible to specify Next Plate and Next Location Then the fraction collection is started at the next free plate or at the next free location If no fraction start location has been specified the fraction collection starts at first empty location on the well plate or MALDI target Agilent 1200 Series Micro Collector Spotter User M anual Configuration and Operation of the Micro Collector Spotter Setting up a Micro Collector Spotter M ethod Figure 10 Start Location in Sequence Parameters dialog box To start a single run e click on the Start button in the graphical user interface of the ChemStation e select Run Method from the RunControl menu e press F5 To start the sequence e cl
74. micro collector spotter on top of the thermostat Make sure that the micro collector spotter is correctly engaged in the thermostat locks 10 Remove the plastic cover from the tray base place the air channel adapter 1 into the micro collector spotter tray base Make sure the adapter is fully pressed down This assures that the cold airstream from the thermostat is correctly guided to the tray area of the micro collector spotter Place the plug channel 2 on top of the air channel adapter Both devices must be installed correctly to assure proper operation of the instrument Figure4 Installation of Thermostat and Fraction Collector 11 The Agilent 1200 Series micro collector spotter is delivered with a pre installed tray compartment divider This divider should only be used with the thermostatted micro collector spotter if a half or std tray is installed into the left and center positions of the instrument If any tray is installed to the right side of the instrument remove the tray compartment divider The tray compartment divider optimizes the cooling performance of the instrument if only the left and center position of the micro collector spotter are in use Agilent 1200 Series Micro Collector Spotter User M anual Installing the Micro Collector Spotter 1 Figure5 Removing the Tray Compartment Divider 12 Install the tray you have ordered for your micro collector spotter 13 Ensure the power switch on the front of
75. must be acknowledged with F6 Done Detector Agilent 1200 Series M icro Collector Spotter User M anual Configure fraction filling order Configure reserved locations Configure Needle Movement Control Module Screens of the Fraction Collector 3 Filling Order in the Fraction Configuration screen allows you to configure the filling order of the trays in the micro collector spotter Edit F8 allows to enter new values for the selected detector Changes must be acknowledged with F6 Done Nh O Shortest Way by Column Column by Shortest Way O Column O by Row Reserved Locations allows you to specify reserved locations by Rows Columns or Single Locations that are not used filled during the collection Changes must be acknowledged with Done F6 Reserve ORow s sans CI columnis Localorns Single E Losations s The Needle Movement allows you to specify the position of the needle capillary tip during fraction collection With into location the capillary tip moves to the specified depth in the well With above location the capillary tip stays at the specified distance above the well For micro fraction collection or MALDI spotting we recommend to use the contact control mode In this mode the capillary tip moves to the specified distance above the bottom of the well or the MALDI target When the fraction is collected the needle will move upwards while staying in cont
76. o interference limits are still met within the premises 148 Agilent 1200 Series M icro Collector Spotter User M anual Safety Information A Sound Emission M anufacturer s Declaration This statement is provided to comply with the requirements of the German Sound Emission Directive of 18 January 1991 This product has a sound pressure emission at the operator position lt 70 dB e Sound Pressure Lp lt 70 dB A e At Operator Position e Normal Operation e According to ISO 7779 1988 EN 27779 1991 Type Test Agilent 1200 Series Micro Collector Spotter User M anual 149 A Safety Information Solvent Information Observe the following recommendations on the use of solvents This instrument should only be used with solvents that have an ignition temperature higher than 200 C 150 Solvents Brown glass ware can avoid growth of algae Always filter solvents small particles can permanently block the capillaries Avoid the use of the following steel corrosive solvents Solutions of alkali halides and their respective acids for example lithium iodide potassium chloride and so on High concentrations of inorganic acids like nitric acid sulfuric acid especially at higher temperatures replace if your chromatography method allows by phosphoric acid or phosphate buffer which are less corrosive against stainless steel Halogenated solvents or mixtures which form radicals and or acids for example
77. ode Call your local service provider for assistance upon observing this error condition A flashing red status indicator indicates an during the startup procedure of the module Call your local service provider for assistance upon observing this error condition Agilent 1200 Series M icro Collector Spotter User M anual 91 4 Troubleshooting and Test Functions Deactivating the door lock sensor The Agilent 1200 Series micro collector spotter G1364D allows the user to operate the instrument with an open front door In order to use this option the door lock sensor has to be disabled Type the following command into the ChemStation Command Line doorunlock The door lock sensor should only be disabled for special applications It should be activated again if this operation mode is no longer necessary To activate the door lock sensor again type doorlock If the door lock sensor is disabled the instrument will stay in the READY status while the door is open However the fraction collector will always startup in the default mode door lock on after it has been power cycled If the door lock sensor is disabled the fraction collector arm will move while the front door is open To avoid personal injury keep fingers away from the needle area during fraction collector operation All users must be informed about this potential risk 92 Agilent 1200 Series Micro Collector Spotter User M anual Troubleshooting and Test Function
