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PowerWaveX Operator`s Manual - Bio-Tek

1. EPORT UTIL SELECT UTILITY OPTION TESTS SETUP OUTPUT RINT REPORT ESULT MAP ASSAY LIST ECT ASSAY NUMBER 01 E RAW_OD E RAW_OD Figure 3 2 Options available from the Main Menu Operation DEFINE The Main Menu option DEF INE allows you to define the data reduction parameters for a new assay or to modify a previously defined assay stored in memory EPORT UTIL ECT ASSAY NUMBER 01 E RAW_OD Press Options to scroll through the assay list Press ENTER to select an assay See Editing the Assay Name on page 3 7 for instructions on modifying the current name Press ENTER to continue FORMULA CURVE Figure 3 3 Options available under DEFINE e METHOD Define the method of reading such as Endpoint Kinetic or Scan and Single or Dual Wavelength Specify a delay before read or incubation settings e MAP Specify the plate layout using blanks controls standards and or samples Choose to map the plate manually or let the software map it automatically e FORMULA Define cutoff transformation validation and or general formulas Create variables to be used within formulas e CURVE Specify a curve fit type and x y axis types lin log Specify the method by which standard outliers can be edited Enable or disable the extrapolation feature PowerWaveX Operator s Manual 3 5 SELECTING AN ASSAY
2. Assay Validation formula s establish a set of criteria used to determine whether or not an assay can be considered valid If the criteria are not met results are considered suspect and the message RESULTS INVALID Assay validation failed appears on results reports e Up to four assay validation formulas can be defined e Define the plate map via DEFINE MAP before creating assay validation formulas Examples If an assay protocol states that in order for the assay to be valid e The mean of the negative controls must be less than 0 100 The formula is NC x lt 0 100 e The mean of the positive controls must be greater than the mean of the negative controls The formula is PC xX gt NC x 3 34 Operation TRANSFORMATION FORMULAS Transformation formulas can be used to transform raw or blanked absorbance data in preparation for further data reduction including curve fit analysis See Formula Type on page 3 30 for instructions on selecting an assay and accessing the Transformation Formula definition screen e Ifa blanking method is selected in the assay transformation formulas are applied to the blanked absorbance values otherwise they are applied to the raw data Turn to page 3 29 to review the results calculation structure e One transformation formula may be defined per assay e A transformation formula can be simple ex OD 100 to multiply all wells on the plate by 100 or more complex
3. Specifying the Wavelength mode Single or Dual e If SINGLE is chosen the reader measures the optical density of each well with a single wavelength e If DUAL is chosen each well is read twice at two different wavelengths The microplate is not removed from the reading chamber between the two measurements The final reported optical density is the difference between the two readings Note Dual wavelength readings can significantly reduce optical interference caused by scratches or fingerprints that absorb light equally at both wavelengths Choose a REF wavelength that is far from the MEAS wavelength in the spectrum for best results MEASUREMENT WAVELENGTH S After specifying Single or Dual Wavelength the MEAS or MEAS REF selection screen appears for wavelength selection Pre rra ree COBRA O EA ERA AE Specifying the wavelength s to be used during plate reading Single wavelength selection is shown on top Dual wavelength selection on bottom PowerWaveX Operator s Manual 3 11 e If reading at a single wavelength a MEASurement wavelength must be selected If reading at dual wavelengths a REFerence wavelength must also be selected e Choose a wavelength value from those presented Select MORE to see additional wavelengths e Note Ifa desired wavelength is not listed the reader s internal wavelength table must be changed Press the Main Menu button to save the current assay settings and ret
4. 3 22 Operation Blank Location The Blank Location screen is presented if blank wells are defined and manual map generation is selected e ee ee BME GTA II eel Defining the Location of the blank wells e Use the NUMERIC and ALPHA keys to enter the location of the first Blank well e Press ENTER to define subsequent Blank well location s Number of Standards For assays requiring standards begin by entering the number of standard groups Entering the number of standard groups e Use the NUMERIC keys to enter the number of standard groups gt Enter 00 to indicate no standards e The valid entry range depends on the selected curve fit method The maximum number of standards for all curve fit methods is 12 The minimum numbers of standards are gt 4 for 2 P 4 P cubic cubic spline gt 3 for quadratic gt 2 for linear point to point e Note Ifthe NUMBER OF STANDARDS setting is modified the number of standard replicates see next page automatically defaults to 01 PowerWaveX Operator s Manual 3 23 Number of Standard Replicates The Standard Replicates entry screen is presented if the number of standard groups is greater than 0 Entering the number of standard replicates e Use the NUMERIC keys to enter the number of standard replicates The valid entry range is from to 8 replicates gt The software will automatically verify that the number of replicates multiplied by the number of stand
5. 3 6 From the Main Menu Assay Number screen press the soft key beneath the DEF INE menu option to access the Select rele Bele Vee se Mee eae al parra ee Assay Selection screen showing the current assay name and number e Use the NUMERIC keys to enter the number of any predefined assay stored in the reader s memory or the OPTIONS key to advance one assay at a time The cursor is positioned at the first editable field and advances automatically The numeric range depends on the number of assays programmed in the reader s memory The reader typically e Press EN has 55 open assays available TER to advance to the Edit Assay Name screen You may change the default assay number to a more descriptive one see Editing the Assay Name on the following page gt gt gt gt CLEAR Clears the reader s display MAIN MENU Returns the display to the Main Menu screen PREVIOUS SCREEN Returns the display to the previous screen ENTER Saves the current settings and advances to the next screen Note Within every screen these keys will continue to function as described above Operation EDITING THE ASSAY NAME Use the Edit Name screen to edit the name currently assigned to the assay The assay name can contain up to 16 alphanumeric characters ee ea Pre PO cil ey Pe ee el ET IIe eT ee Edit Assay Name screen e The cursor is positioned at the first edi
6. First Page of Column Report Showing Plate Data for Blanks and Standards the Interpretation of Results and Comments ccsceessesseeeseeeseeeeceteeeseneeneenseenaes D 4 Second Page of Column Report Showing Plate Data for Samples ceceeeee D 5 Column Report Without Samples ccccccssccsseeecseeeeeeeeeesceeseeeeeseeneeeeenseenseesaeeaees D 6 Panel Report aia D 7 Assay Detail Reporta e y bb D 8 ASSAY ii A Ba aOR eee ein tea ee ee D 9 PowerWaveX Operator s Manual xxi List of Tables xxii Table 2 1 Table 2 2 Table 4 1 Table 4 2 Table 4 3 Table 4 4 Table 4 5 Table B 1 Table B 2 Parallel Connector Pinouts ccceseccsesecseeeeeseceeeceeeeceaeeeeesecsessecneeeeeeaeeeeeaeeas 2 7 Serial Pinout Description nnie 2 8 Recommended Test Schedule ceeccessecssecseeeecneveeceseeeeesecaeeseceeveceaeeeeeaeeaeaes 4 2 Typical Enzyme Substrate Combinations and Stopping Solutions 4 14 Stock Solution Formulation for Liquid Test Nos 1 and 2 0 0 ce cceseesesteereees 4 15 Test Tube Dilutionsic iia id ii 4 19 Phosphate Buffered Saline 10X Concentrate Solution oooonoonnnnncionmmmmmmm 4 22 Serial Cable Pin Out Description cccceccceccesseesseeeeeeeeeeeceeeeeeeesecereneeeeeseneensees B 3 ASCII Control Characters Used in Computer Control Protocol B 6 Preface M Chapter 1 Introduction This chapter introduces the PowerWaveX a
7. oooooccccncnnonococccnnccoconononccnnnnnnnnnnoncnnncnnnnnnnnnnnncnnnnnannnnns 2 8 Setting Up Communications Parameters ooooccnnnnncccnonoccncnonononnnancnnnnn cnn nnnnn cnn nan nnnnnnns 2 9 COMPONEN Sisipan cri tae tree sn 2 11 Laia l lt 2 11 GOGlING Farr uri tddi 2 11 Adjusting the Line Voltage Input Range ooooocccccccncocococononocaconononononcnnnnnononocnncnnnnannnncnnno 2 12 Adjusting the Fuse Configuration or Fuse Replacement ccccesseeeseeeeeeesneeeeeeneees 2 14 Other Utility Options ccoo dev ii we 2 15 Setting the Date and Time cccccccecceeceeeeeeeceeeeceeeeesecneaeeeeeeeseseeaeeeeeeeeeeeenaees 2 16 Specifying Data Output and Reporting Options oooococccnnnnnicocccccccccccnnancncconcnanann nos 2 17 Data QU PUtacia SA end Ne el ee ee eee eal 2 17 Compatible PriNtelS iio 2 17 Report POS te ani one tani eer a hates 2 18 Selecting Read Options ssns SR dee Sten bene ss eos 2 19 Chapter 3 Operation ices iccciccccscasvecucetvecesceceseucesvisevescasstcsdeschsedensesersedeuce covers 3 1 OVO IVIOW iiini i cen ck e E Ea aaa a E EA iaai 3 1 Front Paneleciienini ane a a ea a E a bead a aa ES 3 2 Selecting Menu FUNCTIONS ieissar niei eiii 3 2 Scrolling Through Options ooooccconoccccnonoccnonononanononcncnnn nn nc nono cnn nano nnnnnnn nn nr nnn cnn nan nnnnnnns 3 2 NES 3 3 Moving Backward Through the Menus cooccccnnoccccnonoccnonononcnanoncno nano cnn n non nc nnnnnnnnnnnoss 3 3 SAVING ASSAYS O et
8. 0 500 CTL CTL2 x gt CTLI x CTL3 CTL3 x gt CTL2 x CTL4 CTL4 x gt CTL3 x Sa E AAA AR Well 1D 81kd0D Calc0D Call Pred Conc SLT Std Dev CVA Notes A A ee ee A01 BLK 0 030 0 030 B01 BLK 030 030 AVE 00 00 0 043 error cot CTLi 0 311 0 311 INV D01 CTLI 0 333 0 333 INV AVE 0 322 0 322 0 016 4 926 EQ CTL2 0 517 0 517 VAL FOL CTE 0 554 0 554 VAL AVE 0 535 0 535 0 026 4 782 601 CTL3 0 814 0 814 VAL HOt CTLI 0 857 0 857 VAL AVE 0 835 0 835 0 030 3 636 A02 CTL4 1 118 1 118 VAL B02 CTL4 1 092 1 092 VAL AVE 1 105 1 105 0 018 1 667 End of Report Figure D 5 Column Report without Samples D 6 Sample Reports Assay PANEL COMMENTS Specimen Assay SMP0001 ASSAY 01 ASSAY 02 ASSAY 03 SMP0002 ASSAY 01 ASSAY 02 ASSAY 03 SMP0003 ASSAY 01 ASSAY 02 ASSAY 03 SMP0004 ASSAY 01 ASSAY 02 ASSAY 03 SMP0005 ASSAY 01 ASSAY 02 Page 1 Bio Tek Instruments Well B1Kd0D 601 HO1 AVE 605 H05 AVE G09 H09 AVE A02 B02 AVE A06 B06 AVE At0 B10 AVE C02 D02 AVE C06 D06 AVE C10 D10 AVE E02 F02 AVE E06 F06 AVE E10 F10 AVE G02 H02 AVE G06 HOG AVE PowerWaveX Operator s Manual 0 729 0 709 0 719 662 695 679 402 361 1 382 211 1 205 1 238 0 818 0 850 0 834 1 686 1 653 1 669 0 719 0 744 0 731 296 357 326 1 173 190 181 069 090 080 2 606 2 626 2 616 1 205 1 121 1 163 1 367 1 076 1 222 2 465 2 538 2 502 Date 06
9. 100 This divides all the wells by STD1 presumably the 0 standard and multiplies the results by 100 To do this gt At the Scope Variable selection screen choose OD and press ENTER gt Enter simply STD1 as the TVAR formula Press ENTER gt The formula selection screen is displayed Choose TRANS gt Enter the formula OD TVAR 100 using the Math Other Map and Function keys TVAR is available as a MAP option PowerWaveX Operator s Manual 3 37 CUTOFF FORMULAS A cutoff formula calculates a cutoff value that is used for classifying samples See Formula Type on page 3 30 for instructions on selecting an assay and accessing the Cutoff formula definition screen During data reduction results are evaluated against the cutoff value with an optional greyzone and each well is assigned a call POS positive NEG negative or EQUIV equivocal Greyzone One cutoff formula may be defined per assay If Transformation Formulas are defined cutoffs are based on the transformed results Turn to page 3 29 to review the results calculation structure A cutoff formula can consist of a simple numeric value 1 500 a well identifier PC or PC x to represent the mean or a formula combining the two NC x 0 050 A greyzone around the cutoff value can be defined to indicate questionable results Do not use the lt or gt mathematical symbols in a cutoff formula Tip Choose to print a Column Report to se
10. 200 999 ASCII Computer Control host 70 84 85 162 163 164 165 166 167 168 169 170 valid limits response remaining wavelengths 000 999 ASCII don t care shake time seconds 0 999 shake speed 0 slow 1 med 2 fast 3 variable total kinetic read time minutes 9999 delay before first read 0 no 1 yes delay seconds 0 999 status string 5 Up to six wavelengths may be specified for a single plate read definition The assay definition table must be downloaded before other plate specific commands are sent such as Set Plate Geometry and Set Strip Range Ifa wavelength is not to be used it must be replaced by 000 GET WAVELENGTH TABLE W This command uploads the wavelength setup table from the reader host Ww valid limits response no arguments lt ACK NAK gt Reader response protocol ASCII format for each wavelength configured six wavelength comma terminator status string 5 PowerWaveX Operator s Manual 000 999 gt HALT X This command causes any read or spectral scan in progress to be halted host valid limits response x no arguments none see below no status string The following events occur when this command is invoked 1 The scan or read process is halted and all axes are returned to their home positions the plate is be mo
11. S shaped plot that eventually becomes asymptotic to the upper and lower standard values The logistic equation is algebraically transformed to a simpler form in which experimentally determined values are used for the responses at concentrations of zero and infinity C SPLINE Cubic Spline A piecewise polynomial approximation consisting of joining a set of data points by a series of straight lines which is then smoothed by using a cubic fit 4 P 4 Parameter Logistic A curve fitted to the standard values which is characterized by a skewed sigmoidal S shaped plot that eventually becomes asymptotic to the upper and lower standard values The 4 parameters are Left asymptote Right asymptote Slope and Value at the Inflection point This fit is most recommended for immunoassay data and is more exact than Logit Log PT PT Point to Point A plot that connects each standard point with a line with no averaging of the values to smooth the curve at each standard EDIT STANDARD OUTLIERS After the standard curve has been generated one or more standards can be excluded from the recalculation of the curve Choosing whether or not to enable standard outlier editing for this assay 3 42 Select NONE to suppress the Edit Standard Outliers capability for this assay Choose MANUAL to enable the capability gt IfAVERAGE STANDARDS is set to NO the individual standard replicates are available for e
12. Use the formula keys Math Other Map and Function to define the Transformation Variable TVAR Once the variable has been defined it can be used in a transformation formula The TVAR will be available as a MAP option when building the transformation formula Example The assay plate map has 2 blanks 1 control well in duplicate CTL1 1 negative control well in triplicate NC and 5 standards in duplicate STD1 STDS in varying concentrations The assay data reduction states gt Subtract the mean of CTL1 from the mean of the NC Subtract the difference from all OD s on the plate gt Divide the result of the above by the mean of the NC less the mean of CTL1 then multiply by 100 On paper the formula reads OD NC x CTL1 x NC x CTL1 x 100 On the reader the formula NC x CTL1 x will be programmed as the TVAR since the transformation will apply to all standards controls and samples on the plate To do this gt At the Scope Variable selection screen choose OD and press ENTER gt Enter the formula NC x CTL1 x using the Math Other Map and Function keys Press ENTER 3 36 Operation gt The formula selection screen is displayed Choose TRANS gt Enter the formula OD TVAR TVAR 100 using the Math Other Map and Function keys TVAR is available as a MAP option Example In the case of competitive reactions converting absorbance data to percent B Bo can be OD STD1
13. 080 1 816 2 475 1 316 0 768 1 274 0 954 0 737 1 049 Well STD2 STDE SMP3 SMP7 SMP11 SMP15 SMP19 SHP23 SMP27 SHP31 SMP35 SHP39 RSLT 21 416 57 558 47 165 39 488 69 631 eee 46 984 29 315 45 629 35 488 28 232 38 524 G CALL NEG POS POS POS NEG NEG POS NEG NEG POS CalcOD 0 818 1 873 0 777 1 275 1 519 2 347 0 822 0 774 1 241 0 755 0 790 1 286 Well 103 S107 SMP4 SMPB SMP 12 SMP 16 SHP20 SHP24 SMP28 SMP32 SMP36 SMP40 RSLT 31 012 ep 29 619 45 671 53 985 k ka 31 151 29 509 44 582 28 869 30 082 46 007 _ o _ _ ji _ ____ _ _ _ _ CALL NEG POS POS POS NEG NEG EQUIV NEG NEG POS CalcoD 0 853 1 914 0 758 1 248 1 495 2 394 0 767 0 810 1 183 0 802 0 773 1 303 Well STD3 STD SMP4 SHPB SMP12 SMP16 SMP20 SMP24 SMP28 SMP32 SMP36 SMP40 RSLT 32 187 eee 28 969 44 808 53 100 eek 29 288 30 745 42 727 30 469 29 498 46 578 _ _ HHHHHAAA A A AAA A _ A cas Figure D 1 Matrix Report showing well IDs calculated OD values and concentrations calls on samples and comments Sample Reports Bio Tek Instruments Assay ASSAY1 Date 07 01 99 Lot Time 10 36 10AM Operator Wavelength 405 Temp 30 C Plate 1D 123 COMMENTS Some concentrations may lie outside curve asymptotes lt gt around RSLT indicate extrapolated concentration Standard Curve Quadratic Fit 1 894 m o Z b ww FO woo eS t 0 307 10 000 S000 ae
14. 2 500 Abs 3 0 000 to 2 000 Abs 2 2 000 to 2 500 Abs 3 0 000 to 2 000 Abs 2 2 000 to 2 500 Abs 3 0 010 Abs 0 010 Abs 0 010 Abs 0 010 Abs Spectral Scanning or Kinetics with short intervals When read intervals from 32 seconds to 80 seconds are selected or spectral scanning is performed the specifications are as follows Absorbance Measurement Range 0 000 to 4 000 Abs Accuracy Linearity Repeatability 0 000 to 2 000 Abs 1 2 000 to 2 500 Abs 3 0 000 to 2 000 Abs 4 2 000 to 2 500 Abs 4 E 2 E 3 0 000 to 2 000 Abs 2 3 2 000 to 2 500 Abs 0 010 Abs 0 010 Abs 0 010 Abs 0 010 Abs Introduction e Hardware and Environmental Specifications Display Light Source Dimensions Weight Environment Humidity Power Consumption Incubation option Internal bar code reader option Robotic interface option PowerWaveX Operator s Manual 2 X 24 Character LCD Xenon flash light source 10 W max average power Life 1 billion flashes 16 0 deep X 15 5 wide X 9 75 high 40 1 cc deep X 39 3 cc wide X 24 8 cc high 40 lb maximum 18 2 kg Operational temperature 18 40 C 10 to 85 non condensing 100 VA Four Voltage Ranges accommodated by voltage selection switch Range 1 100 VAC 90 110 VAC 50 60 Hz Range2 115VAC 103 127 VAC 50 60 Hz Range3 230VAC 207 253 VAC 50
15. 2 of 2 Page 4 2 Recommended Test Schedule Correction The schedule shown in Table 4 7 defines the factory recommended intervals for performance testing for a microplate reader used for one shift seven days a week Note The risk factors associated with your tests may require that the Operational and Performance Qualification procedures be performed more or less frequently than shown below Table 4 1 Recommended Test Schedule Operational Performance Performance Installation Qualification Qualification Qualification Oualification Initially amp Annually Monthly Quarterly Liquid Test 1 p 4 17 or Liquid Test 2 p 4 19 Liquid Test 3 p 4 22 Robotic Lube p 4 25 Every six months or after 10 000 cycles Ifyou have an Absorbance Test Plate run Liquid Test 1 If you do not have an Absorbance Test Plate run Liquid Test 2 Liquid Test 3 is optional it is provided for sites requiring verification at wavelengths lower than those attainable with the Absorbance Test Plate Liquid Tests should be performed as part of the Operational Qualification during initial installation and annually as shown in this revised chart BIO TEK BIO TEK INSTRUMENTS INC P O BOX 998 HIGHLAND PARK WINOOSKI VERMONT 05404 USA COPYRIGHT 2005 TEL 888 451 5171 FAX 802 655 7941 Outside the USA 802 655 4740 E mail customercare biotek com www biotek com Creating Solutions for the Future of Science PowerW
16. 3 3 Starting or Stopping a Read cooocccococcccconoccnonononcnnnonononano nono nono cnn nan nn nn nana n cnn rnnn rra nan nnnnnnns 3 3 SHAUN ia 3 3 Main Melia A A de a 3 4 DEFINE cocina aa decade 3 5 Selecting AMAS ot ir RA td 3 6 Editing the Assay Name ccccccceesesceceeeeeeeeeeeeeeeeeesecaaeaeeeeeeesecacaeeeeeeeseeenaeeeeess 3 7 xvi Preface Define METHOD a aa 3 8 RO PO ii A A AE 3 8 Spectral SCANNINO cita rd 3 9 Delay First REA etico iii 3 10 Incubation Temperature Reader Dependent occccnnoccccnonoccnononcncnononcncnana nano nancninns 3 10 WaVvelengih S isiicaroia iba fail 3 11 Measurement Wavelength S ooococinccccccnnnccconononcconcnccnnononcnnnnnnnnnnnn nc nn cnn nnnnnnnnnnnn 3 11 Kin etic Read ParaMeters ooococoiicc tinc cerda 3 12 Kinetic Intervalo a 3 12 Total Number of Kinetic Reads oooocccnnocccccococcnononcnononancnn nono nonn acc nc nana nnnrnnnnnnnns 3 13 Kinetic Duration 0 00 ceeeceeeeeneeeeeeneeecneeeeeesaeeeeeeeaeeesneeeeeenaeeeeseneeeeesnaeeeeneaas 3 13 Plate ShakiNGi ecioirioa cs A E 3 13 Shake TIM cia aia 3 14 Shake Speed anani dia 3 14 Kinetic AmalySis iio arcos 3 15 Kinetic Pot A dead 3 15 Onset OD iii A rs 3 16 Well SCannind ita ai nah Uae head 3 16 Define MAP caidos sez Se eta reo roto de cnt dde eta are La o Std SP asta e ala abd 3 17 Map Generation idos 3 18 Mapping Direction viviusnomaa aii 3 18 Replication Direction acia iii ia 3 19 Start Mapping at Well Loca
17. 320 184 644 816 730 519 495 507 509 672 590 148 Call POS POS POS NEG NEG NEG POS POS POS NEG NEG NEG EQUIV EQUIV EQUIV NEG NEG NEG EQUIV EQUIV EQUIV POS POS POS NEG NEG NEG EQUIV POS EQUIV POS POS POS POS POS POS NEG NEG NEG EQUIV Pred Conc RSLT 47 526 46 914 47 219 28 703 30 670 29 694 47 582 47 165 47 373 29 619 28 969 29 295 38 776 41 505 40 142 31 254 31 070 31 162 39 966 39 488 39 727 45 671 44 808 45 239 29 703 30 631 30 179 38 467 47 118 42 760 59 017 69 631 63 294 33 985 33 100 53 540 19 394 25 099 22 774 41 629 Std Dev 0 013 0 040 0 009 0 013 0 061 0 004 0 011 0 019 0 020 0 193 0 122 cv 1 001 5 183 0 681 1 727 5 566 0 471 0 988 1 511 2 473 16 275 7 041 1 132 19 626 Figure D 4 Second page of Column Report showing plate data for samples Notes Bio Tek Instruments Assay ASSAY3 Date 07 01 99 Lot Time 10 55 07AM Operator Wavelength 405 Temp Plate 1D 456 COMMENTS RESULTS INVALID Control validation failed Interpretation of Results bl k_ave BLK x 20 176 GREYZONE 0 10 0 100 CUTOFF 1 1 000 SMP SMP lt CUTOFF CUTOFF GREY ZONE AND SMP gt CUTOFF CUTOFF GREVZONE EQUIV SMP SMP gt CUTOFF CUTOFFSGREYZONE POS SMP SHP lt CUTOFF CUTOFF GREYZONE NEG CTLE CTL1 x gt