78. ommended Plates on page 130 you may configure a custom plate Measure the exact dimensions of the plate as marked below and enter the values in the plate configuration table of the ChemStation or the Control Module O J Figure7 Well plate Dimensions straight Figure8 Well plate Dimensions staggered Agilent 1200 Series M icro Collector Spotter User M anual 29 1 Installing the Micro Collector Spotter Table 7 Configuring Well plate Types Location Description Definition Limits Rows Number of rows on the plate up to 16 Columns Number of columns on the plate up to 24 Volume Volume in ul of a sample vessel A Row distance Distance in mm between the center of two rows B Column distance Distance in mm between the center of two columns C Plate length X size in mm at the bottom of the 127 75 0 25 mm plate SBS Standard D Plate width Y size in mm at the bottom ofthe 85 50 0 25 mm plate SBS Standard E Plate height Size in mm from the bottom to the up to 47 mm top of the plate If you use well plates with inlets caps and septa you have to specify the distance from the bottom to the top of the caps F Column offset Distance in mm from the left edge bottom to the center of the first hole A1 G Row offset Distance in mm from the back edge bottom to the center of the first hole A1 H Column shift Offset in mm to Y when the rows are not straight but staggered J Well depth Distance in m
79. on see Maintenance Functions on page 93 Remove all installed trays from the tray base Position the transport unit of the micro collector spotter in the Exchange Parts Position see Maintenance Functions on page 93 Turn OFF the instrument IMPORTANT Do not adjust the capillary while the clamp nut is tightly mounted on the capillary guide 1 Remove the wellplate adapter and the clamp nut Then pull the capillary from the clip at the bottom of the carrier assembly as well as the capillary guiding assembly 120 Agilent 1200 Series Micro Collector Spotter User M anual Maintenance and Simple Repairs 5 2 Press the yellow clips of the capillary guiding assembly together and pull the assembly towards the rear of the capillary carrier assembly 3 Slide the new capillary guiding assembly into the holder of the capillary carrier assembly M ake sure to push it all the way to the front adapter capillary installed 4 Reinstall the Capillary the clamp nut and the wellplate IM PORTANT Do not tighten the clamp nut without a ae On completion of this procedure Re install the tray s in the tray base Start the instrument Close the front cover Agilent 1200 Series Micro Collector Spotter User Manual 121 5 Maintenance and Simple Repairs Exchanging the Flap Septum and
80. on collector arm to the Change Parts position see Maintenance Functions on page 93 Turn the well plate adapter 90 degrees and remove it from the capillary carrier arm Slide the spotting adapter onto the capillary carrier arm and turn it 90 degrees to fixate it The capillary length should be aligned as displayed in Figure 4 Attach the MALDI spotting tip to the capillary The MALDI spotting tip should be aligned to from a flat surface with the capillary tip For correct alignment leave the spotting tip in the plastic strip and push the spotting tip onto the capillary Agilent 1200 Series Micro Collector Spotter User M anual Configuration and Operation of the Micro Collector Spotter 2 Configuration of the Micro Collector Spotter Spotting Adapter 5 7mm ns Fraction Collector Capillary MALDI Spotting Tip Figure4 Capillary Alignment for M ALDI Spotting Agilent 1200 Series Micro Collector Spotter User M anual 41 2 Configuration and Operation of the Micro Collector Spotter Micro Collector Spotter Configuration of th Configuration of the ChemStation To setup or change the configuration parameters of your fraction collector select More Fraction Collector gt Configuration from the Instrument menu or right click on the fraction collector icon in the graphical user interface In the Fraction Collector Configuration dialog box Figure 5 the configuration of the Trays the Fra
81. or spotter tray down to secure the tray in the micro collector spotter 7 Close the front door If the tray pops out of position the air channel adapter is not correctly inserted Before starting a run the instrument has to be correctly configured in the user interface Agilent 1200 Series Micro Collector Spotter User M anual 27 1 Installing the Micro Collector Spotter Numbering of vial well plate and spot positions With the 4 plates full tray Plate in the left front position P1 Plate in the left back position P2 Plate in the right front position P3 Plate in the right back position P4 Vessel Al A2 B1 B2 With the 2 plates 10 x 2ml vials or 10 funnels std trays Plate in the front position P1 Plate in the back position P2 Vessel Al A2 B1 B23 Vials funnels 1 10 With the 100 vials std tray Vial 1 100 With the half trays Left hand 40 position tray 1 40 Center 40 position tray 101 140 Right hand 40 position tray 201 240 or Left hand 15 position tray 1 15 Center 15 position tray 101 115 Right hand 15 position tray 201 215 WARNING Do not use closing mats The micro collector spotter has no needle therefore the capillary will be bend or clogged after several injections 28 Agilent 1200 Series Micro Collector Spotter User M anual Installing the Micro Collector Spotter 1 Configure Well plate Types If the plate you are using is not found on the List of Rec
82. otective clothing as described in the material handling and safety data sheet supplied by the solvent vendor especially when toxic or hazardous solvents are used WARNING To avoid personal injury keep fingers away from the needle area during micro collector spotter operation Do not attempt to insert or remove a vial or a plate when the needle is positioned Install the LAN interface board in the micro collector spotter if required Remove the adhesive tape which covers the side and front doors Open the front door and remove the left side door Remove the transport protection foam ur WN Pe Install the corrugated waste tube in the plastic port at the front bottom center of the instrument Slide the waste tubing coming from the internal tray if present through the plastic port and the corrugated waste tube see Figure 2 on page 20 Route the corrugated waste tubing into a waste container Agilent 1200 Series Micro Collector Spotter User M anual 19 1 Installing the Micro Collector Spotter wal a al Ay Figure 2 Installing the Corrugated Waste Tubing in the Plastic Port 6 Re install the left side door take care of the magnet at the back Ensure the side door is correctly installed its presence is sensed by a hall sensor a miss
83. ows the configuration recommended for the micro collector spotter within an Agilent 1200 Series capillary or nanoflow LC system Agilent 1200 Series Micro Collector Spotter User M anual 17 1 Installing the Micro Collector Spotter Optimizing the Stack Configuration Solvent Compartment Micro Degasser E 1 Column Compartment re ggm umn Lomparment Capillary Nano _ Pump UV Vis detector gt 2D Micro 0 N Micro Well Plate Sampler i collector spotter Sampler Thermostat z Fraction Collector Thermostat Figure1 Recommend Stack Configuration for micro collector spotter in the capillary or nanoflow system NOTE The micro collector spotter should never be positioned on top of a module that generates heat which can lead to an unwanted evaporation of fractions in the micro collector spotter e g Agilent 1200 Series thermostatted column compartment G1316A B or Agilent 1200 Series diode array detectors G1315B C 18 Agilent 1200 Series Micro Collector Spotter User M anual Installing the Micro Collector Spotter 1 Installing the Micro Collector Spotter Preparation Locate bench space Provide power connections Unpack the micro collector spotter Parts required Fraction Collector Power cord Chemstation and or Control M odule G1323B WARNING When opening capillary or tube fittings solvents may leak out Please observe appropriate safety procedures for example goggles safety gloves and pr
84. p should be aligned just above the M ALDI Target leaving enough space for sheet of paper to fit between the capillary tip and the target 8 Repeat Step 6 7 for all configured plates Agilent 1200 Series Micro Collector Spotter User M anual 103 4 Troubleshooting and Test Functions Position Accuracy Calibration for M ALDI Targets xy position alignment buttons height z position alignment buttons Only available for MALDI targets Corrective Values relative to the previous alignment Figure 20 Position Accuracy Calibration Interface for M ALDI Targets 104 Agilent 1200 Series Micro Collector Spotter User M anual Troubleshooting and Test Functions 4 Single Step Commands Some movements of the fraction collection or spotting sequence can be done under manual control This is useful during troubleshooting where close observation of each of the fraction collection steps is required to confirm a specific failure mode or verify successful completion of a repair Each step command actually consists of a series of individual commands which move the micro collector spotter components to predefined positions enabling the specific step to be done In the ChemStation the step commands can be selected from the Test Selection Box see Figure 21 in the Diagnosis display In the Control Module the step commands can be accessed from the pull down menu in the micro collector spotter Test Automatic Fraction Collector Step
85. r Spotter User M anual 59 2 60 Configuration and Operation of the Micro Collector Spotter general purpose control cable 18594 60520 can be ordered and configured The BCD board can be installed in any Agilent 1200 Series LC module For installation details refer to the corresponding service manual The pump control can easily be setup in the ChemStation as displayed in Figure 15 To open the dialog box select the module where the external contact board is installed and click on the icon in the graphical user interface Then select Contacts The default setting should be set as open i e the syringe pump is switched off In the example that is shown in Figure 15 the syringe pump is switched off by default Using the Timetable the matrix flow can be switched as required during the run We recommend switching the syringe pump on at least 10 minutes before the start of the LC run in order to prime the matrix capillary and ensure that the matrix is added to the LC flow immediately after the run is started Contact 1 is open syringe pump is switched off Contact 1 is closed syringe pump is switched on CI x Contact 1 Contact 2 Contact 3 Comact 4 open C open open apen closed closed closed closed Teevtable Time mn C1 c2 c3 c4 Insert 1 O00 closed Append 2L Ol open Pe dood mae ul Copy Paste Figure 15 Syringe Pump Control through External Contacts Capillaries and Fittings The Online Matrix Kit con
86. r to ensure reproducible fraction collection and MALDI spotting without cross contamination and an accurate control of the collected volumes the liquid contact control mode as illustrated in Figure 2 on page 36 has been developed During the entire collection process the instrument ensures that the droplet at the capillary tip is in constant contact with the surface of the well the MALDI plate or the liquid being collected At the start of fraction collection the droplet is delivered at the bottom of the well or the MALDI plate While the well is filled the capillary tip moves continuously upwards keeping in constant contact with the liquid surface At the end of fraction collection the capillary tip moves sharply upwards in order to cut the contact between the liquid and the capillary During the collection the flow rate and the geometry of the well are taken into account to calculate the current fill level Agilent 1200 Series Micro Collector Spotter User M anual 35 2 Configuration and Operation of the Micro Collector Spotter Introduction Figure2 Liquid Contact Control 36 Agilent 1200 Series Micro Collector Spotter User M anual Configuration and Operation of the Micro Collector Spotter 2 Initialization and Reset After power on the instrument delays the initialization for approximately two minutes to vent the fraction compartment Then the door lock is activated and the micro collector spotter remains in the not ready condition y
87. rds IEC CSA UL EN Installation Category II Pollution Degree 2 For indoor use only Agilent 1200 Series Micro Collector Spotter User Manual 13 1 Installing the Micro Collector Spotter Unpacking the Fraction Collector CAUTION If you need to ship the micro collector spotter at a later date always use the shipping protection foam parts see Transporting the Micro Collector Spotter on page 32 Damaged Packaging Upon receipt of your micro collector spotter inspect the shipping containers for any signs of damage If the containers or cushioning material are damaged keep them until the contents have been checked for completeness and the micro collector spotter has been mechanically and electrically checked If the shipping container or cushioning material is damaged notify the carrier and keep the shipping material for the carrier s inspection If there are signs of damage to the micro collector spotter please do not attempt to CAUTION i install the micro collector spotter Delivery Checklist Ensure all parts and materials have been delivered with the micro collector spotter For this compare the shipment content with the checklist included in each instrument box Please report missing or damaged parts to your local Agilent Technologies sales and service office The Agilent 1200 Series micro collector spotter is available as G1364D micro collector spotter designed for flow rates below 100 ul and for use