18. 34074 39602 36231 Dark 9695 9743 9767 9718 9763 Delta 21750 28140 24307 29884 26468 Lambda 352 Gain 1 84 Resets 4 Channel Ref 1 2 3 4 Air 29680 37687 35488 38364 37300 Dark 9706 9730 9750 9716 9747 Delta 19974 27957 25738 28648 27553 Lambda 639 Gain 2 42 Resets 2 Channel Ref 1 2 3 4 Air 30309 39090 35718 39052 37655 Dark 9679 9749 9785 9712 9779 Delta 20630 29341 25933 29340 27876 Lambda 790 Gain 1 54 Resets 1 Channel Ref 1 2 3 4 Air 30435 39040 35751 39081 37564 Dark 9673 9766 9802 9721 9796 Delta 20762 29274 25949 29360 27768 Lambda 860 Gain 1 95 Resets 1 Channel Ref 1 2 3 4 Air 30306 38759 35633 38765 37221 Dark 9657 9775 9822 9719 9813 Delta 20649 28984 25811 29046 27408 Lambda 962 Gain 2 23 Resets 1 Channel Ref 1 2 3 4 Air 30088 38770 35545 38758 37241 Dark 9648 9780 9836 9716 9825 Delta 20440 28990 25709 29042 27416 Channel Ref 1 2 3 4 Noise Max 9634 9686 9775 9648 9766 Noise Min 9632 9684 9773 9647 9764 Delta 2 2 2 1 2 02 51PM 08 30 02 INCUBATOR SELF TEST Temperature Setpoint 37 0 Current Average Zone 1 36 9 Min 36 9 Max 37 0 Range Zone 2 37 0 Min 36 9 Max 37 1 Range Zone 3 36 9 Min 36 9 Max 37 1 Range Zone 4 37 0 Min 36 9 Max 37 1 Range SYSTEM TEST PASS Figure 4 1 Sample Output for the System Test PowerWaveX Operator s Manual 5 38224 9810 28414 38312 9781 28531 39196 9848 29348 39318 9871 29447 39047 9908 29139 39089 9934 2915
19. 40 000 Fc 60 000 70 000 CONCENTRATION Curve log y Ax 2 Bx C Coefficients A 000 B 0 027 C 743 R Sqr 0 9952 Figure D 2 Curve Fit Report showing the curve method equation coefficients and r squared value PowerWaveX Operator s Manual D 3 Bio Tek Instruments Assay ASSAY1 Date 07 01 99 Lot Time 10 36 10AM Operator Wavelength 405 Temp 30 C Plate 1D 123 COMMENTS Some concentrations may lie outside curve asymptotes lt gt around RSLT indicate extrapolated concentration Interpretation of Results bl k_ave BLK x 20 175 GREYZONE 0 10 0 100 CUTOFF 1 1 1 100 SMP SMP lt CUTOFF CUTOFF GREVZONE AND SMP gt CUTOFF CUTOFFFGREYZONE EQUIV SMP SMP gt CUTOFF CUTOFF GREYZONE POS SMP SMP lt CUTOFF CUTOFFXGREVZONE NEG Well ID 81xd0D Calc0D Call Pred Conc RSLT Std Dev CV Notes A01 BLK 0 031 0 031 24 629 gt Boi BLK 031 091 4 att AVE 0 000 0 000 lt 80 227 gt 0 044 error Col STD 0 290 0 290 lt 8 163 gt D01 STD 0 324 0 324 10 227 AVE 0 307 0 307 10 000 9 215 0 024 7 841 E01 STD2 0 526 0 526 20 136 F01 STD 0 557 0 557 21 416 AVE 0 541 0 541 20 000 20 781 0 022 4 023 601 STD3 0 818 0 818 31 012 HOt STD3 0 853 0 853 32 187 AVE 0 835 0 835 30 000 31 602 0 025 2 969 A02 STD4 1 089 089 39 786 B02 STD4 1 092 1 092 39 862 AVE 1 090 090 40 000 39 824 0 002 0 158 c02 STDS 1 324 324 47 244 D02 STD5 1 339 1 339 47 752 AVE 1 331 KK 50 0
20. E conte dedi E EA pita 3 33 Number of Required Blanks Controls ooooononccnninocinnnoncccnanannnononancnnnannncnnnn no 3 34 Assay Validation Formulas oooooooconnncccnnnoccccnonocnnononn coca nann nn nono cnn rar nn nn nana rra 3 34 EXAMPIl S iii dk 3 34 Transformation Formulas oooccnonccnnococcnnnonnnonanoncno nono ncnnnonn nn nano nn nr nan c nn anna rr 3 35 Simple Transformation Formulas oooooncconnccccnonoccnononannnononnncnnno conc nono cnn nn nnnnnnnos 3 35 Transformation Variable TVAR ooocconnnonicnnncoccnnnoncncnonannnnnnrncnnnn nn cc nnnonnnrnnnnnns 3 35 EXAMPpleS nt A a be Sasi hud Mee See eee 3 36 Cutoff Formulas iz avec ci aa 3 38 CTO ZOMBI es ei eo ait 3 38 POS NEG GallSiiuucitna tao viens 3 39 Example Sinn neti n 3 39 General Formulas acicate isis 3 40 Define CURVE iii acts 3 41 CUrVE Fit Type iii tance ania ideia aed adei a a iia 3 41 Edit Standard Outliers 0 eee eeeeeeeeeeeneeeeeeeeeeeeneeeeeeaeeeeeeaeeeenneeeenenaeeeeneaas 3 42 XINGAXIS TYPO OA 3 43 Extrapolation Of UNKNOWNS oooooccccnococccononcnonononcnn nono cnn nan nn n nano cnn rnnn nn n narran 3 43 Panel ASSays iactita dto 3 44 Reading a Microplate axiales 3 47 Selecting AN ASSAY caciones 3 48 Run Time Prompts3 ccn sen adele ai sheen ie Males ee clea ees 3 48 Enter Number of Samples a i 3 49 Enter Plate Dictada 3 49 Enter Sample Diciendo ee dacs 3 50 Enter Well Location eiert hiie ra neee eeraa tticcdse 3 50 Beginning the Plate
21. EEC for Low Voltage Verification of compliance was conducted to the limits and methods of the following e EN 61010 1 1993 Safety requirement for electrical equipment for measurement control and laboratory use Part 1 General requirements including amendment No 2 1995 viii Preface Safety Symbols The following warning and informational symbols may be found in various locations on the PowerWaveX Only qualified personnel who recognize shock hazards and are familiar with the safety precautions should use this instrument Read the manual carefully before operating this instrument ns Alternating current Courant alternatif Dreiphasen Wechselstrom Corriente Alterna Corrente alternata Earth ground terminal Borne de terre Erde Bettriebserde Borne de Tierra Terra di funzionamento Protective conductor terminal D Borne de terre de protection Schutzleiteranscluss Borne de Tierra de Protecci n Terra di protezione On Supply Marche alimentation Ein Verbindung mit dem Netz Conectado Chiuso Off Supply C Arr t alimentation Aus Trennung vom Netz Desconectado Aperto sconnessione dalla rete di alimentazione PowerWaveX Operator s Manual ix Caution risk of electric shock Attention risque de choc electrique Gefahrliche elektrische Spannung Atenci n riesgo de sacudida el ctrica Alta tensione in questo documento Alta tensione non significa tensione pericolosa come
22. ERR ABORT ERR NO SENSOR ERR NO BEAM ERR MOTOR VERIFY ERR SATURATION ERR FILTER GAIN ERR NOISE TEST ERR OFFSET TEST ERR DARK RANGE ERR AIR RANGE ERR ASSAY NUM ERR PRINT TIMEOUT ERR CAL CHECKSUM ERR WAVE NOT FOUND ERR FILTER SIGNAL ERR CNFG DATA ERR CNFG CHECKSUM ERR CAL DATA ERR MOTOR NOT HOMED ERR INCUBATOR FAILURE SC ASSAY DEF ERR KIN INTERVAL ERR KIN COUNT ERR MALLOC ERR STORE CURVE ERR 1A00 GET CURVE ERR ATOD INIT ERR A100 A200 A300 A400 A500 A600 A700 A800 A900 AA00 0000 0100 0200 0300 0400 0500 0600 0700 0800 0900 0A00 0B00 0C00 0D00 QE00 0F00 1000 1100 1200 1300 1500 1600 1700 1800 1900 task control block not available read already in progress lt device gt not available failed code checksum test on powerup DR steps alloc free error lt assay num data flash write timed out data flash readback didn t match write test lt chip gt code flash write timed out memory allocation heap corrupted lt device gt A D converter never saw ready transition no errors detected read function aborted lt motor gt didn t find opto sensor transition lt motor gt didn t find saturation transition lt motor gt failed posi
23. Edit Setup Information menu e Select DATE The DATE entry screen will appear pee Oe Aare re GEN ieee Entering the Date and selecting a format e Enter the date using the numeric keys on the keypad The cursor is positioned under the first editable field and advances automatically e To change the date format select MMDDYY or DDMMYY The display automatically updates to reflect the new format e Press ENTER to return to the Edit Setup Information menu e To change the current time and or the time format select TIME from the Edit Setup Information menu The Time entry screen will appear ARCO Ie ee ree ee Ih Wie E E Entering the Time and selecting a format e Enter the time using the numeric keys on the keypad The cursor is positioned under the first editable field and advances automatically e To change the time format select 12HOUR or 24HOUR then AM or PM The display automatically updates to reflect the new format e Press ENTER to return to the Select Utility Options menu Installation SPECIFYING DATA OUTPUT AND REPORTING OPTIONS To specify data output options e From the Main Menu select UTIL SETUP OUTPUT The Report Output screen will appear Data Output Selecting the Report Output e The current output option is displayed on the top line Select PRINT to send reports directly to a printer COMPUTER to send data out through the serial port or BOTH e Note
24. Manual 4 15 Procedure B Required Materials e Bio Tek QC Check Solution No 1 P N 7120779 25 ml or 7120782 125 ml e Deionized water e 5 ml Class A Volumetric Pipette e 100 ml Volumetric Flask Preparation of Stock Solution 1 Pipette a 5 ml aliquot of Bio Tek QC Check Solution No 1 into a 100 ml volumetric flask 2 Make up to 100 ml with DI water cap and shake well gt This should create a solution with an absorbance of about 2 000 when using 200 ul in a flat bottom microwell The OD value result will be proportional to the volume in the well and the amount of QC Check Solution No 1 used You can use a larger or smaller well volume or add more Check Solution or water to adjust the stock solution Note that too small a well volume may result in increased pipetting related errors Performance Verification Qualification Tests LIQUID TEST 1 This procedure will test for repeatability and consistency making evident any problems with the optics of the system 1 Using a freshly prepared stock solution see Procedure A on page 4 15 or Procedure B on page 4 16 prepare a 1 2 dilution using deionized water one part stock one part deionized water the resulting solution is a 1 2 dilution gt The concentrated stock solution will have an optical density of approximately 2 000 Abs This value is not critical but should be at the higher end of this absorbance range Should it exceed the range simply reduce
25. Port button and subsequent Setup button to define the following communications parameters e Port COM1 2 3 or 4 the serial port used for the RS 232 cable connection e Baud Rate 9600 e Data Bits 8 e Parity None e Stop Bits 2 Click the Current Reader button to attempt to establish communications with the reader using the currently defined communication parameters If the test passes click OK to save the settings and close the dialog box If the test fails KC4 will provide appropriate instructions for resolving any problems See also the Problems section below To learn more about KC4 and how to define assays and read plates refer to the KC4 User s Manual Problems B 2 If KC4 fails to communicate with the reader and it displays a serial communications error check the cable plug in location to ensure that it matches the setup choices and is not a Null cable If this is suspected add another Null and try again If an Incorrect Reader Model Connected dialog box is displayed click OK to clear the screen and select System Readers Available Readers Verify that the reader selected is correct Computer Control Table B 1 Serial Cable Pin Out Description Serial Cable Pin Out PC 9 pin female Reader 25 pin female 6 8 Shell Shell shielding For a 25 pin PC connection using Bio Tek serial cable PN 75053 plus 9pM 25pF adapter PN 49755 Getting Started with KC4 The fo
26. PowerWaveX see Figure 2 3 below allows connection to Epson and HP compatible deskjet printers See Compatible Printers on page 2 17 for information on specifying a printer The reader s parallel port requires a 25 pin D sub connector AL J Parallel port Monochromator viewing port Serial port Figure 2 3 Serial and parallel connectors The parallel port s pinout definition is described in Table 2 1 A printer cable PN 71072 is supplied with the reader If the cable that came with the reader gets lost or damaged Bio Tek offers replacement printer cables Contact your authorized Bio Tek dealer for information on cable prices and availability 2 6 Installation To attach a printer to the PowerWaveX Important To avoid system instability connect the printer to the reader before powering up the reader Turn on the PowerWaveX first then the printer Power down the printer and place it in a location adjacent to the PowerWaveX Attach one end of the parallel cable to the printer s parallel port Attach the other end of the cable to the reader s parallel port located on the instrument s rear panel see Figure 2 3 up the reader and printer Ensure that the securing screws on both ends of the cable are tightened and power PARALLEL PORT PIN DEFINITION Table 2 1 illustrates the pin definitions for the reader s 25 pin socket female D sub Parallel connector Table 2 1 Parallel Connector P
27. These options have no effect on data output if the reader is being controlled by software such as KC4 or KCjunior running on a host PC e Press ENTER to continue The Select Printer screen will appear Compatible printers Selecting a printer e BTI readers support printers using either HP s PCL3 language such as the HP DeskJet series or Epson s LQ language For the latest list of compatible printers contact Bio Tek Instruments Technical Assistance Center see page 1 8 for details or visit our website www biotek com e Select EPSON or HP as appropriate e Press ENTER to continue The Report Type screen will appear PowerWaveX Operator s Manual 2 17 Report Types Selecting the Report Type e Note Appendix D contains sample reports e The currently selected report type is displayed in the top line Select COLUMN to print information in a list columnar format MATRIX to print in a format that resembles the plate type e g 8 x 12 matrix or BOTH e Press ENTER to continue The Samples in Col Rpt screen will appear Specifying whether or not to include samples on the Column Report e Select YES to print results for all wells on the plate including samples e Select NO to limit the results information to blanks controls and standards e Press ENTER to continue The Print Curve Fit screen will appear Specifying whether or not to print the standard curve e Select YES to print the stand
28. as a model for calculating the expected absorbances of a series of dilutions given a different absorbance of the original solution PowerWaveX Operator s Manual 4 19 4 20 Linearity Test A Read the microplate prepared above using a normal mode dual wavelength at 450 nm with 630 nm as the blank Repeat the read four times for a total of five reads The plate data can be exported to an Excel spreadsheet using KC4 or KCjunior The mathematical computations described below may then be performed and the template kept for future data reduction Calculate the mean absorbance for each well and average the means for each concentration Perform a regression analysis on the data to determine if there is adequate linearity For example In Microsoft Excel under Tools select Data Analysis and then Regression Prior to opening the regression analysis tool enter the expected results of the dilutions in a row of cells to use in the analysis Use the Regression Input box to enter the expected values as the Input Y Range and the mean absorbance for each concentration as the Input X Range Expected results Since it is somewhat difficult to achieve high pipetting accuracy when conducting linear dilutions an R Square value of at least 0 99 is considered adequate Repeatability Test B Calculate the mean and standard deviation for the five readings taken above at each concentration Only one row of
29. backward through options The menu functions appear in the bottom second line of the front panel s LCD To make a selection press the SoftKey below the menu function SCROLLING THROUGH OPTIONS Certain functions such as entering an assay name or selecting an assay offer a set of options from which to make a selection To view the different options press the Options key or the Shift Options key combination Press the ENTER key to select the current option 3 2 Operation ENTERING ASSAY NAMES DNA Quant Each PowerWaveX assay requires the entry of a name using up to 16 alphanumeric characters To enter an assay name via the keypad press Shift key A H or scroll through the alphabet with the Options key for A Z Use the forward or reverse arrows to move the cursor within the display To delete an entry press the CLEAR key Press the ENTER key to store the completed assay name and continue The NAME display offers four symbols that can be used in an assay name dash hyphen forward slash colon and underscore These symbols appear in the LCD as SoftKey choices see above To include a symbol within an assay name press its corresponding SoftKey MOVING BACKWARD THROUGH THE MENUS To move to a previous menu press the Previous Screen key SAVING ASSAYS After defining an assay press the Main Menu key The changes are saved automatically STARTING OR STOPPING A READ To beg
30. character string with a null terminator 0x00 as the last character If the barcode is less than 32 characters it is null terminated then padded with blank characters 0x20 up to the final null If the barcode is more than 32 characters it is truncated If no barcode is detected after two passes of the microplate carrier in front of the scanner the null terminated string NR is returned followed by the remainder of the 33 character string of blanks and the final null A barcode error is flagged and returned with the standard status response string GET INSTRUMENT CONFIGURATION This command returns a 16 bit binary encoded word defining the instrument configuration host valid limits response y no arguments lt ACK NAK gt configuration 2 no status string Instrument configuration data is returned low byte first Encoding is defined as follows code description 0x0030 PowerWave 200 0x0031 PowerWave 340 0x0032 PowerWavex 0x0038 PowerWave 200 w Incubator 0x0039 PowerWave 340 w Incubator 0x003A PowerWave w Incubator 0x003B PowerWave 340 w Incubator 0x0072 PowerWavex Select 0x007A PowerWavex Select w Incubator 0x00B0 PowerWave 200 w Robot Access Door PowerWaveX Operator s Manual B 19 code 0x00B1 0x00B2 0x00B8 0x00B9 Ox00BA 0x00F2 Ox00FA description PowerWave 340 w Robot Access Door PowerWave w Robot Access Door PowerWave 200 w Incubator and Robotic Access Door Powe
31. complete and the results are not used in further data reduction Note General Formulas are not supported in the current version of the reader 3 40 Operation Define CURVE To define curve fitting parameters for an assay start at the Main Menu and select DEFINE select the assay press ENTER then select CURVE Selecting the Curve option on the DEFINE screen Configurable curve fitting parameters include e Curve Fit Type e Editing of Outliers e Axis Identification e Extrapolation of Unknowns CURVE FIT TYPE The PowerWaveX supports 7 different curve fitting methods Linear Quadratic Cubic 4 P 2 P cubic spline and point to point ESPE Ae PARE Selecting a curve fit type e NONE Choose None if no standard curve will be generated this is the default e LINEAR A simple best fit straight line is plotted using the values of the standards PowerWaveX Operator s Manual 3 41 QUADratic Uses the Quadratic equation ax bx c y to plot the standards values Utilizing this curve any data point for a standard that deviates from the ideal value will not affect the entire curve CUBIC Uses the equation ax bx cx d y to plot the standards values This type of curve fit is affected even less than the quadratic fit when any particular standard has a poor value 2 P Logit Log A curve fitted to the standard values which is characterized by a skewed sigmoidal
32. data needs to be analyzed For each mean below 2 000 Abs calculate the allowed deviation using the repeatability specification for a 96 well format of 1 0 005 Abs If above 2 000 Abs apply the 3 specification The standard deviation for each set of readings should be less than the allowed deviation For example Absorbance readings of 1 950 1 948 1 955 1 952 and 1 950 will result in a mean of 1 951 and a standard deviation of 0 0026 The mean 1 951 multiplied by 1 1 951 0 010 0 0195 which when added to the 0 005 0 0195 0 005 0 0245 Abs which is the allowable deviation Since the standard deviation is less than this value the reader meets the test criteria Performance Verification Qualification Tests Repeatability Specification 1 0 005 Abs from 0 000 to 2 000 Abs 3 0 005 Abs from 2 000 Abs to 3 000 Abs Channel to Channel Variation and Alignment Test C 1 Using the plate prepared for Test A above conduct a turnaround test by reading the plate with the Al well in the H12 position five times This test results in four comparisons of each channel to its corresponding channel two in column 1 and two in column 2 2 Calculate the means of the wells in columns and 2 in the normal plate position data is from Test A and in the turnaround position from Step 1 above Compare the mean reading for well A1 to its mean reading when in the H12 position Nex
33. difference in the values for any two corresponding wells should be within the accuracy specification for the instrument PowerWaveX Operator s Manual 4 17 For example If the mean of well A1 in the normal position is 1 902 where the specified accuracy is 1 0 010 Abs then the expected range for the mean of the same well in the H12 position is 1 873 to 1 931 Abs 1 902 1 0 019 0 010 0 029 which is added and subtracted from 1 902 for the range If any set of well values is out of the expected range review the other three sets for the same channel pair Thus if Al and H12 are not within range of each other review the compliance of H1 to A12 A2 to H11 and H2 to A11 This will confirm that there is a problem in one of the eight read channels or indicate that the result of one set of wells was in error If two or more sets of well values for a channel pair are out of the allowed accuracy range there may be contamination on or a problem with one of the lenses Accuracy Specification For comparison in this test the following accuracy specifications are applied using Normal mode and a 96 well microplate 1 0 010 Abs from 0 000 to 2 000 Abs 3 0 010 Abs from 2 000 Abs to 3 000 Abs Performance Verification Qualification Tests LIQUID TEST 2 The recommended method of testing the instrument performance is to use the Universal Test Plate to confi