88. ress Done to create an entry in the logbook Editing the add line with the help of the alphanumeric keys allows you to enter your own maintenance function For troubleshooting reasons this Logbook can be printed or saved to a file on the PCMCIA card pressing the m key Changes must be acknowledged with Done F6 Records Message Date Time capillary G1315 68708 replaced cl 05 23 05 15 16 49 cover replaced on 10 may 05 AT 05 10 05 13 01 41 and of logbook Agilent 1200 Series Micro Collector Spotter User M anual 87 3 Control Module Screens of the Fraction Collector Screens available from the System view 88 Agilent 1200 Series Micro Collector Spotter User M anual Agilent 1200 Series Micro Collector Spotter User Manual 4 Troubleshooting and Test Functions Status Indicators 90 Deactivating the doorlocksensor 92 Maintenance Functions 93 Transport Unit Self Alignment 95 Arm Length Calibration 97 Position Accuracy Calibration for 384 Well Plates 100 Position Accuracy Calibration for M ALDI Targets 102 Single Step Commands 105 a Agilent Technologies 89 4 Troubleshooting and Test Functions Status Indicators Two status indicators are located at the front panel of the micro collector spotter The lower left indicates the power supply status the upper right indicates the micro collector spotter status Status indicator green yellow red Line power switch with green light
89. ries Micro Collector Spotter User M anual Installing the Micro Collector Spotter 1 Power Cords Your micro collector spotter is delivered with a power cord which matches the wall socket of your particular country or region The plug on the power cord which connects to the rear of the instrument is identical for all types of power cord Never operate your instrumentation from a power outlet that has no ground connection Never use a power cord other than the power cord designed for your region Never use cables other than the ones supplied by Agilent Technologies to ensure proper functionality and compliance with safety or EMC regulations Bench Space The micro collector spotter dimensions and weight see Table 1 on page 12 allow the instrument to be placed on almost any laboratory bench The instrument requires an additional 2 5 cm 1 0 inch of space on either side and approximately 8 cm 3 1 inches at the rear for the circulation of air and room for electrical connections Ensure the micro collector spotter is installed in a horizontal position The thermostatted micro collector spotter dimensions and weight see Table 2 on page 13 allow the instrument to be placed on almost any laboratory bench The instrument requires an additional 25 cm 10 inches of space on either side for the circulation of air and approximately 8 cm 3 1 inches at the rear for electrical connections Ensure the micro collector spotter is installed
90. rnative micro collector spotter capillary 50um ID pre G1364 87305 500 50 1000 mounted micro collector spotter capillary 100um ID G1364 87306 500 100 4000 alternative een Due to the tolerance for the capillary ID small adjustments in delay volume might be necessary in order to optimize the performance of your system Collection Order The Collection Order describes capillary movement during fraction collection Four different strategies are possible row by row and column by column either in one direction or in two directions Collection Mode Selection of the Collection Mode is not available for the micro collector spotter and therefore greyed out in this configuration The micro collector spotter always operates in the continuous flow mode Needle Movement Intolocation In the into location mode the capillary tip moves into the well to the specified Depth in mm Above location In the above location mode the capillary tip stays at the specified Distance in mm above the well during fraction collection 44 Agilent 1200 Series Micro Collector Spotter User M anual Configuration and Operation of the Micro Collector Spotter 2 Contact Control In this mode the capillary tip moves down until is reaches the specified distance between capillary tip and well bottom This ensures that the forming droplet is in contact with the well bottom During the continuing filling process the capillary tip moves upwards while sta
91. s Needle Up Move to Location Vial Needle into Vessel Figure 21 Fraction Collector Step Commands Agilent 1200 Series Micro Collector Spotter User Manual 105 4 Troubleshooting and Test Functions Table9 Step Commands Step Action Comments Needle Up Lifts the needle arm to the upper position Needle into vessel Lowers the needle into the specified vessel Needle to rinse flush Moves the needle to the rinse flush port port 106 Agilent 1200 Series Micro Collector Spotter User M anual Agilent 1200 Series Micro Collector Spotter User Manual 5 Maintenance and Simple Repairs Introduction into Repairing the Micro Collector Spotter 108 Simple Repairs 108 Cleaning the Micro Collector Spotter 108 Cleaning the waste port 109 Exchanging the micro collector spotter lamp assembly 112 Replacing Micro Collector Spotter Capillary Assembly 117 Exchanging the Capillary Guiding Assembly 120 Exchanging the Flap Septum and the Waste Tubing 122 Exchanging the Internal Tray 124 ot Agilent Technologies 107 5 Maintenance and Simple Repairs Introduction into Repairing the Micro Collector Spotter Simple Repairs The micro collector spotter is designed for easy repair The most frequent repairs and maintenance tasks such as changing the capillary the flap septum or the waste tubing can be done from the front of the instrument with the instrument in place in the system stack These repairs are described in