34. is rated at an average of 1 billion flashes This bulb should outlive the useful life of the reader If there is a problem with the lamp however the intensity may drop and the run time self check will detect a low signal level and generate an error message If this happens contact Bio Tek for bulb replacement information COOLING FAN Located on the bottom right edge of the instrument is a switch that activates an internal cooling fan When the PowerWaveX is on the temperature in the measurement chamber will be approximately 7 C above ambient temperature For users that need the measurement chamber to be close to ambient Bio Tek recommends using the cooling fan The fan will bring the temperature within 4 C of ambient This would be important for example for kinetic measurements that need to be made close to room temperature The fan should be off when non temperature sensitive testing is being completed as prolonged use may introduce dust into the internal mechanisms of the instrument PowerWaveX Operator s Manual 2 11 Adjusting the Line Voltage Input Range The PowerWaveX is equipped with a four voltage range power input module This power input module located on the right side of the instrument can be adjusted for 100 VAC 115 VAC 230 VAC and 240 VAC voltage inputs The setting can be determined visually by observing which indicator hole on the power input module has a peg within it The ON OFF switch and fuses are al
35. mapped in each individual assay while mapping for the Panel assay For example to include Assays 1 8 and 22 in the Panel assay DOWN mapping is selected for the Panel gt Assay 1 has a total of 12 wells defined for controls blanks and standards In the Panel the mapping for Assay 1 begins in well A01 The user wants to run 6 samples in Assay 1 Assay 1 now fills wells A01 through B03 gt The mapping for Assay 8 can begin in well B04 or any well other than A01 to B03 The reader will beep if you try to map into a well that is already assigned for use with the Panel gt The mapping for Assay 22 may begin at the next available well location after Assay 8 mapping is complete gt After all the assays have been entered into the Panel consider printing the Panel s Map Report to verify the map before reading the plate Choose REPORT from the Main Menu MAP ASSAY 99 The reader will print the map of each assay configured in the Panel gt The Panel Assay results are sorted by Sample unless a custom assay has been programmed by Bio Tek Note The Interpretation of Results reports for each assay in the panel will print first and then the Sample results will print See Appendix D for a sample Panel Report 3 46 Operation Reading a Microplate Use the Main Menu option READ or press the READ key on the front panel to initiate a plate read REPORT UTIL ECT ASSAY NUMBER
36. of each type are shown as well Blank Validation 0 1 Control Validation 0 4 Assay Validation 0 4 Transformations 0 1 Cutoff Formulas 0 1 Curve Fit Analysis if a curve fit method is defined General Formulas 0 4 PowerWaveX Operator s Manual 3 29 FORMULA TYPE 3 30 The PowerWaveX supports four types of formulas as well as the ability to program variables for use within Transformation formulas Selecting the type of formula to create or edit e CUTOFF formulas are used to classify results During data reduction results are evaluated against the cutoff formulas and each well is assigned a user specified label POS NEG or EQUIV e TRANSformation formulas are applied to the absorbance data in preparation for further data reduction and or curve fit calculation e VALidation formulas can be used to determine whether or not blanks and or controls are valid In addition Assay Validation formulas can be used to determine whether or not the entire assay should be considered valid e GENERAL formulas are calculated after all other calculations are complete and the results are not used in further data reduction Note General formulas are not currently supported in the PowerWaveX on board software e The TRANS VAR option allows you to define a variable to be used in transformation formulas Operation FORMULA ENTRY After the formula type is maximum of 24 characte selected the Form
37. operations e Ability to define controls and positive and negative cutoffs e Kinetic read capability with maximum slope R squared at maximum slope and Onset OD Time analysis e Ability to perform a spectral scan reading from 200 to 999 nm or 340 to 999 nm depending on the model e Scanning read mode to provide area under the curve calculations e Optional preprogrammed common biological assays Specimen Preparation Samples should be obtained treated and stored following instructions and recommendations determined by the user s laboratory Quality Control It is considered good laboratory practice to run laboratory samples according to instructions and specific recommendations included in the package insert or standard laboratory protocol for the test to be conducted Failure to conduct Quality Control checks could result in erroneous test data PowerWaveX Operator s Manual xi About This Manual The intent of this Operator s Manual is to quickly instruct the new user how to set up and operate Bio Tek s PowerWaveX To help you read and understand this manual certain document conventions have been used Major topic headings start a new page such as About This Manual above to give you a visual and style clue that a new major subject is being introduced One or more subheadings may appear below each major heading Registration Card Once the PowerWaveX has been set up and is running successfully please take a mom
38. sheet BTI P N 7262113 is in place on top of the main PCB and below the motor assembly before lubricating the motor Tape to the chassis with black electrical tape This may already be in place E Lightly lubricate the surface of the roller F Lightly lubricate the underside of the hook in bracket 3 Reassemble the shroud 4 After lubrication reassembly the robotic door needs to be set to the open and closed positions 5 Press the following keys to adjust the door At the Main Menu Press the UTIL soft key Press the Setup soft key Press the hidden key between the Main Menu and Previous Screen keys 6 You should see the Door Adjustment screen 7 Press the CLOSE soft key PowerWaveX Operator s Manual 4 25 8 Press the UP soft key until you just see the door move up You may have to press the key many times before the door moves When the door does move press the DOWN soft key 4 times This positions the door lift ram just behind the door roller in the closed position 9 Press the OPEN soft key 10 Press the UP soft key to raise the door to the desired height Press the DOWN soft key if you want to adjust the door downward 11 Press the CLOSE soft key This will close the door The door should rest closed on the top shroud 12 Press the Main Menu key 4 26 Performance Verification Qualification Tests Figure 4 5 Location of bracket motor shaft mylar sheet and main PCB motor assembly 4 27 Pow
39. such All calculated results for an Ambiguous curve should be considered unreliable PowerWaveX Operator s Manual 3 43 Panel Assays A Panel assay is a collection of up to eight assays to be run on one plate The most common reason to use a Panel assay is for confirmatory tests based on a screening test in clinical applications Only one panel assay can be defined on the reader at any time The assays specified within the Panel must be pre defined in any of the assay positions 1 55 The assays specified within the panel must all use the Endpoint read method The assays specified within the panel must all read at the same wavelength s Any curve fit type formulas or standard concentrations previously defined for each assay will be used when the assay is selected for a Panel The type and number of controls blanks standards and replicates in the assays chosen for the Panel will be copied into the Panel definition To change any of the map or assay parameters in the Panel they must be changed in the pre defined assay first Consider printing a Map Report for each assay that will be included in the panel for use with mapping the Panel To create a panel assay start at the Main Menu press DEFINE then choose assay number 99 Enter the panel assay name ISA AE Editing the name of the Panel assay 3 44 The default name is PANEL Use the ALPHA and NUMERIC keys to update the Assay name i
40. the temperature from 4 above ambient to 50 C e R model instruments are robotic compatible e An internal bar code scanner option is also available 111 Note 384 well plate reading must be performed through computer control or with KC4 or KCjunior software See Appendix B for more information Hardware Features Xenon flash light source Eight optics channels with an additional reference channel Monochromator with A range of 200 to 999 nm or 340 to 999 nm depending on reader model A 2 X 24 character LCD display A membrane keypad with alphanumeric keys Adjustable plate shake frequency and durations Reads 96 well microplates with 0 355 9 mm well centers Reads 384 well microplates with 0 177 4 5 mm centers PowerWaveX Select model only Operates from 100 120 230 or 240 VAC 50 60 Hz with external switching One serial COM port 25 pin male connector One parallel port 25 pin female connector Internal bar code option 4 Zone incubation chamber option Robotics interface option Software Features 1 2 Easy to use menu driven interface Endpoint Kinetic and Scanning calculations Curve fitting with 4 parameter cubic quadratic linear 2 P cubic spline and point to point methods Transformation and formula calculations for more complex mathematical operations including validations Up to 55 assays are available on board 27 open assays plus preprogrammed common bio
41. the above example e After each entry press ENTER to advance to the next consecutive well location e When the sixth value has been entered press ENTER to return to the Calibration Lambda selection screen to select another wavelength or Main Menu Running the Universal Plate Test Before running the Universal Plate Test ensure that the standard calibration values for the test plate have been defined on the reader see Entering the Universal Test Plate Data on the previous page To run the Universal Plate Test e Ensure that the printer is attached turned on and full of paper e From the reader s Main Menu press UTIL TESTS CAL PLATE The Calibration Lambda selection screen will appear pie ele ee e aa Selecting a wavelength for the calibration plate test e Select a wavelength for the test then press ENTER PowerWaveX Operator s Manual 4 9 The Place Plate in Carrier prompt will appear Place the Universal Test Plate in the reader then press READ e Place the Universal Test Plate in the carrier so that well A1 is in the upper left corner of the carrier then press READ to begin the test The plate will be read twice then the Rotate Plate prompt will appear Beep aaa Jaye E Rotate the plate then press READ e Rotate the plate so that well A1 is in the lower right corner of the carrier then press READ to complete the test e When the test is complete results will print See the sample Cal
42. the next standard Number of Controls For assays requiring controls begin by entering the number of control groups Entering the number of control groups e Use the NUMERIC keys to enter the Number of Control groups in the assay For example if the assay requires one or more positive control wells and one or more negative control wells enter 02 e The valid entry range depends on the number of locations on the plate that are undefined The maximum number of control groups is 8 PowerWaveX Operator s Manual 3 25 Control Type After defining the number of controls for this assay select the types of controls to use eee ere ee ee E piles PTS Aaa eee Cea E Seale eee eee alee ESRC ERERE RC ON RARER Selecting the types of controls to use with this assay e Choose one control identifier for each type of control in your assay The available options are Positive Control Negative Control High Positive Control Low Positive Control CTL1 CTL2 CTL3 CTL4 e After choosing an identifier for Control 1 press ENTER to choose the identifier for the next control Number of Control Replicates The Number of Control Replicates entry screen is presented if the number of control groups is greater than 0 Be Sie le eee ee Wale Entering the number of PC Positive Control replicates e The well ID associated with Control 1 appears first Press ENTER to advance to the next control e Use the NUMERI
43. to determine the minimum kinetic read interval as dictated by the current assay definition table strip range and even baud rate selected host valid limits response no arguments lt ACK NAK gt kinetic interval 2 status string 5 The reader returns the minimum kinetic interval as a 2 byte integer value not ASCII so low byte first PowerWaveX Operator s Manual B 13 SCAN PLATE amp This command causes the reader to perform a spectral scan on the indicated microwell host amp data bytes 1 2 15 16 17 18 lt ETX gt B 14 valid limits response lt ACK NAK gt row 00 16 ASCII 00 indicates scan entire strip column Note Do not use 00 if reading 384 well plate column 01 24 ASCI start wave 200 999 ASCID stop wave 201 999 ASCII wave step 001 799 ASCII series option 0 3 ASCII 0 single scan 1 first scan in series 2 next scan 3 last scan calibrate option 0 1 ASCID 0 calibrate only if necessary I calibrate before scanning shake time in seconds 00 99 ASCII shake speed 0 slow medium 2 fast 3 cycle through speeds listed above lt ACK NAK gt Computer Control host valid limits response Reader response protocol ASCII format status string 5 for each wavelength undergoing cali
44. 00 47 498 0 011 0 824 E02 STD6 1 619 619 57 917 F02 STD6 1 610 1 610 57 558 AVE 1 614 614 60 000 57 737 0 006 0 373 602 STD7 1 873 873 e H02 STD7 1 914 1 914 d 34 AVE 1 894 894 70 000 4 dk 0 029 1 540 Page 1 Figure D 3 First page of Column Report showing plate data for blanks and Standards the Interpretation of Results and comments D 4 Sample Reports Well 10 A03 B03 AVE c03 D03 AVE E03 F03 AVE G03 H03 AVE A04 B04 AVE C04 D04 AVE E04 F04 AVE G04 H04 AVE A05 B05 AVE 05 DOS AVE E05 FOS AVE 605 H05 AVE A06 806 AVE C06 Page 2 PowerWaveX Operator s Manual SHPOOO1 SHP0001 SHP0002 SMP0002 SMP0003 SHPO003 SMP0004 SMP0004 SMP0005 SMP0005 SMP0006 SMP0006 SHMP0007 SMPOOC SMP0008 SMP0008 SMP0009 SMP0009 SMP0010 SHP0010 SMP0011 SMP0011 SMP0012 SMP0012 SMP0013 SMP0013 SMPOG14 BIkdOD lt gt oo a as ya o o gt gt 0 0 0 A a ae oo 332 314 323 151 808 179 334 321 328 ATT 758 168 057 144 104 825 819 822 1095 080 087 1275 248 262 779 807 793 047 320 184 644 816 130 1519 495 507 509 672 590 148 CalcOD Doc gt se oso gt Doo Oo 332 314 323 151 908 179 334 321 328 J1 758 188 087 144 101 825 819 822 095 080 087 278 248 262 179 807 793 047
45. 05 E DNA Quant Press OPTIONS to scroll through the assay list Press ENTER to select an assay PLATE ID 45768 A PLATE IN CARRIER ESS lt READ gt KEY Figure 3 5 Options available under READ Notes The options to present the Enter Number of Samples Plate ID and or Sample ID screens are configurable via UTIL gt READ Custom assays may present additional screens for the entry of special parameters refer to the Assay Reference Guide PN 7271006 for more information Notes Before reading a plate ensure that the reporting options are set correctly under UTIL Y OUTPUT See Selecting Read Options in Chapter 2 for more information PowerWaveX Operator s Manual 3 47 SELECTING AN ASSAY From the Main Menu press READ and then select the desired assay OIDO Selecting an assay to run e Use the NUMERIC keys to enter the number of any predefined assay stored in the reader s memory or the OPTION key to advance one assay at a time e Press ENTER to continue RUN TIME PROMPTS After the assay is selected one or more informational prompts may be presented depending on preferences selected via UTIL gt READ whether or not the assay specifies manual mapping or if this is a custom pre programmed assay e Prompts enabled via UTIL gt READ can include Enter Number of Samples Plate ID and Enter Sample ID e Ifthe as
46. 29 99 Time 03 31 17PM Temp Calc0D 729 Za ss A DO OA gt a mR 382 A719 T44 13 206 387 326 173 190 184 a OD 069 090 080 606 626 616 205 124 163 SD o 1 367 1 076 1 222 2 463 2 538 2 502 Figure D 6 Panel Report Call NEG NEG NEG POS POS POS NEG NEG NEG NEG NEG NEG NEG NEG NEG POS POS POS NEG NEG NEG NEG NEG NEG NEG NEG NEG NEG NEG NEG POS POS POS NEG NEG NEG NEG NEG NEG POS POS POS RSLT Lot Operator Plate 1D 123456 Std Dev 0 014 0 023 0 029 0 046 0 022 0 024 0 017 0 043 0 012 0 015 0 014 0 060 0 206 0 052 Cv 1 889 1 399 3 742 2 653 1 409 2 392 3 248 1 025 1 391 0 542 5 133 16 869 2 068 Notes 10 48AM 07 01 99 NAME ASSAY2 READ TYPE ENDPOINT WAVELENGTH 405 630 BLANK MAP FULL AVERAGE STANDARDS YES CONCN STD1 0 0 STD2 2 0 STD3 5 0 STD4 10 0 STDS 20 0 STD6 50 0 CURVE FIT TYPE 4 P EDIT STD OUTLIERS MANUAL X Y AXIS TYPE LIN EXTRAPOLATE UNKNOWNS YES 1 2 3 4 5 6 7 8 9 10 11 12 BLK BLK SMP SMP SMP SMP SMP SMP SMP SMP SMP SMP STD1 STD1 SMP SMP SMP SMP SMP SMP SMP SMP SMP SMP STD2 STD2 SMP SMP SMP SMP SMP SMP SMP SMP SMP SMP STD3 STD3 SMP SMP SMP SMP SMP SMP SMP SMP SMP SMP STD4 STD4 SMP SMP SMP SMP SMP SMP SMP SMP SMP SMP STD5 STDS SMP SMP SMP SMP SMP SMP SMP SMP SMP SMP STD6 STD6 SMP SMP SMP SMP SMP SM
47. 4 1 144 0 807 0 672 1 347 0 816 0 820 1 392 0 754 1 050 Well BLK S104 SMP1 SMPS SMP SHPI3 SMP17 SHP2 1 SMP25 SMP29 SMP33 SMP37 RSLT ek 30 862 46 914 1 41 505 30 651 25 899 48 011 30 946 31 103 49 495 28 813 38 562 E __ _n __ _ __ OOooo _ om m CALL NEG NEG EQUIV EQUIV NEG EQUIV POS NEG POS NEG CalcOD 0 290 1 324 0 751 0 825 1 047 1 148 0 804 1 073 1 307 0 975 1 334 0 775 Well STM STDS SHP SMP6 SMP10 SMP14 SMP18 SMP22 SMP26 SMP30 SMP34 SHP38 RSLT lt 8 163 gt 47 244 28 703 31 254 38 467 41 629 30 560 39 272 46 697 36 152 47 584 29 557 A _ _a ____JJJ J __ A _ _J _ 0000000 o _ _ 0 _ CALL NEG NEG POS POS NEG EQUIV POS NEG POS NEG CalcOD 0 324 1 339 0 808 0 819 1 320 1 236 0 777 1 036 1 486 0 966 1 320 0 792 Well STOI STDS SMP2 SMP6 SMP10 SMP 14 SMP 18 SMP22 SMP26 SMP30 SMP34 SHP38 RSLT 10 227 47 752 30 670 31 070 47 118 44 432 29 640 38 108 52 758 35 864 47 131 30 125 Eg af __ 2 CALL POS EQUIV POS POS POS NEG POS EQUIV NEG EQUIV CalcOD 0 526 1 619 1 334 1 095 1 644 2 479 1 323 0 815 1 288 1 001 0 779 1 031 Well TD2 5TD6 SMP SMP SMP tt SMP15 SMP19 SHP23 SMP27 SHP31 SMP35 SMP39 RSET 1 20 136 57 917 47 582 39 966 59 017 4 438 47 224 30 915 46 086 36 993 29 675 37 945 F CALL POS EQUIV POS POS POS NEG POS NEG NEG EQUIV CalcOD 0 557 1 610 1 321 1
48. 5 9902 9899 36 9 PASS PASS PASS PASS 6 7 36345 36493 9819 9808 26526 26685 6 7 39652 36991 9786 9778 29866 27213 6 7 39901 38448 9861 9845 30040 28603 6 7 39895 38492 9889 9870 30006 28622 6 7 39715 38223 9932 9907 29783 28316 6 7 39940 38235 9961 9931 29979 28304 6 7 9915 9888 9910 9884 5 4 A D Test Thermistor Thermistor Thermistor Thermistor 8 39331 9718 29613 37723 9717 28006 38423 9712 28711 38449 9718 28731 38136 9714 28422 38095 9712 28383 8 9658 9656 PASS PASS PASS PASS PASS 4 5 CHECKSUM TEST CHKSUM The Checksum test runs automatically when the reader is powered on The test compares the software to the internally recorded checksum values to ensure that the programming has not been corrupted If there are any errors during the power on checksum test they will be displayed To verify the checksum manually and to view the part numbers and versions of software currently loaded onto your reader start at the Main Menu and press UTIL gt TESTS Y CHKSUM The information displayed will resemble the following the actual checksum will be dependent upon the software version that is loaded EEEE EE LEOTE REEE EECE et eee Eeee Ee The initial checksum test display showing the basecode software part number version number and checksum After a few moments a second screen will display ee DEE A e alll The second checksum tes
49. 60 Hz Range4 240VAC 216 264 VAC 50 60 Hz Temperature Control 4 over ambient to 50 C Temperature Variation 0 5 C across the plate 37 C with the plate sealed If enabled the internal bar code reader recognizes a number of common bar code types during the plate read operation Specific information about each bar code type is available from Bio Tek Technical Services The reader s bar code option is compatible with the following bar code types CODABAR UPC CODE 39 EAN INTERLEAVED 2 of 5 MSI CODE 11 PLESSEY CODE 93 CODE 128 The Robotic interface option allows the reader to function with an autoloading robot Using computer control commands from a host PC the reader s functions can be controlled in conjunction with the robotic system The Robotic interface model can be configured with all options available for the standard PowerWaveX 1 7 Technical Support 1 8 Bio Tek s PowerWaveX is backed by a superior support staff If the PowerWaveX ever fails to work perfectly please contact Bio Tek s Technical Assistance Center Whichever method of contact you choose please be prepared to provide the following information e Product name and serial number e The reader s on board software configuration information To see this start at the reader main menu and press UTIL Y TESTS CHKSUM e The specific steps that produce your problem e Any error codes displayed on the screen see Appendix C for in
50. C keys to enter a value for Number of Control Replicates e The valid entry range is from 1 to 12 replicates The software automatically performs a check to ensure the number of replicates multiplied by the number of controls does not exceed the number of undefined wells remaining on the plate 3 26 Operation Location of Controls If mapping is manual and controls are defined the locations for each control replicate must be specified SPE eee MUS eae BEEBE EMRE oda Entering the control locations if manual map e Use the NUMERIC and ALPHA keys to enter the well location for Rep 1 of Control 1 Press ENTER to advance to the next replicate or control group Number of Samples The number of sample groups on the plate can be defined here and or it can be defined at run time if UTIL READ PROMPT FOR SAMPLE COUNT is set to YES See Selecting Read Options in Chapter 2 for more information Entering the number of sample groups on the plate e Use the NUMERIC keys to enter the number of sample groups on the plate e The valid entry range is from 01 to the number of undefined well locations remaining on the plate e Ifthe Number of Samples setting is altered the Number of Sample Replicates setting resets to 01 Number of Sample Replicates After the number of sample groups is specified the Number of Sample Replicates entry screen is presented SAVE PEO Ida Entering the Number of Sample
51. DowerWave x Pe O BIO TEK INSTRUMENTS INC OH Microplate Scanning Spectrophotometer Operator s Manual Creating Solutions for the Future of Science 7261014 Rev D2 July 2005 Page 1 of 2 This Manual Update contains changes to the PowerWaveX Operator s Manual These changes will be incorporated in the next full revision Rev E of the Manual Page 4 17 Liquid Test 1 Recommendation After pipetting the diluted test solution into the microplate step 3 and before reading the plate we strongly recommend shaking the plate at Variable speed for four minutes This will allow any air bubbles in the solution to settle and the meniscus to stabilize Alternatively wait 20 minutes after pipetting the diluted test solution before reading the plate Pages 4 7 through 4 13 Universal Plate Test Clarification Bio Tek s Absorbance Test Plate part number 7260522 can be used to test the alignment accuracy repeatability and linearity of your PowerWaveX Microplate Spectrophotometer Revision D of the Operator s Manual refers to this test plate by its former name the Universal Test Plate BI1O TEK BIO TEK INSTRUMENTS INC P O BOX 998 HIGHLAND PARK WINOOSKI VERMONT 05404 USA COPYRIGHT 2005 TEL 888 451 5171 FAX 802 655 7941 Outside the USA 802 655 4740 E mail customercare biotek com www biotek com Creating Solutions for the Future of Science 7261014 Rev D2 July 2005 Page
52. E NEG SMP lt CUTOFF CUTOFF GREYZONE Examples 1 The cutoff between negative and positive calls should be calculated as the average of the negative controls plus the OD value 0 050 Samples greater than the cutoff should be labeled as positive No greyzone is required e For this example NC x the mean of the NC wells equals 0 100 OD e The cutoff formula is NC x 0 050 e The greyzone is 00 e POS is selected for SAMP gt CUTOFF 00 e Calls are assigned to sample wells as follows gt EQUIV if the sample equals 0 150 gt POS if the sample is greater than 0 150 gt NEG if the sample is less than 0 150 PowerWaveX Operator s Manual 2 Fora quantitative assay samples with ODs greater than the STD2 mean plus a 10 greyzone should be labeled as positive samples with ODs less than the STD2 mean minus the 10 greyzone should be labeled as negative All other samples should be considered equivocal e For this example STD2 x the mean of the STD2 wells equals 1 500 OD e The cutoff formula is simply STD2 x e The greyzone is 10 e POS is selected for SAMP gt CUTOFF 10 e Calls are assigned to sample wells as follows gt EQUIV if the sample is greater than or equal to 1 350 and less than or equal to 1 650 gt POS ifthe sample is greater than 1 650 gt NEG if the sample is less than 1 350 GENERAL FORMULAS General Formulas are calculated after all other calculations are
53. INE Text in COURIER font represents menu options as they appear in the instrument s display Note Bold text is primarily used for emphasis This icon calls attention to important information p o jo pmo iv Preface Warnings The PowerWaveX should be operated on a flat surface away from direct sunlight or strong incandescent light Excessive humidity should be avoided Please read the following hazards and cautions before operating the instrument HAZARDS Warning Power Rating The PowerWaveX must be connected to a power receptacle that provides voltage and current within the specified rating for the system Use of an incompatible power receptacle may produce electrical shock and fire hazards Warning Internal Voltage Always turn off the power switch and unplug the power cord before cleaning the instrument s outer surface Warning Liquids Avoid spilling liquids on the reader fluid seepage into internal components creates a potential shock hazard Do not operate the instrument if internal components are exposed to fluid Warning Software The microplate reader operator must follow the manufacturer s assay package insert when modifying software parameters and establishing result calculation methods using the reader s on board software Warning Data Reduction Protocol The reader s software will flag properly defined controls when they are out of range It will present all the data with the appr
54. L EE ee ete Eo e e The Main Menu e The Select Utility Option menu appears Selecting SETUP from the Utility Option menu Installation e From the Select Utility Option menu select SETUP The Edit Setup Information menu appears Selecting LAMBDA from the Edit Setup Information menu e From the Edit Setup Information menu select LAMBDA e The Enter Lambda 1 screen appears showing the first value in the wavelength table POR erat Viewing or changing the reader s internal wavelength table e To advance to the next wavelength press ENTER e To change the current wavelength value use the reader s numeric keypad to enter a number at the current cursor location Once a selection is made the cursor automatically advances to the next editable location To save the entry and advance to the next wavelength press the ENTER key e Any wavelength in the range 200 to 999 nm may be entered or 340 to 999 nm on the PowerWaveX 340 e When the last sixth wavelength has been entered or viewed the software returns to the Edit Setup Information menu Edit Setup Information menu e Press the Main Menu key to return to the Main Menu Note After selecting new wavelengths the reader system test must be run From the Main Menu press UTIL gt TESTS Y SYSTEM to start the test PowerWaveX Operator s Manual 2 5 Printing and Data Communications INSTALLING A PRINTER The parallel port on the
55. N Entering the Onset OD for kinetics e Use NUMERIC keys to enter the onset OD The valid entry range is 0 000 to 3 000 OD WELL SCANNING For the Scanning Read Type multiple readings are taken across each microwell The number of readings or Scan Points is configurable eS Peel eee E Entering the number of scan points per well e The maximum number of Scan Points is 31 Only odd integers are accepted to ensure that the center of the well is always read e The 31 scan positions are fixed in the software You must determine the optimal number of scans per well If for example 7 scans across the well is chosen the reader will read the centermost seven points in the well The more scan points chosen the closer to the well sides reads will be taken e The reader reports the sum of OD values for all points scanned Note Exercise caution when specifying the Scan Points value If too many points are specified the sides of the wells may be getting read possibly resulting in an artificially high reading 3 16 Operation Define MAP To configure the plate layout start at the Main Menu and select DE ENTER then press MAP E select an assay press Selecting the Map option on the DEFINE screen The following MAP parameters can be defined modified e Automatic or Manual Map Generation e Mapping Direction e Replication Direction e Blank Map Selection e Blanking Constant e Number of B
56. N j oh Y 0 ola oia ls N N j A jw A O w I a Figure 4 2 Sample Universal Test Plate data sheet Entering the Universal Test Plate Data Note Before defining the standard calibration values set the reader s internal pio pio pio wavelength settings to correspond with up to six of the wavelengths shown on the Universal Test Plate Data Sheet see Figure 4 2 above See Adjusting the Reader s Wavelength Table Settings in Chapter 2 for instructions Additional wavelengths above 405 nm are available at extra cost Contact your Bio Tek dealer Refer to the Data Sheet provided with the Universal Test Plate Figure 4 2 when entering the standard values To enter the values into the reader start at the Main Menu and press UTIL gt SETUP The Edit Setup Information menu will appear Edit Setup Information menu Press MORE to see the Cal Plate option e From the Edit Setup Information menu press MORE then select CAL PLATE Performance Verification Qualification Tests The Calibration Lambda entry screen will appear dao O OSO OEM IEA NANA DAA Selecting a wavelength for which to enter standard calibration values e Choose a wavelength e g 405 then press ENTER Entering standard calibration values referencing the Universal Test Plate Data Sheet e Enter the absorbance value from the Data Sheet for the selected wavelength and current well location C01 in
57. P SMP SMP SMP SMP SMP SMP SMP SMP SMP SMP SMP SMP SMP SMP SMP SMP TO0mMMOO CU Figure D 7 Assay Detail Report D 8 Sample Reports Assay List Version 2 0 01 Raw_OD 02 Raw_OD w blk 03 Bio Cell Read 04 K Factor Bio Cell 05 DNA Quant 06 260 280 Ratio 07 Protein Quant 08 Oligo Quant 09 Protein Quant 10 Phenol 11 EDTA test 12 A320 Scattering 13 Lowry Protein 14 Bradford Protein 15 BCA Protein 16 B galactosidase 17 NADH 18 HRP 5AS 19 HRP ABTS 20 HRP OPD S 21 HRP OPD 22 HRP TMB S 23 HRP TMB 24 Alk phosphatase 25 B galact ONPG 26 Urease 27 MTT 28 XTT 29 WST 1 30 Methylene Blue 31 Open assay 01 32 Open assay 02 33 Open assay 03 34 Open assay 04 35 Open assay 05 36 Open assay 06 37 Open assay 07 38 Open assay 08 39 Open assay 09 40 Open assay 10 41 Open assay 11 42 Open assay 12 43 Open assay 13 44 Open assay 14 45 Open assay 15 46 Open assay 16 47 Open assay 17 48 Open assay 18 49 Open assay 19 50 Open assay 20 51 Open assay 21 52 Open assay 22 53 Open assay 23 54 Open assay 24 55 Open assay 25 Figure D 8 Assay List PowerWaveX Operator s Manual Sample Reports
58. Readd ciicccinionicoc ad lidia 3 51 xviii Preface Printing Reports ui iaa 3 52 Results Repor er de chil es E Ra 3 53 Editing Standard Outliers oooooooococcccnnconocococccnccnoconanoncnnccncnnnnnnonnnnnnnnannnnns 3 53 Printing ResUlS cria 3 54 Map Reportan is 3 55 Assay Repito ea 3 55 List Repor naa A 3 55 Chapter 4 Performance Verification Qualification Tests 00 4 1 Recommendations for Achieving Optimum Performance ccccceeeeseeeeeeeeeeenneeeeeeneees 4 1 Recommended Test Schedule ooococcconococccononcnononananononnnonano cnn crono cnn nn n rra rr 4 2 Installation and Performance Qualification Procedures oooooccccnociconoccccnononcnnnnnannnnnnnnnn 4 2 Routine Procedure ic 4 3 System Testing a aba 4 4 NA caceeaua yh cusbstceacnadsoecunndscasebensshlusecbcs sueantebeashas cnsebeiges antes 4 4 Xenon Flash Lamp s cs ese cs li eee ek en ie a a eee 4 4 Incucai 4 4 Checksum Test CHKSUM ocoooiicococcion caco tenia 4 6 Calibration Verification dr 4 7 Universal Plate Testo sched seine vee 4 7 Requirements dela 4 8 Entering the Universal Test Plate Data oooonooocccnncnininocococcnonocanononcnoncnncnnnnn nono 4 8 Running the Universal Plate TeSt oooooocococnninococcccccnccocononccnncnocanano nono ncnnnnnannncnnn 4 9 Wavelength ACCUIACY diiniita 4 13 Liquid Testing iii ic 4 14 Stock Solution FormulatiON oononcccnnnnciccnnnocnnnnococcnanoncn crono nn n rro nnn nn nn rra rn 4 15 Proce
59. Replicates PowerWaveX Operator s Manual 3 27 e Use the NUMERIC keys to enter the number of sample replicates e The valid entry range is from 1 to 12 replicates The software automatically performs a check to ensure that the number of replicates multiplied by the number of samples does not exceed the number of undefined wells remaining on the plate Sample Location If mapping is manual and samples are defined the locations for each sample replicate must be specified ee eee AI eras aie rre Defining the locations of the samples e Use the NUMERIC and ALPHA keys to enter the well location for Rep 1 of Sample Group 1 Press ENTER to advance to the next replicate or sample group 3 28 Operation Define FORMULA The PowerWaveX supports four types of formulas Cutoff Transformation Validation and General as well as the ability to program variables for use within formulas Up to three types of Validation formulas may be defined Blank Control and Assay Validation To define formulas start at the Main Menu and select DEFINE select the assay press ENTER then select FORMULA Selecting the Formula option on the DEFINE screen Note Formulas created using Bio Tek s Extensions Define Reader Protocol software cannot be edited via the reader front panel CALCULATION STRUCTURE During data reduction formulas are processed in the order shown below The number of permitted formulas
60. T STANDARD OUTLIERS was set to MANUAL in the assay definition the option to edit outliers is presented Choosing whether or not to edit standard outliers e Select NO to include all standards in the curve fit calculations e Select YES to indicate that one or more standard replicates or groups should be temporarily excluded from curve fit calculations gt IfAVERAGE STANDARDS was set to NO in the assay definition one or more standard replicates can be chosen for exclusion PowerWaveX Operator s Manual 3 53 3 54 Choosing to remove replicate 1 of Standard 1 from curve fit calculations Select YES to exclude the replicate from curve fit calculations Select NO to retain the replicate Press ENTER to advance to the next replicate gt IfAVERAGE STANDARDS was set to YES in the assay definition one or more standard groups can be chosen for exclusion Choosing to remove all replicates of Standard 1 from curve fit calculations Select YES to exclude the group from curve fit calculations Select NO to retain the group Press ENTER to advance to the next group Note Each curve fit type requires a minimum number of standards for curve generation 4 for 2 P 4 P cubic and cubic spline 3 for quadratic and 2 for linear and point to point Exercise caution when editing outliers If the assay is left with insufficient standards the curve fit will f
61. TD1 STD1 SMP2 SMP2 B STD2 STD2 SMP3 SMP3 C STD3 STD3 SMP4 SMP4 D STD4 STD4 SMP5 SMP5 E STD5 STD5 F PC PC G NC NC H SMP1 SMP1 Map Direction ACROSS Rep Direction DOWN 1 2 3 4 5 6 7 8 9 10 A STD STD2 STD3 STD4 STDS PC NC SMP1 SMP2 SMP3 B STD1 STD2 STD3 STD4 STD5 PC NC SMP1 SMP2 SMP3 TOmTMUOO Figure 3 4 Examples of Mapping Directions 3 20 11 12 11 12 PC PC SMP5 SMP5 11 12 11 12 SMP4 SMP5 SMP4 SMP5 Operation Start Mapping at Well Location Enter the location of the well that will be the starting point for automatic mapping Entering the starting well for automatic plate mapping e Use the NUMERIC and ALPHA keys to enter a letter or number at the cursor location The valid entry range is from A01 to H12 depending on the number of blanks standards controls and or samples defined in the assay Blank Map There are several blanking methods to choose from Selecting a blanking method e ATR performs an initial reading on air just prior to the plate read and uses that reading as the blank value This value is subtracted from each well on the plate e FULL enables a single blank well or an average of blank wells up to 48 to be subtracted from each well on the plate e CONST Constant allows entry of a user specified absorbance value This value will be subtracted from each well on the plate PowerWaveX Operator s Manual 3 21 e ROW enables a single blank well or an aver
62. THOUT THE EXPRESS WRITTEN PERMISSION OF AN OFFICER OF BIO TEK INSTRUMENTS INC THIS WARRANTY IS VOID UNLESS THE PURCHASE REGISTRATION CARD HAS BEEN COMPLETED AND MAILED TO US WITHIN TEN 10 DAYS OF PURCHASE PowerWaveX Operator s Manual xiii xiv Preface Table of Contents A sed E E E iii NOtICOS 00 a en Heed ee ee ee nee iii COpYPiQht ienee ae e a AEE aae ea aa eaa aa d aaae ds dees E Ean iii TAS E E EE T T A E ETEN iii Restrictions and Liabilities eee cece eeeeeeeeeeeaeeeeeeeeeeesaeeeseeaeeeeeeeeeeesneeeeenenaes iv REVISION Hist Y viso ada darias iv Document Conventions isinsin ii diia iv Warnings and Precautions is cccscce scusctenscceceeescheencectecsugecduceseedcecseinochesteeesesduteetecobee eedieeed v Hazard WamMingS ai iii edi eerde Eida dadida k iTi v PRECAUTIONS AEE TE E TE T TE v Electromagnetic Interference and Susceptibility ooooooconnnnnnnicnnnnnnonncococnnnnnnanancnccncnnnnns vii CE Mark Infortriation assioni penan iynin toro viii Safety SYMDO Saresti cnc a en tati pheasant gushed Ae aar eaaa aena ix Intended USE Statement iii taco xi SPeEcimen Preparate xi Quality Control a a idas xi About this Malal A A es ee xii Registration Card cis narenn nim nese eee eee ead xii AETA I AEE E T E A E A E E E E EE E A EA xiii List of Tables and FOUA S a a ii xxi Chapter 1 Introduction cociendo a duster ca dond 1 1 Introducing the PowerWaveX cccccccececcecneceeeeeeeceeeaeeeeeeeseeeaaeaee
63. WaveX can run computer controlled kinetic assays using up to six wavelengths on the same microplate Spectral scan assays may be performed that use the entire wavelength range from 200 nm to 999 nm at 1 nm increments When using computer control the blanking and data reduction capabilities on the PowerWaveX are suppressed and all data is returned to the user for evaluation Readings higher than 3 000 OD may be transmitted This appendix details the protocols necessary for communicating with the PowerWaveX reader In addition instructions are provided for controlling the reader with Bio Tek s KCjunior or KC4 software packages e Controlling the Reader with KC4 page B 2 e Controlling the Reader with KCjunior page B 4 e Controlling the Reader using Serial Protocol page B 6 e Using the Stop Key to Halt Plate Scans and Reads page B 20 e Status String Format page B 21 Controlling the Reader with KC4 The PowerWaveX can be operated using a computer running Bio Tek s KC4 software Follow the steps below 1 Power off the computer and the reader Connect the appropriate serial cable PN 75053 between the two machines See Table B 1 on the following page for a pin out description of the cable Power up both machines Install KC4 on the computer s hard drive Once installed start KC4 Select System Readers Scroll through the list of Available Readers and select the appropriate PowerWaveX reader model Click the
64. ace Remove the reader from the plastic bag 4 Place all shipping material back into the shipping box for reuse if the instrument needs to be shipped again 5 Inspect the packaging and instrument for shipping damage such as visible dents or scratches 6 Ifthe reader is damaged notify the carrier and your manufacturer s representative 7 Contact Bio Tek s Technical Assistance Center for an RMA Return Materials Authorization number before returning equipment for service Mark the RMA number on the outside of the shipping container See page 1 8 for contact information Top foam 26 x32 y 2 MI Poly Bag Bottom foam Y Figure 2 1 Unpacking and repackaging the reader 2 2 Installation Removal of the Carrier Retention Screw in PowerWaveX Select Models Note The PowerWaveX Select model is shipped with a carrier retention screw that must be removed before the reader is used An allen wrench PN 49329 is supplied for this task See Figure 2 2 below Lift up the front door Using the allen wrench provided remove the carrier retention screw wave washer and flat washer Save all of the shipping hardware Important Replace the carrier retention screw and washers prior to shipment or you will void the manufacturer s warranty Carrier retention screw PN 19953 Wave washer PN 17023 Flat washer PN 17056 Figure 2 2 Removal of the carrier retention screw and accompanying washers from the Po
65. ad is significantly different from the same reading taken during the power up self check The last digit indicates the wavelength and the second to last digit indicates the channel PROBABLE CAUSE e The measurement electronics background noise has changed since the last power up self check This could be caused by a large increase in external ambient light since power up See Probable Cause for Error 0800 e Running the System Test will reset the value to the new baseline Displayed Error Potential Cause ERROR 0A00 Read time air blank out of range Error A00 indicates that the blank full signal reading taken during the current read has changed significantly from the same reading taken during the power up self check PROBABLE CAUSE e The measurement electronics full signal level has changed since the power up self check was last run The flash lamp could be near failure e Running the System Test will restore to new baseline Displayed Error Potential Cause ERROR 0B00 Invalid assay This error indicates that an assay number that is not programmed was selected PowerWaveX Operator s Manual C 5 Displayed Error Potential Cause ERROR 0C00 Printer timeout Error 0C00 indicates that the printer in use is not responding PROBABLE CAUSE e Printer not connected or powered up e Printer s parallel port may not be correctly selected in the on board software See Printing and Data Communications in Chap
66. age of blank wells to be selected for each row The maximum number of blanks is 48 Use manual mapping to position blanks standards controls and samples e COLUMN enables a single blank well or an average of blank wells to be selected for each column The blank OD or average OD will be subtracted from other wells in the column Use manual mapping to position blanks standards controls and samples e P ACROSS enables a blank in every even numbered column to be subtracted from the well to the left of it in every odd column Use manual mapping to set up the appropriate map by placing the standards controls and samples in only the odd columns e P DOWN enables a blank in the B D F and H rows to be subtracted from the well above in the A C E and G rows Use manual mapping to set up the appropriate map by placing the standards controls and samples in rows A C E and G Blanking Constant If the CONST blanking method is selected a Blanking Constant must be defined This value will be subtracted from each well on the plate Entering the Blanking Constant value e Use the NUMERIC keys to enter the value The valid entry range is 0 000 to 3 000 OD Number of Blanks If the FULL COLUMN or ROW blanking method is selected enter the number of blank wells on the plate Entering the number of blank wells on the plate e Use the NUMERIC keys to enter the number of blank wells The valid entry range is 0 to 48 wells