92. s 4 Maintenance Functions Some maintenance procedures require the needle arm and needle carrier to move to specific positions to enable easy access to these components The maintenance functions move these assemblies into the appropriate maintenance position All trays have to be removed before starting the Maintenance dialog In the ChemStation the micro collector spotter maintenance positions can be selected from the Maintenance menu in the Diagnosis view see Figure 17 In the Control Module the functions can be selected in the Test screens of the micro collector spotter Fraction collector Maintenance Positions Change Parts End Move Arm Home Park Arm for Transport eS Figure 17 Fraction Collector M aintenance Positions dialog box Agilent 1200 Series Micro Collector Spotter User M anual 93 4 Troubleshooting and Test Functions Change Parts If you click Start the transport unit will move upwards the needle carrier assembly will move to the front center and then turn off the rotary motor to allow free rotation of the arm This position enables easy access e to change the capillary guiding assembly e to change the micro collector spotter capillary e to change the capillary carrier assembly e to change the micro collector spotter lamp assembly e to replace the well plate adapter with the spotting adapter or vice versa e to align the capillary length and install the spotting t
93. sembly 117 Exchanging the Capillary Guiding Assembly 120 Exchanging the Flap Septum and the Waste Tubing 122 Exchanging the Internal Tray 124 6 PartsandMaterials 127 Supported Trays for the Micro Collector Spotter 128 List of Recommended Plates 130 M ALDI Spotting Accessories 131 Supported M ALDI Targets 133 Transport Unit Assembly 135 Internal Tray Assembly 136 Micro Collector Spotter Accessory Kit 137 7 Specifications 139 Performance Specifications for the Micro Collector Spotter 140 A Safety Information 143 Safety Information 144 General 144 Operation 145 Safety Symbols 146 Agilent 1200 Series M icro Collector Spotter User M anual 7 Contents Lithium Batteries Information 147 Radio Interference 148 Test andMeasurement 148 Sound Emission 149 Manufacturer s Declaration 149 Solvent Information 150 Solvents 150 Agilent Technologies on Internet 151 8 Agilent 1200 Series Micro Collector Spotter User M anual Agilent 1200 Series Micro Collector Spotter User Manual 1 Installing the Micro Collector Spotter Site Requirements 10 Unpacking the Fraction Collector 14 Optimizing the Stack Configuration 17 Installing the Micro Collector Spotter 19 Installing a Thermostatted Fraction Collector 23 Micro Collector Spotter Trays 27 Configure Well plate Types 29 Transporting the Micro Collector Spotter 32 a Agilent Technologies 1 Installing the Micro Collector Spotter Site Requirements A su
94. spotter Accessory Kit Contents G1364 68715 Description Quantity Part Number Sticking clamp for corrugated waste tubing large 3 5065 9976 Sticking clamp for waste tubing small 3 5065 9976 ESD wrist strap 1 9300 1408 micro collector spotter Capillary Assembly 25 um ID 1 61364 87304 micro collector spotter Capillary Assembly 100 um ID 1 G1364 87306 ZDV Union stainless steel 1 5022 2184 Strip with 10 MALDI Spotting Tips 1 61364 81701 Plastic Spacers 20 not orderable Plug for waste container 1 G1364 26105 Reorder gives 5m t Reorder gives 10 clamps for corrugated waste tubing large and 10 clamps for waste tubing small Reorder gives 10 clamps for corrugated waste tubing large and 10 clamps for waste tubing small The Accessory kit G1330 68705 shown in Table 6 is shipped with the G1330B thermostat module if the thermostat module was ordered Table6 Optional Thermostat M odule Accessory Kit Contents G1330 68705 Description Quantity Part Number Waste tubing 1 2 m 1 5062 2463 Cable thermostat module to micro collector spotter 1 G1330 81600 Reorder gives 5m Agilent 1200 Series Micro Collector Spotter User M anual Installing the Micro Collector Spotter 1 Optimizing the Stack Configuration If your Agilent 1200 Series micro collector spotter is part of a system you can ensure optimum performance and minimum delay volume by installing the following configuration Figure 1 on page 18 sh
95. t will initialize the micro collector spotter Settings More The Fl key More opens a pull down menu Controlled Max Peak Duration 6 86 min 3Recovery on Track Detectors Agilent 1200 Series Micro Collector Spotter User M anual 71 3 72 Control M odule Screens of the Fraction Collector By selecting More Auxiliary on the pull down menu you can enter the micro collector spotter Fill Volume Per Vessel for the currently installed well plates or MALDI targets Leaving this value at the DEF value will completely fill the vessels according to their specified size Changes must be acknowledged with F6 Done By selecting More Flow you can specify the flow rate if no Agilent 1200 Series pump is configured in LC system The flow rate is required to calculate the needle movement in the liquid contact control mode and to calculate delay time between peak detection and collection of the fraction With an Agilent 1200 Series pump the flow rate will be automatically transferred to the micro collector spotter Changes must be acknowledged with F6 Done By selecting More Recovery on Track on the pull down menu you can activate the recovery mode of the micro fraction collector spotter With Recovery on Track selected the module will collect recovery fractions into the well plate if no peak is detected As soon as the next fraction is triggered the Agilent 1200 Series Micro Collector Spotter User M anu
96. tains e peek coated fused silica capillary 125 um ID 550mm length G1375 87318 Agilent 1200 Series Micro Collector Spotter User M anual Configuration and Operation of the Micro Collector Spotter 2 e T type connector 5042 8519 e Connector syringe to capillary 5042 8517 To setup the flow connections the outlet capillary from the UV detector the fraction collector capillary and the capillary from the matrix pump have to be connected to the T type Connector as displayed in Figure 14 on page 59 We recommend using e the 50 um ID fraction collector capillary G1364 87305 for overall flow rates LC flow and matrix flow lt 8 l min e the 100 um ID fraction collector capillary G1364 87306 for overall flow rates LC flow and matrix flow gt 8 l min The peek coated fused silica capillary 125 um ID 550mm length is used to deliver the matrix solution from the syringe to the T type connector where it is merged with the flow from the Agilent 1200 Series LC system The maximum spot size strongly depends on the type of the MALDI plate and can range between 0 5 and 5 ul The flow rate has to be adjusted to optimize the performance with spotting rate of 20 spots min maximum flow rate maximum spot size x 20 spots min Table5 Examples for maximum flow rates maximum spot size maximum spotting rate maximum flow rate LC flow matrix flow 0 5 ul 20 spots min 10 ul min lul 20 spots min 20 ul min 2 ul 20 spots min 40