67. ail Printing Results After the assay is selected and standard outliers are edited if necessary the results report can be printed Print Results screen e Ensure that the printer is connected turned on and full of paper e Press YES to print reports or NO to return to the Main Menu Operation Map Report The Map Report contains a matrix in Row x Column format showing the location of every well identifier defined in the plate map EERPEO Bee eels ee Ea Choosing an assay for the Map Report e Press OPTIONS to cycle through the list of available assays or enter the number of the desired assay e Press ENTER to print the report Assay Report The Assay Report lists the assay definition parameters and their current settings Selecting an assay for the Assay Report e Press OPTIONS to cycle through the list of available assays or enter the number of the desired assay e Press ENTER to print the report List Report The List Report lists the all of the assays name and number currently programmed on the reader e Select REPORT from the Main Menu then select LIST to print the report PowerWaveX Operator s Manual 3 55 3 56 Operation Chapter 4 Performance Verification Qualification Tests This chapter discusses the tasks and procedures necessary for verifying and qualifying instrument performance on an ongoing basis A convenient Recommended Test Schedule arranges task
68. ard curve if the assay generates one or NO to suppress this report e Press ENTER to return to the Select Utility Option menu 2 18 Installation SELECTING READ OPTIONS To specify various read time options e From the Main Menu select UTIL gt READ The Prompt for Plate ID screen will appear Press ENTER to cycle through the prompt screens Selecting Read Preferences e If selected at read time PLATE ID prompts for microplate identification SAMPLE ID prompts for an identification for each sample group SAMPLE COUNT prompts for the number of samples on the plate e IfRAPID MODE is disabled the plate reads at a speed designed to ensure low CVs and precise results If Rapid Mode is enabled the plate reads more quickly but may possibly result in higher CVs If an assay favors speed over precision consider reading in Rapid Mode e Pressing ENTER after each selection advances the display e When selections are completed the display returns to the Select Utility Option menu PowerWaveX Operator s Manual 2 19 2 20 Installation Chapter 3 Operation This chapter includes instructions for operating the PowerWaveX and its software Overview The PowerWaveX features a 25 key front panel with a 2 X 24 character LCD Liquid Crystal Display allowing access to the reader s program menus Test results can be sent to an attached printer and or through the reader s bi directional serial
69. ards does not exceed the number of wells on the plate Average Standards If the Number of Standard Replicates setting is greater than 01 the Average Standards screen is presented Choosing whether or not to Average Standards e Select YES to average the replicates for each standard group and then use the group averages when calculating the standard curve e Select No to use the individual standard replicates when calculating the standard curve 3 24 Operation Standard Concentrations Use the Standard Concentration screen to enter the predicted concentration value for each standard group If manual map generation is selected the replicate locations must also be defined eee vee iN ee elas eer eee eee UE Eee Pee els Entering the predicted standard concentrations and well locations if manual map e Use the NUMERIC ALPHA and DECIMAL POINT keys to enter the standard concentration values The valid entry range is 00001 to 999999 The entry cannot exceed 6 characters including the decimal point e If automatic mapping is selected each replicate s location is available for viewing only Pressing ENTER advances to the concentration value entry for the next standard e If manual mapping is selected the location must be defined Pressing ENTER from the standard concentration entry moves the cursor to the Location field Pressing ENTER from the Location field advances to the concentration value entry for
70. ash configuration time out Task control block error PowerWaveX Operator s Manual C 9 C 10 Error Codes Appendix D Sample Reports This appendix contains samples of the different types of reports available on the PowerWavex Reports are automatically generated after a plate has been read see Specifying Data Output and Reporting Options in Chapter 2 for information on selecting report formats In addition Map Assay and Assay List reports can be printed via the REPORT option from the Main Menu This Appendix contains samples of the different types of reports available on the PowerWaveX D 2 Bio Tek Instruments Assay ASSAY1 Date 07 01 99 Lot Time 10 36 10AM Operator Wavelength 405 Temp 30 C Plate 1D 123 COMMENTS Some concentrations may lie outside curve asymptotes x x lt gt around RSLT indicate extrapolated concentration 1 2 3 4 5 6 1 8 9 10 11 12 EEE K rh_ _ c c_ _QOo uOc c c c_ _u u e O CALL POS EQUIV NEG NEG POS NEG NEG POS NEG EQUIV CalcOD 0 031 1 089 1 332 1 057 0 779 0 509 1 320 0 788 0 786 1 389 0 802 1 015 Well BLK STD4 SHPI SMPS SMP9 SMP13 SMP17 SMP21 SMP25 SMP29 SMP33 SMP37 RSLT j lt 24 629 gt 39 786 i 47 526 38 776 29 703 19 394 47 118 29 987 29 947 49 405 30 476 37 439 Bm 0 _ _ _ _ _ _ gt _ _ _______ a e CALL POS EQUIV NEG NEG POS NEG NEG POS NEG EQUIV CalcQD 031 1 092 1 31
71. at the Universal Test Plate cannot The test plate will indicate the absolute amount of light absorbed which will accurately test the linearity of the electronics The liquid test will help detect optical defects such as dirt or contamination that can contribute to errant readings e Ifyou have the Universal Test Plate you will need to run the simple Liquid Test 1 for routine testing e Ifyou do not have a Universal Test Plate test the linearity and repeatability of the reader by preparing a series of solutions of varying absorbances as described in Liquid Test 2 e Ifyou prefer you may use the described dye solution see Table 4 3 The purpose of the recipe is to create a solution that absorbs light in a well defined manner and yields 2 000 OD at full strength when dispensed at 200 microliters in a microplate well e Alternatively any solution that gives a stable color will suffice This includes substrates incubated with an enzyme preparation and then stopped with an acidic or basic solution Some enzyme substrate combinations are shown below e Ifyou need to test a Universal Test Plate reader s performance at 340 nm run optional Liquid Test 3 see Table 4 5 Table 4 2 Typical Enzyme Substrate Combinations and Stopping Solutions Alkaline Phosphate o nitrophenyl phosphate 3N sodium hydroxide beta Galactosidase o nitrophenyl beta D 1M sodium carbonate galactopyranoside Peroxidase 2 2 Azino di ethylbenzothiazolin
72. ation entry screen e Use the NUMERIC keys to enter the time duration in hours minutes and seconds e The maximum duration time is 80 hours Note The Kinetic Interval Duration combination must result in 2 to 40 total kinetic reads PLATE SHAKING A variety of plate shaking options are available on the PowerWaveX reader Shake Mode Selection screen e For non kinetic reads choose FIRST to enable shaking or NONE to indicate no shaking e For kinetic reads choose FIRST to shake before the first read only EVERY to shake before every read or NONE to indicate no shaking PowerWaveX Operator s Manual 3 13 Shake Time If shaking is enabled a Shake Time must be specified Shake Time entry screen e Use the NUMERIC keys to enter the shake duration Valid ranges are 0 1 hours 0 59 minutes and 0 59 seconds e Fora kinetic read with Shake Before Every Read enabled the CONTINUOUS option is available If selected the shake duration is set automatically according to the kinetic interval Shake Speed For the Power WaveX the shake movement is a repeated 0 021 inch movement from the shake position and back Choose from several different speeds including a Variable option Shake Speed selection screen e Select Low for low speed 17 Hz shaking e Select MEDIUM for medium speed 18 Hz shaking e Select HIGH for high speed 19 Hz shaking e Select VARI variable speed shakin
73. aveX Microplate Scanning Spectrophotometer Operator s Manual For R amp D Use and in vitro Diagnostic Use OCTOBER 2002 COPYRIGHT O 2002 PART NUMBER 7261014 REVISION D BIO TEK INSTRUMENTS INC Preface Notices BIO TEK INSTRUMENTS INC Highland Park Box 998 Winooski Vermont 05404 0998 USA Customer Service and Sales Internet www biotek com Phone 888 451 5171 toll free in the U S 802 655 4040 outside the U S Fax 802 655 7941 E Mail sales biotek com Service TAC Phone 800 242 4685 toll free in the U S 802 655 4740 outside the U S Fax 802 655 3399 E Mail labtac biotek com European Coordination Center Bio Tek Instruments GmbH Kocherwaldstr 34 D 74177 Bad Friedrichshall Germany Phone 49 0 7136 9680 Fax 49 0 7136 968 111 E Mail info biotek de COPYRIGHT Copyright 2002 Bio Tek Instruments Incorporated No part of this publication may be reproduced transmitted transcribed stored in a retrieval system or translated into any language without the written permission of Bio Tek Instruments Incorporated TRADEMARKS Bio Tek is a registered trademark and PowerWave X KC4 and KCjunior are trademarks of Bio Tek Instruments Inc Bio Cell is a trademark of Bio Tek Instruments and Bio Cell is patented under U S patent number 5 963 318 Microsoft is a registered trademark and Windows Windows 95 98 2000 NT and XP Word and Excel ar
74. ble Under UTIL 0 cceccecceseesseeseeeeeeeeceeeeeeceseceseceaecsaecaeecaeeeseeneeeaes 2 15 PowerWaveX Front Panel coccion incio eee 3 2 Options Available From the Main Menu cccccesceesseesseesceeseeeseeeeceeeceseenaeenaeeeeenees 3 4 Options Available Under DEFINE 0 cccccccesccesecseeeseeeeeeeeceeseeeeceseenseenseenseceaeeaenaees 3 5 Examples of Mapping Directions c cccsccscessseesseecceesceeeceecesecnsecnseceeecneeeeeeneeaes 3 20 Options Available Under READ cccccceceecsesseesseeseeeseeeeeeeeensecesecesecsaeceaeeeeeeeeneeaes 3 47 Sample Output for the System Test ccccccccsseessessceesceeseeeecseecesecesecnaecaecesecaeeaeeenes 4 5 Sample Universal Test Plate Data Sheet ccccscecssesseeesceeeeeeeceseeeseestecseeeseeeneeseeenes 4 8 Sample Printout Showing the Calibration Plate Analysis cc cesceseeeseeteeeteetees 4 11 Sample Data Sheet Showing Wavelength of Peak in the Interval Between 580 and A aioli eee eae cei E aia eae eee 4 13 Location of Bracket Motor Shaft Mylar Sheet and Main PCB Motor Assembly 4 27 Roller Surface and Hook In Bracket ceseeeseccseesececeeesececeesesecesseeseeeaeeeens 4 28 Matrix Report Showing Well Ids Calculated OD Values and Concentrations Calls on Samples and Comments cccsccssecsseesseeseeseeeseeeeeceeeeeceeeeseenseeneeeaeenseenaes D 2 Curve Fit Report Showing the Curve Method Equation Coefficients and Res quiared Vi ss D 3
75. bration none to all waves selected wavelength 3 200 999 wave terminator 1 lt CR gt calibration terminator 1 lt AZ gt for each wavelength specified for first pass scan none or all waves selected wavelength 3 200 999 wave terminator 1 lt CR gt first pass terminator 1 lt AZ gt data start code 1 lt CR gt for each wavelength specified for scan wavelength 3 200 999 for each well selected in strip 1 or 8 wells comma separator 1 sign 1 or data 4 1934 well strip terminator 2 lt CR gt lt LF gt data terminator 1 lt AZ gt checksum 1 0 255 not ASCII status string 5 only if error The reader sequentially scans the well or strip from the start wavelength through the stop wavelength in increments dictated by the wavelength step At least two wavelengths must be scanned The start must be less than the stop and the step must be less than or equal to the difference PowerWaveX Operator s Manual B 15 B 16 If the calibrate option is selected 1 all wavelengths indicated for the current spectral scan will be calibrated If calibration is not selected any wavelengths selected but not scanned since the instrument was powered up will be calibrated anyway In addition any selected wavelengths with previous errors detected will also be calibrated If no calibration is performed at all the Z gt terminator is ret
76. checksum of the current configuration data This means that the configuration data has changed and the checksum stored is no longer valid The error is produced when outdated versions old of Extensions or Define Assay are used to create an assay configuration file This file is incompatible with the operation code within the instrument s memory The fix for this problem is to recreate the assay definition on the correct version of assay definition software and re download it This can also be caused by a bad Flash memory Reloading the basecode and assay configuration may fix this Displayed Error Potential Cause ERROR 1200 Calibration data missing This error means that AUTOCAL has not been performed after a memory erase or in the case of a new unprogrammed board immediately after the basecode or assay definition download The system must have the AUTOCAL sequence performed This can also indicate a bad Flash memory Displayed Error Potential Cause ERROR 1300 Motor not correctly homed This error will occur if the error 0200 or error 0300 is ignored The situation needs to be fixed before the instrument is used PowerWaveX Operator s Manual C 7 Displayed Error Potential Cause ERROR 1400 Assay incubation error Assay requires incubation but incubation is not available Displayed Error Potential Cause ERROR 1500 Assay incubation error This error indicates that incubator failed to hold temperature within to
77. computer and the reader Connect the serial cable PN 75053 between the two machines See Table B 1 Power up both machines Install KCjunior on the computer s hard drive Once installed start KCjunior Select Setup then Reader1 To select the PowerWaveX reader and define the communications parameters choose the following setup parameters e Reader PowerWaveX e Com Port COM1 or COMA the serial port used for the RS 232 cable connection e Baud Rate 9600 e Data Bits 8 e Parity None e Stop Bits 2 e EOT Character Keep the default number Click the Test Communications button to attempt to establish communications with the reader using the currently defined communication parameters If a Serial Write Error dialog box is displayed an incorrect Com Port may have been selected Select a different port and then repeat this step If the test passes click OK to save the settings and close the dialog box If the test fails follow the directions provided by KCjunior then click Test Communications again To learn more about KCjunior and how to define assays and read plates refer to the KCjunior User s Manual Problems If KCjunior fails to communicate with the reader and displays a serial communications error check the cable plug in location to make sure it matches the setup choices and is not a Null cable If this is suspected add another Null and try again B 4 Computer Control Getting Started w
78. cription e DSR 19S Ne 8 bep ft Ne 9 NCCT NE AE AE l Installation SETTING UP COMMUNICATION PARAMETERS Before serial communication can be initiated between the PowerWaveX and another device such as a host PC running Bio Tek s KC4 or KCjunior software packages the communication parameters must match on both devices The reader s default communication parameters are e Baud Rate 9600 e DataBits 8 e Parity None e Stop Bits 2 The reader s Baud Rate can be changed to 1200 or 2400 if necessary The reader s Data Bits Parity and Stop Bits settings cannot be changed To change the Baud Rate e At the Main Menu select UTIL ESB erre rare Main Menu screen e Atthe Select Utility Option menu select SETUP Selecting Setup e At the Edit Setup Information screen select MORE to see more options including RS232 e Select RS232 to advance to the Select Baud Rate screen PowerWaveX Operator s Manual 2 9 2 10 The top line of the display shows the currently selected Baud Rate Selecting the Baud Rate To change the Baud Rate press the soft key corresponding to the desired setting The top line of the display automatically updates to reflect the new setting To view the reader s other communication settings select VI EW EW Aee eee ee MER Aerea Viewing Other Communication Settings Installation Components LAMP The lamp life
79. curacy is a measure of the absorbance optical density of Calibration Plate wells C01 D04 E02 F05 G03 and H06 with known standard values contained in the Specification Sheet that accompanies each Calibration Test Plate e Linearity Ifthe accuracy specifications are met then the reader also proves to be linear Linearity can also be proven by performing a regression analysis on the OD values in a program such as Microsoft Excel See the instruction on the following page PowerWaveX Operator s Manual 4 11 4 12 In Microsoft Excel open a spreadsheet and label one column Assigned and the next column Observed 1 Enter the Assigned OD data for each glass filter in the first column from the data sheet provided with the Test Plate Analyze one wavelength at a time Next enter the Observed OD values for the same glass filters in the adjacent column Under Tools select Data Analysis and then Regression Use the Regression Input box to enter the Assigned values as the Input Y Range and the Observed OD as the Input X Range Click the OK box and the Summary Output sheet will be displayed An R Square value of at least 0 99 is expected and the results will often be 0 999 or better Repeatability Repeatability is a measure of the instrument s ability to read the same well with minimum variation between two reads with the well in the same location Turnaround Second Repeatability Resu
80. de number e ERROR CODES Error codes indicate the following 0 no fault or error 8 instrument failure perform self test 9 error in assay scan or table definition A error in strip range selection B incubator setpoint error C incubator temperature error E barcode error gt S3 Always ASCII zero 0 gt ETX End Of Text marks the end of the status block Instrument failures are usually accompanied by an error code on the reader display ELx Status Mode Format This status string consists of 5 successive ASCII characters a four byte string representing a hexadecimal status code and then ETX Items described in angle brackets lt gt are indicated by an ASCII digit replacing the last 0 character in the status code Items described in curly braces are indicated by an ASCH digit replacing the next to last 0 character in the status code Fatal errors indicate a hardware failure also shown on the instrument display screen and require recycling of instrument power Note Errors listed on the following pages are common to all reader instruments and may not all be applicable to any single given reader PowerWaveX Operator s Manual B 21 B 22 Fatal Errors TCB NOT AVAIL ERR READ NOT AVAIL ERR NOT AVAIL ERR CHECKSUM ERR DR ALLOC ERR DFLASH TIMEOUT ERR DFLASH ERR CFLASH TIMEOUT ERR HEAP CORRUPTION ERR ATOD ERR Non Fatal Errors NO
81. definito in IEC 417 Caution refer to accompanying documents Attention voir documents d accompanement Achtung siehe Begleitpapiere Atenci n vease los documentos incluidos Attenzione consultare la doc annessa Preface Intended Use Statement The PowerWaveX is an eight channel automated benchtop general purpose Microplate Spectrophotometer that performs in vitro diagnostic analyses of a variety of samples The Performance Characteristics of the data reduction software have not been established with any laboratory diagnostic assay The user must evaluate this software in conjunction with the specific laboratory diagnostic assay This evaluation must include the confirmation that new performance characteristics for the specific assay are met This system is designed for use with a variety of microplate based assays A versatile curve fitting and statistical software program is preloaded on every PowerWaveX plate templates and formulas are automatically combined with the protocol assay setup Data results may be printed out or sent to a computer running a Bio Tek software package such as KCjunior or KC4 for Windows The on board software provides e An easy to use menu driven interface e Endpoint curvilinear regressional and statistical calculations e Curve fitting with 4 parameter cubic quadratic linear cubic spline point to point and 2 P Logit methods e Formula calculations for more complex mathematical
82. diting If set to YES the standard groups are available for editing Operation gt After the assay is run and reports are generated press REPORT from the Main Menu Press RESULT select the assay then press ENTER The EDIT STD OUTLIERS YES NO prompt will appear See Editing Standard Outliers on page 3 53 for further instructions X Y AXIS TYPE After the curve fit type is selected select the X Y Axis Type ee ee FB E EPG ee eee Selecting the X Y Axis Type e Choose the method by which the X and Y axes will be scaled e This option is not available for the 2 P and 4 P curve fit types The X Y scaling for these curves is always LIN LIN EXTRAPOLATION OF UNKNOWNS The PowerWaveX provides the option to extrapolate the curve to evaluate samples outside of the absorbance range defined by the standards Choosing whether or not to extrapolate unknowns e Select YES to enable Extrapolation otherwise select NO e On printed reports extrapolated concentrations RSLT values are surrounded by lt gt for example lt 44 425 gt Note If extrapolation is chosen for the Point to Point curve fit unknown concentrations will be extrapolated linearly from the nearest segment of the curve If the plot includes both increasing and decreasing segments the curve printout will be labeled Ambiguous The resulting values which actually are extrapolated may not be indicated as