97. tall substitute parts or make any unauthorized modification to the instrument Capacitors inside the instrument may still be charged even though the instrument has been disconnected from its source of supply Dangerous voltages capable of causing serious personal injury are present in this instrument Use extreme caution when handling testing and adjusting Agilent 1200 Series Micro Collector Spotter User Manual 145 A Safety Information Safety Symbols Table 21 shows safety symbols used on the instrument and in the manuals Table 21 Safety Symbols Symbol Description The apparatus is marked with this symbol when the user should refer to the AN instruction manual in order to prevent risk of harm to the operator and to protect the apparatus against damage Z Indicates dangerous voltages Indicates a protected conductor terminal l Eye damage may result from directly viewing the light produced by the Xenon flash ag lamp used in this product Always turn the xenon flash lamp off before removing it WARNING A warning alerts you to situations that could cause physical injury or damage to the equipment Do not proceed beyond a warning until you have fully understood and met the indicated conditions CAUTION A caution alerts you to situations that could cause a possible loss of data Do not proceed beyond a caution until you have fully understood and met the indicated conditions 146 Agilent 1200 Series Micro
98. tant temperature conditions in the module 102 Table8 To perform a position accuracy calibration for M ALDI targets Step Note 1 Switch to the Diagnosis View of the Agilent ChemStation Software 2 Select Tests from the Diagnosis menu Agilent 1200 Series Micro Collector Spotter User M anual Troubleshooting and Test Functions 4 Table8 To perform a position accuracy calibration for M ALDI targets continued Step Note 3 Inthe Test Selection dialog box select Micro AFC G1364D from the drop down list 4 Select position accuracy calibration M ALDI and click on Start 5 Start the position accuracy calibration and Click on Explain to get detailed instructions follow the instructions on the screen and information 6 Align the xy position of the capillary tip tothe See Figure 20 on page 104 center of each corner position of the MALDI for Agilent Technologies AP M ALDI Target or to each pinhole on the MALDI targets use calibration plate 5023 0214 calibration plates with the corresponding for Micromass M ALDI targets use positioning buttons calibration plate 5023 0215 for Bruker M ALDI Targets use calibration plate 5023 0208 for Applied Biosystems M ALDI targets use calibration plates 5023 0209 or 5023 0213 If a calibration plate is not available use the M ALDI Target and align the capillary to the center of each corner position 7 Align the height z position of the capillary tip The capillary ti
99. ted conditions are fully understood and met A WARNING notice denotes a hazard It calls attention to an operating procedure practice or the like that if not correctly per formed or adhered to could result in personal injury or death Do not proceed beyond a WARNING notice until the indicated condi tions are fully understood and met Agilent 1200 Series Micro Collector Spotter User M anual In This Guide This manual contains user information about the Agilent 1200 Series micro collector spotter G1364D It is intended for users that will operate the Agilent 1200 Series micro collector spotter either for micro fraction collection or for MALDI Spotting For information about installation advanced troubleshooting repair and a complete list of internal parts please refer to the Agilent 1200 Series Fraction Collector Service Manual G1364 90110 1 Installing the Micro Collector Spotter This chapter provides information about the installation of the Agilent 1200 Series micro collector spotter 2 Configuration and Operation of the Micro Collector Spotter This chapter describes the configuration method setup and operation of the micro collector spotter 3 Control Module Screens of the Fraction Collector The reference to the screens of the Agilent 1200 Series control module is provided in the chapter 4 Troubleshooting and Test Functions This chapter describes the modules built in troubleshooting test and maint
100. th 39 Performance 140 performance specifications 139 analytical scale 140 physical specifications 12 fraction collector 12 thermostat 13 plates 130 position accuracy calibration 39 97 power consideration 10 power cords 11 power requirements 10 power supply indicator 90 preview 51 R recovery 49 recovery location 83 recovery on the track 49 removing tray compartment divider 25 repairing the fraction collector 107 repairs cleaning the fraction collector 108 main procedures overview 109 simple repairs 108 report 58 reserved locations 46 83 reset 37 S safety information on lithium batteries 147 sensor pad 34 shipping 32 simple repairs 108 109 site requirements 10 slope only 48 specifications 12 139 spotting adapter 40 Agilent 1200 Series Micro Collector Spotter User M anual Index spotting rate 49 start location 54 status indicators 90 step commands 105 storage 12 T temperature 12 tests 84 thermostat electrical connections 26 threshold upper 48 tick marks 58 time based 49 timetable 47 48 transport 32 transport unit assembly 32 135 transport unit self alignment 95 transporting the fraction collector 32 tray combinations 28 tray compartment divider 25 trays 43 81 140 numbering of vessel positions 28 trigger modes 140 troubleshooting and test functions 89 U unpacking the fraction collector 14 upper threshold 48 Ww
101. the capillary is bent or blocked by particles 38 Agilent 1200 Series Micro Collector Spotter User M anual Configuration and Operation of the Micro Collector Spotter 2 Adjusting the peakwidth of your detector When using capillaries in these flow ranges for peak based fraction collection a Peakwidth of 0 1 min Responsetime lt 2s or less should be selected to ensure a fast signal processing Otherwise the compound might already be flushed through the micro collector spotter when the peak is detected If a smaller peakwidth is set the flow rate ranges can be increased Capillary Alignment for micro fraction collection During the initialization process the vertical and the horizontal position of the capillary tip is determined The micro collector spotter facilitates the capillary check by pressing the capillary onto the sensor pad Then the relative capillary length as well as the degree to which the needle is bent is determined 384 well plates require a higher position accuracy than 96 well plates Consequently a slightly bent capillary might be tolerated for 96 well plates whereas 384 well plates require the installation of a new capillary or a position accuracy calibration as described in Position Accuracy Calibration for 384 Well Plates on page 100 After the capillary check procedure a message is displayed if any further capillary alignment is required If solvents with high concentrations of matrix buffer or salt are use
102. the tray compartment After a leak in the sampler make sure the leak plane is cleaned and dry 144 Agilent 1200 Series Micro Collector Spotter User M anual Safety Information A Operation Before applying power comply with the installation section Additionally the following must be observed Do not remove instrument covers when operating Before the instrument is switched on all protective earth terminals extension cords auto transformers and devices connected to it must be connected to a protective earth via a ground socket Any interruption of the protective earth grounding will cause a potential shock hazard that could result in serious personal injury Whenever it is likely that the protection has been impaired the instrument must be made inoperative and be secured against any intended operation Make sure that only fuses with the required rated current and of the specified type normal blow time delay and so on are used for replacement The use of repaired fuses and the short circuiting of fuseholders must be avoided WARNING Any adjustment maintenance and repair of the opened instrument under voltage is forbidden WARNING Disconnect the instrument from the line and unplug the power cord before maintenance Do not operate the instrument in the presence of flammable gases or fumes Operation of any electrical instrument in such an environment constitutes a definite safety hazard Do not ins
103. tion 1 Capillary Up 3 Use the F1 key Align Trans to perform an automatic alignment of the micro collector spotter transport unit WARNING An empty 4 well plate tray G1364 84511 or G1364 84521 and the well plate adapter must be installed when performing the alignment The well plates must be removed from the micro collector spotter tray before performing the transport unit alignment If the well plates are installed during the alignment procedure the transport unit might be damaged The alignment process may take several minutes Changes must be acknowledged with F6 Done To exchange the capillary the well plate adapter or the capillary carrier assembly the arm can be moved to the F2 Change position where the user has access to these parts The tray has to be removed Before the arm can move to change position please remove the currently inserted tray Press to continue Align Transp Part Info Records Use the Fl key EMF to set EMF limits Changes must be acknowledged with F6 Done Agilent 1200 Series Micro Collector Spotter User M anual 85 3 Control Module Screens of the Fraction Collector Records Module Product Serial Version On Time G1323B8 CN23605001 B 04 02 AF Collector G1364D DE43640110 A 06 01 012 Od 00 14h If a set limit has been exceeded a message box will pop up If you press Reset the limits will be removed Ignore will continue to ke
104. tion collection working with up and down slope publication number 5988 7895EN The option to specify an Upper Threshold becomes important if the absorbance exceeds the linear range of the UV Vis detector At high absorbance values the light intensity on the detector is extremely low and consequently detector noise will superimpose the detector signal In this case the detector noise might lead to wrong trigger commands for the micro collector spotter As soon as the detector signal exceeds the Upper Threshold settings for slopes will be ignored until the signal drops again below the Upper Threshold When using more than one peak detector fraction collection can be triggered either when all selected peak detectors detect a peak or when atleast one selected peak detector detects a peak based on the settings in the detector table The Timetable can be used to program changes in the Fraction Trigger Mode during the analysis by entering a time in the Time field and appropriate values in the fields of the timetable The trigger modes can be specified as Off Agilent 1200 Series Micro Collector Spotter User M anual Configuration and Operation of the Micro Collector Spotter 2 Peak based and Time based Time ranges can be defined for the different trigger modes In the column headed Time If the Off mode is selected no fractions are collected Whenever the Peak based mode is specified fractions will be collected based on the peak detection p
105. to perform a Transport Unit Self Alignment Steps Comments 1 Ifthe transport unit has been exchanged or if it See Service Manual G1364 90110 for details is strongly misaligned set the 8 bit configuration switch to the Forced Cold Start Configuration 2 Install the 4 well plate tray G1364 84511 or IM PORTANT Remove all plates G1364 84521 3 Ensure that the wellplate adapter is correctly assembled Agilent 1200 Series Micro Collector Spotter User M anual 95 4 Troubleshooting and Test Functions How to perform a Transport Unit Self Alignment Steps Comments 4 Select the Maintenance menu in the Diagnosis view of the Agilent ChemStation 5 Inthe menu choose Fraction Collector gt The Transport Alignment Procedure takes Transport Alignment to startthe automated approximately 10 15 minutes procedure 6 Setthe 8 bit configuration switch to the default See Service Manual G1364 90110 for details setting 96 Agilent 1200 Series Micro Collector Spotter User M anual Troubleshooting and Test Functions 4 Arm Length Calibration The Arm length calibration is only be required for 384 well plates This procedure calibrates for the arm length of the capillary carrier assembly The arm length calibration is required e after disassembling the transport unit capillary carrier assembly e if problems with the positioning of the capillary tip occur in particular on plate 4 back right We recommend to switch