83. dition Equal to Subtraction Greater than Multiplication Greater than or equal to OTHER The following additional symbols can be used in formulas Left parenthesis Right parenthesis Logical AND Logical OR MAP The available MAP options depend on the formula type and the current plate map MAP options resemble BLK x mean of the blank wells NC 1 the first NC well or OD every well FUNCTION The following functions can be used in formulas 3 32 Operation VALIDATION FORMULAS Validation formulas can be used to determine whether or not blanks and or controls are valid In addition Assay Validation formulas can be used to determine whether or not the entire assay should be considered valid See Formula Type on page 3 30 for instructions on selecting an assay and accessing the Select Validation Type screen sfe ulefe vlafolr olajrizolw reee cjojw eirjolo alssjaly ojo jajuje Selecting a Validation formula type Control and Blank Validation Formulas Blank Validation is used to ensure that the OD values for the blank replicates or for the blank mean meet certain criteria Control Validation serves the same purpose as Blank Validation but apply to the control replicates or control mean If the criteria are not met results are considered suspect and the message RESULTS INVALID Blank or Control validation failed appears on results reports e One blank val
84. dure bici 4 15 Procedure a ii 4 16 ElqUId Testa a 4 17 EiquId Test 2 e et ae Seg 4 19 Lingarity LeSt A cia sa Seba ond deep in as 4 20 Repeatability Test Borei ntm iei ea e eaten 4 20 Channel to Channel Variation and Alignment Test C e 4 21 Eto UI IA EE Ee Lata a rt aaa acota id 4 22 Procedure Ai a 4 23 Procedure Binig e a e 4 23 Liquid Test 3 Procedure aci a a a 4 23 PowerWaveX Operator s Manual xix XX Repeatability Test A comio ii e eave tiene 4 24 Linearity TestBiiaan a Ai 4 24 Maintenance of Robotic UnitS ooonooccccnnniconoccccnnnoccnononannnn non nc nano cnn rnnnnnnn nan rr nnnr rre 4 25 Appendix A Decontamination and Cleaning ccccccccccccccccccncnenenennnos A 1 Decontamination Procedure cococcconoccccconoccnonononcnanonnnn nono cnn nn n cnn nano n nn rn nn nn rn nn rre A 1 PUTPpOSsS uta ms ie o e o o aS Seas tt A 1 General Considerations imc an A 1 Procedures iii a denied ed A 2 Cleaning Procedure ii a lr A 2 Appendix B Computer Control cocccccccccccccccccncconcccnncnonenonenonenonenenennnos B 1 A A NON B 1 Controlling the Reader with KC4 o oocoocccccccococococococcnocononnonononnncnnnnnnnnnnnnnnnnnnnnnn nn nesteen nannte B 2 Problems A B 2 Getting Started with KA cuotas B 3 Controlling the Reader with KCjUNIOM 0 02 eee eeeeeeeeneeeeeeeeeeeesnaeeeeeeeaeeesneeeeeeaeeeenenaees B 4 ProbleM oia tan B 4 Getting Started with KCjUNIO ooooococcccconococonononanononcncnnnannnn n
85. dures 4 2 Tests outlined in this section may be utilized to confirm initial and ongoing performance of the PowerWavex Your PowerWaveX reader was fully tested at Bio Tek prior to shipment and should operate properly upon initial setup If it is suspected that problems may have occurred during shipment if you reshipped the device or if regulatory requirements dictate that Performance Qualification Testing is necessary the following tests should be performed After the initial confirmation of operation the Universal Plate Test and Liquid Testing should be performed at least monthly The System Self Test and Universal Plate Test can be run from KCjunior via Utilities Diagnostics and the Universal Plate Test can be run from KC4 via System Diagnostics e System Self Test Verifies proper gains bulb operation low electronic noise and optional incubator functionality Test results are sent to an attached printer e Universal Plate Test Confirms the optical accuracy linearity alignment channel to channel variation and wavelength accuracy of the instrument e Liquid Testing Quantifies the channel to channel variation of the instrument using liquids which verifies operation in a way that glass test filters cannot Performance Verification Qualification Tests Routine Procedure To ensure proper operation of the PowerWaveX on an ongoing basis the System Self Test and the Universal Plate Test should be conducted monthly e Se
86. e citrate phosphate buffer pH 2 8 sulfonic acid ABTS Peroxidase o phenylenediamine 0 03N sulfuric acid 4 14 Performance Verification Qualification Tests STOCK SOLUTION FORMULATION The stock solution for Liquid Tests No 1 and No 2 may be formulated from the chemicals listed below or by diluting a dye solution available from Bio Tek See Procedure A or B outlined below for details Procedure A Required Materials e Deionized water e FD amp C Yellow No 5 dye powder typically 90 pure e Tween 20 Polyoxyethylene sorbitan monolaurate a wetting agent e Analytical balance e 1 liter volumetric flask Table 4 3 Stock Solution Formulation for Liquid Test Nos 1 and 2 FD amp C Yellow No 5 powder 0 092 g DI Water to bring volume to 1000 ml Preparation of Stock Solution 1 Weigh out 0 092 gram of FD amp C No 5 yellow dye powder into a weigh boat 2 Rinse the contents into a 1 liter volumetric flask 3 Add 0 5 ml of Tween 20 4 Make up to 1 liter with DI water cap and shake well gt This should create a solution with an absorbance of about 2 000 when using 200 ul in a flat bottom microwell The OD value result will be proportional to the volume in the well and the amount of FD amp C No 5 dye used You can use a larger or smaller well volume or add more dye or water to adjust the solution Note that too small a well volume may result in increased pipetting related errors PowerWaveX Operator s
87. e incubator heaters off PowerWaveX Operator s Manual B 17 GET TEMPERATURE J This command returns the current temperature in the incubation chamber host valid limits response no arguments lt ACK NAK gt temperature 2 status string 5 The reader returns the current temperature as a 2 byte integer value not ASCII so low byte first Temperature is returned scaled up by 10 i e 370 indicates a temperature of 37 0 degrees Celsius If the instrument does not have incubation or an incubation error has been detected 0x0000 will be returned as temperature data An incubator temperature error will then be flagged and returned with the standard status response string Otherwise the current temperature averaged over four thermal zones is returned as defined above The reader will accept and process the Get Temperature command at any time including during a read cycle The temperature response will NOT interrupt a plate data response stream however If a Get Temperature command is received during a data transmission it will not be processed until the transmission has completed i e after the lt Z gt checksum combination has been sent GET PLATE BARCODE s B 18 This command returns the barcode for the microplate currently in the instrument carrier host valid limits response iS no arguments lt ACK NAK gt barcode 33 status string 5 Computer Control The barcode is returned as a 33
88. e registered trademarks of Microsoft Corporation All other trademarks are the property of their respective holders PowerWaveX Operator s Manual iii RESTRICTIONS AND LIABILITIES Information in this document is subject to change and does not represent a commitment by Bio Tek Instruments Inc Changes made to the information in this document will be incorporated in new editions of the publication No responsibility is assumed by Bio Tek for the use or reliability of software or equipment that is not supplied by Bio Tek or its affiliated dealers REVISION HISTORY 2 99 Incorporated Manual Updates 8 99 Added support for PowerWave Select model Enhanced Chapter 4 Performance Verification amp Qualification Assigned each of the recommended maintenance tasks the Installation Performance and or Operation Qualification classification Changed PowerWave to PowerWaveX throughout Updated Notices Chapters 1 through 4 and Appendices B and C Incorporated manual update for periodic mantenance of robotic units in Ch 4 Added Cleaning section to Appendix A DOCUMENT CONVENTIONS This manual uses the following typographic conventions Example Description A This icon calls attention to important safety notes Warning A Warning indicates the potential for bodily harm and tells you how to avoid the problem Caution A Caution indicates potential damage to the instrument and tells you how to avoid the problem DEF
89. e solutions especially with small volumes Agitate the microplate before reading to help bring this problem within acceptable limits Use Tween 20 if possible or some other wetting agent to normalize the meniscus Some solutions develop menisci over a period of several minutes This effect varies with the brand of microplate and the solution composition As the center of the meniscus drops and shortens the light path the density readings change The meniscus shape will stabilize over time Recommended Test Schedule The schedule shown in Table 4 1 defines the factory recommended intervals for performance testing for a microplate reader used for one shift seven days a week Note The risk factors associated with your tests may require that the Performance and Operation Qualification procedures be performed more or less frequently than shown below Table 4 1 Recommended Test Schedule Performance Performance Operation Qualification Qualification Qualification Monthly Quarterly Annually Installation Qualification System Self Test p 4 4 Universal Plate Test p 4 7 Liquid Test 1 p 4 17 Liquid Test 2 p 4 19 Liquid Test 3 p 4 22 Optional for 340 nm Robotic Lube p 4 25 Every six months or after 10 000 cycles Run Liquid Test 2 only if you do not have a Universal Test Plate If you run Liquid Test 2 or 3 you do not have to also run Liquid Test 1 Installation and Performance Qualification Proce
90. e the greyzone and cutoff values as well as the equations used to assign calls to samples See Specifying Data Output and Reporting Options in Chapter 2 for information on Column Reports The greyzone is a definable area around the cutoff value Samples falling within an area defined by the greyzone ex 5 of the cutoff value could be considered questionable or equivocal EQUIV Defining a greyzone around the cutoff value 3 38 Use the NUMERIC keys to enter the greyzone percentage The valid entry range is from 00 to 99 An entry of 00 indicates no greyzone although a sample equal to the cutoff value will still receive the EQUIV call See POS NEG Calls on the next page for information on how calls are assigned Operation POS NEG Calls After the greyzone is defined calls for the sample wells POSitive N defined EGative EQUT Vocal must be s aulol gt cujrlorej jasaj eels Ee E eva ear alee We et ey Defining the POS NEG EQUIV calls for samples e Select POS or NEG to select the call that will be assigned to samples greater than the cutoff value plus the greyzone e If for example POS is selected in the screen shown above calls will be assigned according to the following equations SMP represents the sample wells EQUIV SMP lt CUTOFF CUTOFF GREYZONE AND SMP gt CUTOFF CUTOFF GREYZONE POS SMP gt CUTOFF CUTOFF GREYZON
91. e to perform routine maintenance This Warranty is VOID if the Product has been repaired or altered by persons not authorized by Bio Tek or if the Product has had the serial number altered effaced or removed This Warranty is VOID if any of the Products has not been connected installed or adjusted strictly in accordance with written directions furnished by Bio Tek Batteries fuses light bulbs and other consumable items used in any of the Products are not covered by this Warranty Software utilized in conjunction with any of the Products is not covered by the terms of this Warranty but may be covered under a separate Bio Tek software warranty We will continue to stock parts for a maximum period of five 5 years after the manufacture of any equipment has been discontinued Parts shall include all materials charts instructions diagrams and accessories that were furnished with the standard models THIS WARRANTY CONTAINS THE ENTIRE OBLIGATION OF BIO TEK INSTRUMENTS INC AND NO OTHER WARRANTIES EXPRESSED IMPLIED OR STATUTORY ARE GIVEN PURCHASER AGREES TO ASSUME ALL LIABILITY FOR ANY DAMAGES AND OR BODILY INJURY OR DEATH THAT MAY RESULT FROM THE USE OR MISUSE OF ANY EQUIPMENT OR INSTRUMENT BY THE PURCHASER HIS EMPLOYEES AGENTS OR CUSTOMERS OTHER THAN THE EXPRESS WARRANTY CONTAINED HEREIN WE SHALL NOT BE RESPONSIBLE FOR ANY DIRECT OR CONSEQUENTIAL DAMAGES OF ANY KIND THIS WARRANTY SHALL NOT BE CHANGED OR MODIFIED IN ANY WAY WI
92. eeeseseeaeaeeeeeeeseeseaeeeeess 1 1 Variations toria dlrs 1 1 Hardware Features cutis tino 1 2 Software Features iia ora od 1 2 Package Content cion Dri 1 3 Optional ACCESSOMES cito A a at 1 3 SPECINCALIONS EAE E EE doce ccdac svat cons oudedscat se E E EE 1 4 TECHNICAL SUPPOMbesss etched este he ee aes vase tia ee eta ee eens 1 8 Chapter 2 Installation ii 2 1 Operating Environment ccccccccesssceceeeeeeeeneeeeeeeeeeeceaeaeeeeesesecaeaeeeeeeesecenaeeeeeeeseeeesaees 2 1 Unpacking and Repackaging the Instrument c cccceeeeeeeeeeece cece eeseeeaeeeeeeeeeeeenteaeees 2 1 PowerWaveX Operator s Manual XV Removal of the Carrier Retention Screw in PowerWaveX Select Models 2 3 After Unpacking Verify Performance cccecceceeeeeeeeneeceeeeeeeeeeeaeeeeeeeseseesueeeeeeeees 2 4 Before Repackaging the InStruMent ooooccccnnnnnnnnocccccnccnocononononnncnnnnnanononnncnnnnnnnn cnn 2 4 Adjusting the Reader s Wavelength Table SettingS oooonooocccnnnnnnnicoccccnnnocanananccnncnanannns 2 4 Printing and Data COMMUNICATIONS oooooonccnnnncccnnnoccnononananonono no nano cnn rnnnn nn nn nn rra rn 2 6 Installing a Printer ios asa cs 2 6 Parallel Port Pin Defimition o comiioion ci a iei ae 2 7 Serial Port for Communication with Other Devices oooooococonoccnnnococcnonancnononancnanancnnnnno 2 8 Connecting to the PC and Establishing Communication 2 8 Serial Port Pinout DescriptiOM
93. ent to fill out and mail the postage paid Warranty Registration card By sending in the registration card you will be assured of receiving prompt information on product enhancements xii Preface Warranty This Warranty is limited and applies only to new products except for computer based software which is covered under a separate Warranty Policy manufactured by Bio Tek Instruments Inc Bio Tek Bio Tek makes no warranty whatsoever regarding the condition of used products Bio Tek warrants the instrument hereinafter collectively referred to as Products or Product for a period of one 1 year from the original purchase date against defective materials or workmanship This Warranty is limited to the original purchaser the Purchaser and cannot be assigned or transferred All claims under this Limited Warranty must be made in writing to Bio Tek Attention Service Department Purchaser must ship the Product to Bio Tek postage pre paid Bio Tek shall either repair or replace with new or like new at its option and without cost to the Purchaser any Product which in Bio Tek s sole judgment is defective by reason of defects in the materials or workmanship This Warranty is VOID if the Product has been damaged by accident or misuse or has been damaged by abuse or negligence in the operation or maintenance of the Product including without limitation unsafe operation operation by untrained personnel and failur
94. erWaveX Operator s Manual lification Tests Appendix A Decontamination and Cleaning This appendix contains the procedures for decontaminating and cleaning the PowerWaveX Decontamination Procedure If the PowerWaveX is to be shipped after being exposed to potentially hazardous material it should be decontaminated The following procedure outlines how to decontaminate the instrument before packaging and shipment PURPOSE Decontamination minimizes the risk to all who come in contact with the reader during shipping handling and servicing It is also required by the U S Department of Transportation regulations GENERAL CONSIDERATIONS e Any laboratory instrument that has been used for clinical analysis is considered a biohazard and should be decontaminated prior to handling Intact skin is generally considered an effective barrier against infectious organisms however small abrasions and cuts may not be always be visible Prophylactic gloves must be worn when handling instruments that have not been decontaminated Gloved hands should be considered contaminated at all times and must be kept away from eyes mouth and nose at all times e Mucous membranes are considered prime entry routes for infectious agents Wear eye protection and a surgical mask when there is a possibility of aerosols e Fating and drinking while decontaminating instruments is not advisable Important Disconnect the unit from the power sup
95. es that may appear on the reader If an error is displayed and you cannot solve the problem call Bio Tek s Technical Assistance Center Refer to Chapter 1 Technical Support for contact information Error Codes An error code is displayed on the microplate reader as a four digit identifier The first digit is usually 0 or A A 0 denotes a non critical error which means the instrument will still respond to keypad input An A denotes a serious error which requires that the reader be powered down and then powered back up before any diagnostics can be performed Displayed Error Potential Cause ERROR 0200 Plate carrier did not find the home sensor ERROR 0201 Order sorting filter wheel did not find the home sensor ERROR 0202 Robot lid lift did not find the home sensor Y axis could not find the optosensor Errors 0200 0201 0202 and 0204 indicate that an axis was not able to correctly travel to its home position Both axes have optical sensors which when interrupted indicate that the specific axis has been successfully homed PROBABLE CAUSE e Carrier Axis Case defective sensor The X axis movement is limited so that the optical sensor cannot be interrupted e Order Sorting Wheel Case defective sensor The order sorting filter wheel movement is limited so that the sensor cannot be interrupted Note In cases where a sensor is not functioning the motor will drive the axis to its mechanical stop and generate s
96. f desired Press ENTER to continue The Number of Assays entry screen will appear Operation Entering the number of assays to include in the Panel e Specify the number of assays to include in the panel 1 to 8 e Press ENTER to continue The Mapping Direction selection screen will appear Are AAA Perrera pa Choosing the Panel s mapping direction e This option ensures that all assays will be mapped in the same direction Select DOWN or ACROSS Note The original map directions for the pre defined assays are overridden by the Panel s direction If the assays include replicates they will follow the Panel map direction After selecting the mapping direction of the assays choose which assays to include in the panel di A CANTEN ESPA Selecting the assays to include in the panel e Press OPTION to cycle through the assay numbers and names or use the NUMERIC keys to enter an assay number Press ENTER to make a selection e After selecting an assay its starting location must be defined PowerWaveX Operator s Manual 3 45 Defining the starting location for the selected assay e Use the ALPHA and NUMERIC keys to choose the well location to begin the assay Wells A01 through HO1 are valid for ACROSS mapping A01 through A12 are valid for DOWN mapping e This process will be repeated for each assay within the panel Remain aware of the total number of controls standards and blanks that were originally
97. formation on error codes e A daytime phone number e Your name and company information e A fax number and or an email address if available e Ifyou need to return the reader to Bio Tek for service contact Bio Tek for a Return Materials Authorization RMA number and be sure to repack the reader properly see Chapter 2 Installation Phone Support You can telephone the Technical Assistance Center between 8 30 AM and 5 30 PM Eastern Standard Time EST Monday through Friday excluding holidays Bio Tek Instruments Main Number 802 655 4040 Technical Assistance Center 800 242 4685 Written Communication If you prefer you may write a letter with your comments and send it to Bio Tek Instruments Inc Technical Assistance Center Highland Park Box 998 Winooski Vermont 05404 0998 USA Introduction Facsimile Support You may send a fax with your questions or requests for help 24 hours a day to the following numbers Technical Assistance Center 802 655 3399 Electronic Communication Electronic communication is available via the following E Mail labtac biotek com Internet Site www biotek com PowerWaveX Operator s Manual 1 9 Introduction MT Chapter 2 Installation This chapter includes instructions for unpacking and setting up the PowerWaveX instructions for connecting to printers and or serial devices and descriptions of some of the reader s components Operating Environment For opti
98. g 1 second of each speed repeated Operation KINETIC ANALYSIS For kinetic reads there are three calculation options to choose from Kinetic Data Analysis selection screen e Select RATE to apply a linear fit to calculate the maximum slope in mOD min based on the number of kinetic points specified e Select R SOR to calculate the R squared value at the maximum slope based on the linear curve fit and the number of kinetic points specified e Select ONSET to calculate the time it takes for each well to reach the specified onset optical density Kinetic Points For the kinetic analysis selections RATE and R SQR specify the number of sequential kinetic points used to calculate the steepest Rate or the R squared value at the steepest Rate Ble Ale le eer e OIE e Defining the number of Kinetic Points for use in calculations e Select ALL to indicate all of the kinetic reads e To specify a subset of kinetic reads select POINTS then use the NUMERIC keys to input the number of points The valid entry range is 2 to the total number of kinetic reads Note Ifthe number of kinetic reads is changed during assay definition you may need to adjust the Kinetic Points setting The number of Kinetic Points must be less than or equal to the total number of kinetic reads PowerWaveX Operator s Manual 3 15 Onset OD For the Kinetic Analysis selection ONSET an Onset OD is required SEES BASE ERA