106. tted before the fraction collector Auxiliary In the Auxiliary section the Maximum fill volume per location can be specified If as configured is selected the pre configured volume see Instrument gt Pre configured Wellplate Types is used This ensures that the location well vial or tube cannot be overfilled during fraction collection This volume can be further reduced by defining a customized volume By default all MALDI plates are configured with an infinite fill volume The Maximum fill volume can be used to determine a maximum spot size if the peak based trigger mode is used Agilent 1200 Series Micro Collector Spotter User M anual Configuration and Operation of the Micro Collector Spotter 2 Setting up a Micro Collector Spotter Method Additional check boxes in this section provide the opportunity to Store the temperature signal and the UIB signal Fraction Preview To determine the appropriate fraction collection parameters the Agilent ChemStation provides a valuable tool that becomes accessible by selecting the button labelled Fraction Preview Tool Figure 7 in the Peak Detectors section Figure 7 Fraction Preview dialog box The Fraction Preview screen allows testing the fraction collection parameters against a sample chromatogram It can also be used to optimize the fraction collection parameters interactively With the help of this tool values for up and down slope as well as for upper and lower threshold can e
107. ul min 5 wl 20 spots min 100 ul min Contact Control Adjustment If the matrix is added online the flow that is delivered by the Agilent 1200 Series capillary or nanoflow pump is different from the flow at the capillary tip in the MALDI spotter To account for this additional flow the Contact Control Adjustment has to be specified as described on page 50 Agilent 1200 Series Micro Collector Spotter User M anual 61 2 Configuration and Operation of the Micro Collector Spotter Online M atrix Delivery CAUTION If solvents with high concentrations of matrix buffer or salt are used the capillary has to be flushed thoroughly with salt free water after analysis Such a procedure prevents the capillary and the waste port from clogging 62 Agilent 1200 Series Micro Collector Spotter User M anual Configuration and Operation of the Micro Collector Spotter 2 Check out procedures The purpose of the check out injection is the verification of the correct installation and configuration of the Agilent 1200 Series micro collection spotting system In addition the checkout procedure can used for troubleshooting if the Agilent 1200 Series micro collector spotter doesn t operate as expected The checkout procedure depends on the application and is different for micro fraction collection into well plates and for MALDI spotting Check out procedure for micro fraction collection The check out procedure is based on the procedure for the
108. view F4 from the Analysis Screen you can specify the sequence s End Actions F1 load sequences from PC Card F2 if present or proceed to a method table F3 F6 Delete and F7 Insert allow you to edit lines in the sequence table Line From To Inj Vol Method Wait Calibration end of sequence ee ORR oe ne iis 0 00 Ready On end of Cl Load Method ACTUAL z Sequence Tum off Timeout ejaa min Turn off complete HPLC system Parameters that can be entered into a line in the sequence table ace CMe lt lt RUE ace 0 O00RFIE Ready Viallil__ to __1 njC T Vol DEFH ul Method ACTUAL Wait after loading min 78 Agilent 1200 Series M icro Collector Spotter User M anual Fractions Control Module Screens of the Fraction Collector From the Analysis View press the F5 key Views and select Fractions Flow rl sce A 1 Analysis 2 000 2 Samples BCO This is an example if an Agilent 1200 Series micro collector spotter is configured stand alone Here you can define a Start position for a fraction select a collection Scheme choose a Trigger Mode etc i Fraction L Scheme 63 cS O lil oll Agilent 1200 Series Micro Collector Spotter User M anual 3 79 3 Control Module Screens of the Fraction Collector Screens available from the
109. ying in contact with the surface of the collected liquid For low flow rates lt 1 ul min a small Distance lt 0 2 mm should be chosen This value should be increased to 0 3 0 7 mm in particular for MALDI spotting at higher flow rates The capillary movement in the Contact Control mode depends on the flow rate that is delivered by the Agilent 1200 Series pump If your LC system contains more than one pump the flow rate of the first pump in your system determines the needle movement In this case verify in your system configuration Instrument gt Configure 1200 access that this pump is the first one in the Configured M odules list In addition this pump should also be selected as main pump Instrument gt Change main pump Well Plates In the Well Plates section the type of well plates or MALDI targets used in a well plate tray can be configured The well plate type used can be chosen from the Plate Type dropdown menu More detailed information about all preconfigured well plates and MALDI targets can be found in the Instrument menu A list of recommended Agilent well plates can be found in Table 12 on page 130 and a list of supported MALDI targets is available in Table 16 on page 133 To configure other well plates or MALDI targets choose Configure Well Plates from the Instrument menu Note the orientation of the well plate or the M ALDI plate The starting position Al is always at the back left corner of the plate Only one type of
110. ys F7 F8 Remove Add to move the highlighted module Changes must be activated with Done F6 Here e g the Agilent 1200 Series pump the injector and the detector parameters are shown on the display as well The number of parameters for each module is reduced as additional modules are added to the display A maximum of 4 modules can be shown simultaneously If more modules are connected to the system you have to choose 4 of them in Setup view Flow ni A 0 000 Samp Ref With the F1 key Settings you open a pull down menu where you can select the micro collector spotter module Within Settings you can change the type of fraction collection Off Timetable or Peak Controlled There are different sets of parameters available through the F1 3 and F5 navigation keys for setting path operation of the micro collector spotter F7 Default resets the micro collector spotter to default values Changes must be acknowledged with F6 Done Agilent 1200 Series Micro Collector Spotter User M anual Control Module Screens of the Fraction Collector 3 Peak Controlled lt fiex Peak Duration Settings screen with thermostat connected On Off F8 allows you to switch the thermostat on or off Settin aS ine I Loc REE Inj Haus 60 OOM Ready Turn Temp On Off Use the m key for the context sensitive menu The Status command pulls up a module specific screen displaying details of configured trays Rese

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