99. he PowerWaveX contains several global configurable options such as date and time report output and plate reading preferences These options are accessed via the Select Utility Option menu see below and include e TESTS Runa SYSTEM test to check reader optics or the CALPLATE test to check alignment and calibration Select CHKSUM to perform an internal checksum test and to display the version and configuration information for the currently installed software e SETUP Set the current date and time as well as the date and time formats e OUTPUT Specify where plate data should be sent to a printer a computer or to both Additional options include report format Column and or Matrix and whether or not to print standard curves e READ Enable or disable read time prompting for Plate ID Sample ID and Sample Count Specify whether or not to read in Rapid mode UTI ECT UTILITY OPTION TS SETUP OUTPUT READ PROMPT FOR PLATE ID YES YES NO REPORT OUTPUT PRINT PRINT COMPUTER EDIT SETUP INFORMATION DATE TIME LAMBDA MORE EST CHKSUM CALPLATE Figure 2 5 Options available under UTIL PowerWaveX Operator s Manual 2 15 SETTING THE DATE AND TIME 2 16 To set the current Date and Time and or to change their formats e From the Main Menu select UTIL gt SETUP The Edit Setup Information menu will appear Selecting DATE from the
100. he wells as A1 BLK B1 NC C1 PC D1 SMP1 E1 SMP2 and so on gt ACROSS maps the wells as Al BLK A2 NC A3 PC A4 SMP1 A5 SMP2 and so on e Additional examples of mapping directions are shown in Figure 3 4 3 18 Operation Replication Direction The Replicate Direction is the direction Down or Across in which blank control standard or sample replicates will be mapped on the plate O AAC OCA IL perro rr PRA Selecting the direction for replicate mapping e Select DOWN to map replicates down the plate columns e Select ACROSS to map replicates across the plate rows e For example if the plate contains two blanks BLK two negative controls NC two positive controls PC and samples in duplicate gt DOWN maps the wells as Al B1 BLK C1 D1 NC E1 F1 PC G1 H1 SMP1 gt ACROSS maps the wells as A1 A2 BLK A3 A4 NC A5 A6 PC A7 A8 SMP1 and so on e Additional examples of mapping directions are shown on page 3 21 PowerWaveX Operator s Manual 3 19 Map Direction DOWN Rep Direction DOWN 1 2 3 4 5 6 7 8 9 10 STD1 STDS SMP2 STD1 STDS SMP2 STD2 PC SMP3 STD2 PC SMP3 STD3 NC SMP4 STD3 NC SMP4 STD4 SMP1 SMP5 STD4 SMP1 SMP5 TOTMUOOU gt Map Direction ACROSS Rep Direction ACROSS 1 2 3 4 5 6 7 8 9 10 A STD1 STD STD2 STD2 STD3 STD3 STD4 STD4 STDS STDS NC NC SMP1 SMP1 SMP2 SMP2 SMP3 SMP3 SMP4 SMP4 TOTMUOOU Map Direction DOWN Rep Direction ACROSS 1 2 3 4 5 6 7 8 9 10 A S
101. ibed on page 4 23 This test is optional as the front end of the instrument shows good correspondence at various wavelengths Thus if the device performs properly using the test plate at 405 nm it will also perform adequately at 340 nm Required Materials e Deionized Water e Pipettes e Costar 3590 Flat Bottom Microplate e Beakers and Graduated Cylinder e Analytical balance e Tween 20 Polyoxyethylene sorbitan monolaurate e Phosphate Buffered Saline with Tween 20 PBS Buffer Solution Use Sigma P 3563 packets which will be adequate for one liter of PBS solution each Procedure B or prepare a 10X concentrate per Table 4 5 Procedure A e B NADH Powder B Nicotinamide Adenine Dinucleotide Reduced Form Sigma bulk catalog number N 8129 or pre weighed 10 mg vials Sigma number 340 110 Stock Buffer Solution Formulation Table 4 5 Phosphate Buffered Saline 10X Concentrate Solution Na2HPO4 anhydrous 1 15 grams Add Deionized water to bring to 100 ml 4 22 Performance Verification Qualification Tests Procedure A Mix 45 ml of deionized water with 5 ml of the concentrated PBS solution from Table 4 5 in a beaker Add 10 mg of the B NADH powder and mix thoroughly This is the high level test solution Procedure B Add 50 ml of a PBS solution prepared from the Sigma powder to a beaker Add 10 mg of the B NADH powder and mix thoroughly This is the alternate high level test solution Liquid Te
102. ibration Plate Analysis report in Figure 4 3 Performance Verification Qualification Tests CALIBRATION PLATE ANALYSIS 05 50PM 08 30 02 Read Lambda 405 Operator ID AR SO Notes 132427 Al Top Left Alignment Results A01 0 000 PASS A12 0 001 PASS H01 0 000 PASS H12 0 000 PASS Al Top Left Accuracy Results co1 D04 E02 FO5 G03 H06 STANDARD 0 160 0 617 1 221 1 804 2 210 2 919 DATA 0 155 0 623 1 230 1 818 2 228 2 921 RESULT PASS PASS PASS PASS PASS PASS Repeatability Results col D04 E02 F05 G03 H06 READ 1 0 155 0 623 1 230 1 818 2 228 2 921 READ 2 0 155 0 623 1 230 1 819 2 228 2 923 RESULT PASS PASS PASS PASS PASS PASS H12 Top Left Accuracy Results co1 D04 E02 FO5 G03 HO6 STANDARD 0 160 0 617 1 221 1 804 2 210 2 919 DATA 0 163 0 627 D227 1 807 2 223 2 931 RESULT PASS PASS PASS PASS PASS PASS Repeatability Results col D04 E02 F05 G03 HO6 READ 1 0 163 0 627 1 227 1 807 2 223 2 931 READ 2 0 162 0 627 1 227 1 808 2 224 2 923 RESULT PASS PASS PASS PASS PASS PASS Figure 4 3 Sample printout showing the calibration plate analysis The Calibration Plate Analysis Report contains results for the following e Alignment This portion of the test measures the alignment of the microplate carrier with the optical path A reading gt 0 015 represents an out of alignment condition Wells A01 A12 H01 and H12 are the only valid alignment holes for the reader on the PN 7260522 Calibration Test Plate e Accuracy Ac
103. idation formula can be defined e Up to 4 control validation formulas can be defined e Define the plate map via DEFINE MAP before creating blank or control validation formulas e Blank Control validation can be performed on individual replicates BLK PC or on the group mean BLK x NC x Examples If an assay protocol states that e Each Blank well should have an OD less than 0 050 The formula is BLK lt 0 050 e Each Positive Control replicate must fall within the OD range of 1 000 to 2 500 This can be accomplished with one formula PC gt 1 000ANDPC lt 2 500 or with two separate formulas PC gt 1 000 and PC lt 2 500 e The Negative Control mean must have an OD less than 0 100 The formula is NC x lt 0 100 PowerWaveX Operator s Manual 3 33 Number of Required Blanks Controls When a blank or control validation formula is defined enter the number of blanks or controls that must meet the criteria established by that formula efe felelj foley vasjeio rjejojijrjclajrje s aleje eje Entering the number of blanks controls that must be found valid e Use the NUMERIC keys to enter the number of blanks or controls that are required to be found valid in order for the results to be valid For example if an assay states that two out of three PC wells must be valid enter 02 e The range is 1 through the number of defined replicates of the blank or control Assay Validation Formulas
104. ight levels and electronic noise at all wavelengths fall within factory acceptance criteria and accomplishes this by measuring the air and dark readings and evaluating them to ensure they fall within specified ranges The reader automatically runs an internal System Test each time it is powered on An error will be displayed if the power on System Test fails No report will be produced at the power on System Test To obtain a report of the System Test values for either periodic testing documentation or troubleshooting Figure 4 1 start at the Main Menu and press UTIL Y TESTS Y SYSTEM The PowerWaveX will conduct the instrument s System Test and report results in a pass fail format See Appendix C for a list of possible error codes Photodiodes The Optics portion of the System Self Test confirms that the eight reading and one reference channels have adequate signal range without saturating the electronics Xenon Flash Lamp The Optics test also indicates if the xenon flash lamp is within operational limits Incubation If the PowerWaveX has the 4 Zone incubation option installed the System Self Test will verify the four thermistors and compare these readings to internal voltage references to confirm proper operation Performance Verification Qualification Tests Operator ID SARA Notes Wez 27 02 51PM 08 30 02 SYSTEM SELF TEST Lambda 200 Gain 1 64 Resets 2 Channel Ref 1 2 3 4 Air 31445 37883
105. in Four groups of 96 readings are taken at 100 ms intervals This data is reduced to four averages that cannot vary by 20 counts or a 0700 error will result The outgoing production specification for this test is 12 counts of variation The unit will not flag an error until a 20 count variation is seen PROBABLE CAUSE e External signals getting into the measurement circuit The bottom and top shrouds should be correctly installed e This problem in a correctly assembled unit could indicate a bad main or analog board Failure indicates excessive variation in the dark current background noise levels of the measurement circuit e Defective ground on the track zone 3 ground wire e Malfunctioning incubation board e Defective photodetector on the analog PCB Displayed Error Potential Cause ERROR 0800 Reader failed offset test Error 0800 indicates that the measurement electronics dark current offset is outside of acceptable limits at maximum gain The noise signal level must be between 144 and 2019 counts PROBABLE CAUSE e Ambient light leak track access door open during self check e The analog board could be dirty or defective e The digital board power supplies could be outside their intended values e The photodiode could be defective Error Codes Displayed Error Potential Cause ERROR 0900 Read time dark value out of range Error 0900 indicates that the dark current value taken during the current re
106. in a plate read press the READ key To abort the read press the STOP key Startup Screen The PowerWaveX performs a self test when powered on displaying the Startup screen until initialization is complete During this period no keys are active If the instrument fails to pass the self test a beep will sound and an error code will display Refer to Appendix C to interpret error codes Contact Bio Tek Instrument s Technical Assistance Center for assistance on troubleshooting errors see page 1 8 for contact information PowerWaveX startup screen PowerWaveX Operator s Manual 3 3 Main Menu 3 4 Once the system is initialized the Main Menu is displayed The keypad s four SoftKeys located below the on screen menu options READ DEFINE REPORT and UTIL are activated and may be selected BERE I See ps erase relajo ofejrjx wje falee eleja Main Menu screen Press the SoftKey that corresponds to a displayed menu option to activate that option ECT ASSAY NUMB READ Choose an assay for plate reading Alternatively press the key labeled READ on the keypad DEFINE Define the data reduction parameters for a new assay or modify an existing one REPORT Print a Result Map Assay or List report UTIL Runa System Test or Calibration Plate Test Set up various global configuration options such as date and time report output and plate reading preferences
107. ing instead of a signed numerical value Checksum calculation starts with the first byte AFTER the data start code up through and including the lt Z gt data terminator code The checksum is transmitted as an integer data byte not ASCII If any invalid arguments are sent by the host computer a lt NAK gt response is returned by the reader The Set Geometry command must be sent before any Scan Plate commands if a microplate other than the standard 96 well plate is to be used Computer Control SELF TEST This command causes the reader to perform a system self test and calibration This should be performed any time the wavelength configuration is changed M command host valid limits response ta no arguments lt ACK NAK gt data stream status string 5 The reader responds by sending a variable sized stream of ASCII character data representing various calibration and test results This data stream is followed by the standard status response string SET TEMPERATURE T This command sets the incubation chamber temperature setpoint host valid limits response T lt ACK NAK gt data bytes 1 2 temperature setpoint 0 22 50 status string 5 If the instrument is not equipped with a working incubator or if the indicated temperature setpoint is out of range an incubator setpoint error will be returned with the standard status response string A setpoint of zero will turn th
108. ings are accurate This can be proven by analyzing the data in a program such as Microsoft Excel Alignment of the plate carrier and standard microplates are confirmed by the four corner positional accuracy check Channel to channel variation can be tested by completing the turnaround test This tests the reader s ability to read samples accurately in different plate positions Wavelength setting accuracy To check the accuracy of wavelength settings use the Universal Test Plate The Test Plate provides a multiband test filter Didymium glass V10 in location C6 PowerWaveX Operator s Manual 4 7 Requirements To run the Universal Plate Test from the reader front panel you need Bio Tek s Universal 7 Filter Test Plate PN 7260522 with its accompanying Data Sheet shown in Figure 4 2 below This test plate can be used for testing the reproducibility linearity and alignment of your Bio Tek autoreader The following data has been recorded by a N I S T traceable spectrophotometer WAVELENGTH nm Well 405nm 450nm 490nm 550nm 620nm 630nm 690nm 750nm c1 0 139 0 134 0 130 0 127 0 133 0 134 0 125 0 130 E oso rl aseos elos 0 852 1 256 1 218 O 742 1 628 1 60 pal 2 837 2519 2 445 2 415 2 357 2 324 1 932 1 595 Set 4796 Serial 174819 o O o mo o o a o o o i Ke N gt E Na a ly a amp N hb ob lo N or ja o o 00 alo ob o lun N O
109. inouts Parallel Port Pinout 2 D0 SS ENG 6 D4 wa Jend pa Joe GNnD 9 D7 22 GND a3 ne J S PowerWaveX Operator s Manual 2 7 SERIAL PORT FOR COMMUNICATION WITH OTHER DEVICES The PowerWaveX has a 25 pin serial RS 232 port located on the rear panel of the instrument see Figure 2 3 for an illustration of the serial cable connection The serial port allows the reader to communicate with a computer using standard communications software and or RS 232 protocols Appendix B contains information on required protocols for computer control of the reader CONNECTING TO THE PC AND ESTABLISHING COMMUNICATION 1 Power down the computer and the reader 2 Connect the appropriate serial cable to both machines see Figure 2 3 The serial port on the reader is a DTE configuration with a 25 pin pin male D sub connector The connector s pinout is illustrated in Table 2 2 3 Power up the reader and the computer Each time the reader is turned on it performs a system self test If the self test completes successfully the reader is ready for use If the test fails note any error codes and contact Bio Tek 4 Ensure that the PowerWaveX and the computer are operating with the same communications parameters as described on the following page SERIAL PORT PINOUT DESCRIPTION 2 8 Table 2 2 describes the reader s serial RS 232 pin connection Table 2 2 Serial Pinout Description Serial Pin Des
110. is not installed correctly Do not power up the instrument until the voltage input range to be used is indicated correctly by the peg 2 13 Adjusting the Fuse Configuration or Fuse Replacement Both U S and European fuses are installed in the reader s power input fuse module The reader s fuses are configured at the factory prior to shipping Use the following procedure if you need to change the fuse configuration or replace fuses A failed fuse is usually an indication of another problem which a new fuse is not likely to fix Contact Bio Tek Technical Services if the fuse replacement fails to rectify the problem 1 Use a small flat blade screwdriver to remove the fuse module from the power input module The fuse module has two fuse configurations e The U S configuration has a Hot fused 75 amp slo blo PN 46023 e The European configuration has both Hot and Neutral fused 315 amp slo blo T 5 X 20 mm PN 46051 To replace a defective fuse pop out the old fuse and replace it with the correct new one The configuration of the fuse module U S or European is determined by which fusing network is facing the inside of the power input module refer to Figure 2 4B To change the configuration remove the Phillips head screw that anchors the fuse holder Remove the fuse holder and turn it over Replace the screw and reinstall the fuse module in the power input module Installation Other Utility Options T
111. ith KCjunior The following instructions briefly describe how to read a plate using KCjunior Refer to KCjunior s Help system and User s Guide early and often to learn how to create protocols assign well identifiers read plates print reports and more To read a plate using KCjunior 1 Ze Click Read Plate from KCjunior s main screen The Read Plate Dialog will appear If desired enter a Results ID and a Plate Description and then click Read Plate The Protocol Definition dialog will appear Select a Read Method Type of Endpoint Kinetic Multi Wavelength or Spectrum Define the wavelength s at which the plate will be read Select a Plate Geometry of 8x12 96 well plate or 16x24 384 well plate Define other reading parameters as necessary Click the Help button for assistance When complete click OK to return to the Read Plate dialog If desired enter a Plate ID Place the plate on the carrier then click OK to start the plate read e The plate will be read and then the raw data results will display in KCjunior Print the raw data by selecting Plate Print Results e To analyze or manipulate results a protocol should be defined Refer to KCjunior s Help system or User s Guide for instructions PowerWaveX Operator s Manual B 5 Controlling the Reader Using Serial Protocol At baud rates of 1200 2400 and 9600 the PowerWaveX is capable of sending and receiving data through its serial port RS 232C The ba
112. lanks e Location of Blanks e Number of Standards e Number of Standard Replicates e Averaging of Standards e Concentration and Location of Standards e Number of Controls e Control Type Definition e Number of Control Replicates e Control Location e Number of Samples e Number of Sample Replicates e Sample Location PowerWaveX Operator s Manual 3 17 MAP GENERATION Map Generation represents the method by which blanks controls standards and or samples are assigned to specific locations on the plate Selecting Manual or Automatic map generation e Select AUTO to instruct the software to automatically generate a Plate Map after the blanks controls standards and or samples have been defined e Select MANUAL to indicate that the well assignments will be performed manually by the user at Define and or Run time Note Press Shift Clear at the MAP GENERATION screen to clear a previously defined map Mapping Direction The Mapping Direction is the direction Down or Across in which blank control standard or sample groups will be mapped on the plate COCOS eee eet ae eta eae Selecting the direction for group mapping e Select DOWN to map groups down the plate columns e Select ACROSS to map groups across the plate rows e For example if the plate contains one blank BLK one negative control NC one positive control PC and samples with one replicate gt DOWN maps t
113. ld be below 6 in normal read mode Endpoint or Kinetic with an interval lt 20 seconds Absorbance Measurement Range 0 000 to 4 000 Abs Accuracy 0 000 to 2 000 Abs 1 0 010 Abs 2 000 to 3 000 Abs 3 0 010 Abs Linearity 0 000 to 2 000 Abs 1 2 000 to 3 000 Abs 3 Repeatability 0 000 to 2 000 Abs 1 0 005 Abs 2 000 to 3 000 Abs 3 0 005 Abs Spectral Scanning or Kinetics with short intervals When read intervals from 8 seconds to 20 seconds are selected or spectral scanning is performed the specifications are as follows Absorbance Measurement Range 0 000 to 3 000 Abs Accuracy 0 000 to 2 000 Abs 1 0 010 Abs 2 000 to 2 500 Abs 3 0 010 Abs Linearity 0 000 to 2 000 Abs 1 2 000 to 2 500 Abs 3 Repeatability 0 000 to 2 000 Abs 1 0 005 Abs 2 000 to 2 500 Abs 3 0 005 Abs PowerWaveX Operator s Manual 1 5 1 6 384 Well Plate Reading The following specifications apply to 384 well flat or round bottom plates single wavelength endpoint or kinetic with intervals of at least 20 seconds readings Note For the following performance the Gain on the Optics Test should be below 6 in normal read mode Endpoint or Kinetic with an interval lt 20 seconds Absorbance Measurement Range 0 000 to 4 000 Abs Accuracy Linearity Repeatability 0 000 to 2 000 Abs 1 2 000 to
114. lect Reader System Test to verify that the light levels and electronic noise at all set wavelengths fall within factory acceptance criteria e Select Universal Test Plate to run the calibration plate test to confirm the alignment repeatability and accuracy linearity of the reader The report will also contain the part and version numbers of the software loaded on the unit e Perform a wavelength scan to confirm wavelength accuracy of the reader From the Main Menu press the soft key that corresponds to UTIL to access the Utility Options Menu The Select Utility Option screen will appear Selecting UTIL from the Main Menu e Select TEST to advance to the Select Test screen Choosing a performance test to run e Select SYSTEM to verify that the light levels and electronic noise at all wavelengths fall within factory acceptance criteria e Select CHKSUM to instruct the reader to compare the software to the internally recorded checksum values to ensure that the programming has not been corrupted In addition use this option to view the part numbers and versions for the software currently installed on the reader This information is needed when contacting Bio Tek for technical assistance e Select CALPLATE to run the calibration plate test to confirm the alignment repeatability and accuracy linearity of the reader PowerWaveX Operator s Manual 4 3 4 4 SYSTEM TEST The System Test confirms that the l
115. lerances during the assay Incubator Failure Error Data Bits Zone Error 7 3 Temperature out of range 6 2 Temperature out of range 5 1 Temperature out of range 4 0 Temperature out of range 3 3 Thermistor out of range or shorted 2 2 Thermistor out of range or shorted 1 1 Thermistor out of range or shorted 0 0 Thermistor out of range or shorted Examples Bit 6 Bit 0 Ar Error code 1540 40 hex 0100 0000 Zone 2 temp out of range unknown reason A D failure Error code 1544 44 hex 0100 0100 Zone 2 temp out of range due to zone 2 thermistor bad Error code 15FF gt FF hex 1111 1111 All zones temp out of range due to all thermistors bad Note Any failure indicates a need to run a System Test and review of the printout or KC4 screen Error Codes Displayed Error Potential Cause ERROR 1600 Computer control assay definition error Assay programming via computer control has tried to define an invalid assay sequence or parameter Displayed Error Potential Cause ERROR 1700 Kinetic interval too short for selected options ERROR 1800 Too many kinetic intervals programmed ERROR 1900 Memory allocation failure ERROR 1A00 Store curve error ERROR 1B00 Get curve error ERROR A200 Reader function already in use ERROR A300 Device not available ERROR A400 Failed code checksum on power up ERROR A500 Power good error power dropped below safe level ERROR A600 Quick fl
116. ll ASCII character strings representing numbers or names are transmitted most significant digit or letter first Data values not indicated as ASCII are treated as binary integers and are transmitted least significant byte first Lower wavelength limits shown are for the PowerWaveX 200 only The PowerWaveX 340 is limited to wavelengths above 340 nm and will not accept lower values The following section describes the supported computer control command set STORE PLATE CARRIER A This command causes the plate carrier to move inside the instrument host valid limits response A no arguments lt ACK gt status string 5 PRESENT PLATE CARRIER J This command causes the plate carrier to move back outside the instrument where it can be loaded with a microplate host valid limits response y no arguments lt ACK gt status string 5 PowerWaveX Operator s Manual B 7 CONFIGURE WAVELENGTHS M B 8 This command downloads the wavelength setup table for use in future read cycles host valid limits response M lt ACK NAK gt data bytes 1 3 Ist wavelength 200 999 ASCII 4 comma separator 5 24 remaining wavelengths commas lt ETX gt status string 5 Each wavelength is represented by xxx 3 ASCII digits followed by a comma Six wavelengths must be sent 24 bytes followed by ETX after the last comma If a wavelength is not to be specified i
117. llowing instructions briefly describe how to read a plate using KC4 Refer to KC4 s Help system and User s Guide early and often to learn how to create protocols assign well identifiers read plates print reports and more To read a plate using KC4 1 Select Data New Plate 2 If prompted to select a protocol select Empty Protocol and click OK If not prompted select Protocol New or use KC4 s Protocol Wizard to step through protocol creation 3 Select Protocol Reading The Reading parameters dialog will appear 4 Select a Reading Type of Endpoint Kinetic or Spectrum 5 Define the wavelength s at which the plate will be read 6 Select a Plate Geometry of 8x12 96 well plate or 16x24 384 well plate 7 Define other reading parameters as necessary Click the Help button for assistance 8 When complete click OK 9 Select Data Read Plate The Plate Reading dialog will appear 10 Enter any comments place the plate on the carrier then click Start Reading to begin the plate read e The plate will be read and then the raw data results will display in KC4 e To analyze manipulate or print results protocol parameters should be defined Refer to KC4 s Help system or User s Guide for instructions PowerWaveX Operator s Manual B 3 Controlling the Reader with KCjunior The PowerWaveX can be operated using a computer running Bio Tek s KCjunior software Follow the steps below 1 Power off the
118. logical assays Automatically stores results for the last 8 plates Spectral scanning using on board software with report of peak OD and wavelength at peak OD Introduction Package Contents e Microplate scanning spectrophotometer e Power cord e Operator s Manual and Warranty Registration Card PN 7261014 e Assay Reference Guide depending on the model PN 7271006 e Dust cover PN 7342066 e Parallel cable PN 71072 e Serial cable PN 75053 e PowerWaveX Select model only Allen wrench PN 49329 Optional Accessories e Service Manual for all PowerWaveX models PN 7261016 e Universal Test Plate PN 7260522 e Bio Cell M for 1 cm wavelength readings PN 7272051 e Bio Cell adaptor plate for containing up to eight Bio Cells PN 7270512 e Installation Operational Performance IQ OQ PQ package PN 7260219 PowerWaveX Operator s Manual Specifications e Microplates All models accommodate standard 96 well microplates The Select model also accommodates 384 well microplates 111 Note Testing is recommended with 384 well plate types to determine compatibility The Select was tested with Greiner and Costar 384 well plates and with Nunc black sided clear bottom 384 well plates and results meet specifications e Speed of Reading The actual plate read time is dependent on the method of reading gt In Normal read mode the reading speed is variable with the absorbance of the samples If a sam
119. lose three steps PROBABLE CAUSE e Belt slipping caused by incorrect tension loose motor pulley or loose belt clamp e Defective motor drive circuit or Main PCB Error Codes e In the wavelength select case failure to verify home correctly usually indicates an alignment problem e Anti backlash spring on the monochromator has become free of the mounting post e A defective flash lamp Displayed Error Potential Cause This error indicates that the optic system detected saturation conditions at the wavelength selected during the read PROBABLE CAUSE e Optic system is defective Displayed Error Potential Cause LAMBDA 1 Gain out of range ERROR 0602 LAMBDA 2 Gain out of range LAMBDA 3 Gain out of range LAMBDA 4 Gain out of range LAMBDA 5 Gain out of range LAMBDA 6 Gain out of range Errors 0601 0606 indicate that the gain for a specific wavelength is out of the range necessary to ensure the filter s performance to specifications PROBABLE CAUSE e A defective flash lamp misaligned optics or defective analog board e A defective order sorting filter PowerWaveX Operator s Manual C 3 C 4 Displayed Error Potential Cause ERROR 0700 Reader failed noise test The last digit indicates the channel 0 8 where 0 is the reference channel The reader noise test checks the DARK current signal level for stability Dark current is measured with the light blocked at maximum measurement channel ga
120. lt This test confirms the reader s ability to read samples accurately in different plate positions Each channel is compared to at least one other channel in this test A to H B to G C to F and D to E This ensures that all channels have comparable performance Performance Verification Qualification Tests Wavelength Accuracy The C6 filter should be scanned between 580 and 590 nm in 1 nm increments using KC4 or KCjunior software If neither KC4 nor KCjunior is available select six wavelengths at 1 nm increments near the expected peak The wavelength of the maximum absorbance should be compared with the wavelength written on the sheet supplied with the test plate see Figure 4 4 below The accuracy of the wavelength should be 3 nm 2 nm instrument 1 nm filter allowance MULTIBAND CALIBRATION FILTER IN LOCATION C WAVELENGTH nm TYPICAL ABSORBANCE SPECTRUM DIDIMIUM GLASS V 10 WAVELENGTH OF PEAK IN THE INTERVAL BETWEEN 580nm AND 590nm S87 Figure 4 4 Sample data sheet showing wavelength of peak in the interval between 580 and 590 nm For example If the data sheet value is 587 nm and the allowable range is 3 nm and if the reader reports a peak value of 590 nm then the reader meets its specifications If the reader reports 591 nm then the reader does not meet specifications PowerWaveX Operator s Manual 4 13 Liquid Testing Liquid testing tests the reader in ways th
121. lt motor gt FLASH MISS ERR 2500 motor went by flash location too soon XY LIMIT ERR 2600 physical limit exceeded for area scan request PANEL METHOD ERR 2700 lt assay gt method doesn t match first panel assay MOTOR TIMER ERR 2800 lt motor gt timer not available VREF ERR 2900 voltage reference failed lt test type gt PLATE JAM ERR 2A00 lt motor gt didn t find middle sensor Test Type Codes lowest digit in returned error code FAIL POWER 5V T SV power failed FAIL POWER 24V e 24V power failed Motor Codes lowest digit in returned error code Carrier X Axis 0 Filter Wheel 15 Robotic Door 2 R instruments only Monochromator 3 Carrier Y Axis SA Select only 2 on HT Incubator Codes second lowest digit in returned error code Range Error 0 Thermistor Error TP A D Error 2 Affected zones are encoded in the lowest digit returned one bit per zone Data Flash Codes 2 lowest digit in returned error code Readback Error 0 data read back didn t match data written Copy Error a final data readback didn t match original passed in A D Device Codes lowest digit in returned error code Absorbance measurement ly Fluorescence measurement 2 Incubation measurement EH Voltage reference Y PowerWaveX Operator s Manual B 23 B 24 Computer Control Appendix C Error Codes This chapter describes Error Cod
122. mal operation install the PowerWaveX on a level surface in an area where ambient temperatures between 18 C 65 F and 40 C 104 F can be maintained The reader is sensitive to extreme environmental conditions Conditions to avoid are e Excessive humidity Condensation directly on the sensitive electronic circuits can cause the instrument to fail internal self checks e Excessive ambient light Bright sunlight or strong incandescent light can reduce the linear performance range and affect the instrument s readings e Dust Optical density readings may be affected by extraneous particles such as dust in the microplate wells A clean work area is necessary to ensure accurate readings Unpacking and Repackaging the Instrument Important Keep the shipping cartons and the packaging material for the carrier s inspection If the reader is shipped to the factory for repair or replacement it must be carefully repackaged using the original packing materials Shipping with improper packaging materials may void your warranty If the original packaging materials have been damaged replacements are available from Bio Tek 1 Carefully open the top of the shipping container and remove the dust cover See Figure 2 1 The box will include a power cord Operator s Manual serial cord and optionally a Service Manual 2 Remove the top foam from the reader 3 Lift the reader out of the bottom foam and place it on a level surf
123. mid level solution into the wells of columns 3 and 4 of the microplate Finally pipette 150 ul of the low level solution into the wells of column 5 and 6 of the microplate Read the microplate using Normal mode single wavelength at 340 nm no blanking or blank on air Repeat the read four times for a total of five reads PowerWaveX Operator s Manual 4 23 Repeatability Test A 1 The plate data can be exported to a Microsoft Excel spreadsheet using KC4 or KCjunior M The mathematical computations described below may then be performed and the template kept for future data reduction 2 Calculate the mean and standard deviation for the five readings of each well 3 For each mean calculate the allowed deviation using the repeatability specification for a 96 well format of 1 0 0 005 Abs 4 The standard deviation for each set of readings should be less than the allowed deviation For example Absorbance readings of 0 802 0 802 0 799 0 798 and 0 801 will result in a mean of 0 8004 and a standard deviation of 0 0018 The mean 0 8004 multiplied by 1 0 8004 0 010 0 008 which when added to the 0 005 0 008 0 005 0 013 which is the allowable deviation Since the standard deviation is less than this value the reader meets the test criteria Linearity Test B 1 Obtain an average mean for each concentration by averaging the mean values for each well that were obtained above 2 Pe
124. n an assay is 100 The printed report shows up to 100 points and includes the Peak OD and wavelength at Peak OD PowerWaveX Operator s Manual 3 9 DELAY FIRST READ The Delay First Read option can be used to allow for sufficient time for microwell contents to settle or to incubate a plate before reading Delay in first read entry e Use the NUMERIC keys to enter the time in minutes and seconds The maximum delay time is 59 minutes 59 seconds 59 59 INCUBATION TEMPERATURE READER DEPENDENT Certain reader models are equipped with heaters to perform plate reads at elevated temperatures see Chapter 1 Introduction for information on these models The Incubation Temperature screen is used to enable incubation for this assay and to set the temperature Incubation Temperature entry e Select AMBIENT to read the plate at room temperature Note See Cooling Fan in Chapter 2 for information on lowering the ambient temperature by turning on the internal fan e Select TEMPERATURE to read the plate at an elevated temperature e If TEMPERATURE is selected use the NUMERIC keys to enter the incubation temperature The valid range is from 25 to 50 C Incubation performance is specified from 4 over ambient to 50 C 3 10 Operation WAVELENGTH S Use the Wavelength selection screen to specify whether the plate will be read at one SINGLE or two DUAL wavelengths
125. nd describes its hardware and software features and technical specifications Instructions on how to contact Bio Tek for Technical Assistance are included on page 1 8 Introducing the PowerWaveX Bio Tek s PowerWaveX is an eight channel reader assay system for research and development and in vitro diagnostic use The reader can serve as a stand alone system or as an integral part of a PC based system using Bio Tek s KC4 or KCjunior software packages The PowerWaveX is a spectrophotometer that has the capability of spectral scanning using a monochromator This allows the user to perform endpoint and kinetic analysis in 96 well microplates using any wavelength between 200 and 999 nm or 340 and 999 depending on the model thereby achieving the maximum absorbance for any sample The reader features superior optical specifications with an extended dynamic range of up to 4 000 absorbance units The instrument s on board processor 2 X 24 character LCD screen and membrane keypad allow easy definition and management of assay protocols templates formulas and data Results can be output in a printed report format or exported for use in a variety of microplate based data manipulation applications Variations Bio Tek s PowerWaveX may be configured with all or selected options e PowerWaveX Select models support the reading of 96 and 384 well plates e TY model instruments have a 4 Zone incubation chamber that controls
126. ono cnn r non cnn anna rre B 5 Controlling the Reader Using Serial Protocol oooononcccconociconnccccnnnoccnonocannnnnrrn cc nnnonnn rn B 6 Computer Control Command Set ococcccinocicinoccccnononcnnnonnncnn non n nc canon nnrnnnnn nn nan rr rn rr B 6 Using the Stop Key to Halt Plate Scans and Reads oooooconoccccconoccconoconcnnnoncncnanancnnnnnnnnos B 20 Status String FOMMAL cscs es ceke Nes decease i B 21 Appendix Cr Error Codes iicie tcicccicesccsetecsccccecsecesscssensscedesstecscctenccteseceteceses C 1 Appendix D Sample Reports occccccccccccccccccccccnccncncnonenonnnonnnnnnnnnenenennnos D 1 Preface List of Illustrations Figure 2 1 Figure 2 2 Figure 2 3 Figure 2 4A Figure 2 4B Figure 2 5 Figure 3 1 Figure 3 2 Figure 3 3 Figure 3 4 Figure 3 5 Figure 4 1 Figure 4 2 Figure 4 3 Figure 4 4 Figure 4 5 Figure 4 6 Figure D 1 Figure D 2 Figure D 3 Figure D 4 Figure D 5 Figure D 6 Figure D 7 Figure D 8 Unpacking and Repackaging the Reader ooonoonnonccniconocnconnconccnnconoconcnn nono nconnonnnonnnnos 2 2 Removal of the Carrier Retention Screw and Accompanying Washers from the PowerWaveX Solo e258 apona e aaa ill il 2 3 serialand Parallel Confectors triciclo an Sect orate Seman incio au 2 6 Voltage Selector Board and Fuse Holder ccceccceccesceseeeseceeeceeeeeeeseeeeeeeeeeereneens 2 12 Selecting Fuse Position KAN waste hee Boon Ree 2 12 Options Availa
127. opriate error flags in order for the user to determine their validity Because there have been no limits applied to the raw absorbance data all information exported via computer control must be analyzed completely Warning Unspecified Use Failure to operate this equipment according to the guidelines and safeguards specified in this manual could result in a hazardous condition PRECAUTIONS The following precautions are provided to help you avoid damaging the system Caution Service The system should be serviced by authorized service personnel Only qualified technical personnel should perform troubleshooting and service procedures on internal components Caution Environmental Conditions Do not expose the system to temperature extremes Ambient temperatures should remain between 18 40 C System performance may be adversely affected if temperatures fluctuate above or below this range PowerWaveX Operator s Manual v vi Caution Sodium Hypochlorite Do not expose any part of the instrument to Sodium Hypochlorite solution bleach for more than 30 minutes Prolonged contact may damage the instrument surfaces Be certain to rinse and thoroughly wipe all surfaces Caution Power Supply Only use the correct line voltage when operating the Automated Microplate Reader A four position line voltage select switch is used to adjust for different line voltages This switch is located on the power input module See the section Adju
128. ple measurement is less than 1 of the channel blank signal the reader will automatically go into an enhanced read mode where longer read intervals are applied for increased performance gt In Rapid read mode enhanced readings are not used therefore the reading speed is not variable Wavelength choice will also change read times slightly Each wavelength has a unique location within the monochromator and the different locations require varying amounts of time to position This effect is more noticeable in dual wavelength assays 96 Well Read Timing 630 nm 630 450 nm Normal Read Mode Single Dual Endpoint 23 to 31 sec 35 to 53 sec Rapid Read Mode Single Dual Endpoint 23 sec 35 sec Kinetics 14 seconds from Al to Al in rapid mode single wavelength 384 Well Read Timing 630 nm 630 450 nm Normal Read Mode Single Dual Endpoint 41 to 86 sec 71 to 141 sec Rapid Read Mode Single Dual Endpoint 37 sec 64 sec Kinetics 28 seconds from Al to Al in rapid mode single wavelength 1 4 Introduction e Optical Specifications A range 200 to 999 nm or 340 to 999 nm model dependent A accuracy 2nm A repeatability 0 2 nm A bandpass 5 nm e Optical Performance 96 Well Plate Reading The following specifications apply to 96 well flat or round bottom plates single wavelength endpoint or kinetic with intervals of at least 20 seconds readings Note For the following performance the Gain on the Optics test shou
129. ply for all decontamination or cleaning operations PROCEDURE Warning The bleach solution is caustic wear gloves and eye protection when handling the solution Do not soak the instrument keypad this will cause damage Wipe the keypad with a damp cloth 1 Turn off and unplug the instrument 2 Prepare an aqueous solution of 0 5 Sodium Hypochlorite NaCIO or bleach gt Be sure to check the NaClO of the bleach you are using this information is printed on the side of the bottle Commercial bleach is typically 10 NaClO if this is the case use a 20 1 mixture Household bleach is typically 5 NaClO if this is the case use a 10 1 mixture 3 Wipe down the carrier and all exposed surfaces of the unit with the bleach solution 4 Discard the used gloves and towels Cleaning Procedure Exterior surfaces may be cleaned not decontaminated with a cloth moistened not soaked with water or water and mild detergent Do not immerse instrument or spray with liquids A 2 Decontamination and Cleaning Appendix B Computer Control The PowerWaveX can be controlled either from the reader s front panel or from a computer connected to the reader via the computer s serial port This chapter describes the features of computer control and explains how to configure the computer to control the reader Overview With the computer control feature the user is provided even more power and flexibility For example the Power
130. port to a host computer The serial port also allows for computer control of the instrument and provides the means for downloading additional assay definition files to the instrument The PowerWaveX may be loaded with custom pre programmed assays and or with Open assays e The pre programmed assays allow you to easily select modify and run for example several common ELISA and important nucleic acid applications directly from the front panel e The Open assays contain standard default settings upon which assays can be programmed This chapter describes how to program Open assays on the PowerWaveX The custom pre programmed assays are described in Bio Tek s Assay Reference Guide PN 7271006 Important Screens and options throughout this chapter pertain to Open assays Pre programmed assays may not show all possible screens or options or may show custom screens not described here Please refer to the Assay Reference Guide for details on pre programmed assays Front Panel LCD Softkeys 4 Press a key to choose a menu function SELECTING MENU FUNCTIONS 10 23AM 370 C EF INE REPORT UTIL CLEAR ENTER Main Previous OE STOP READ Figure 3 1 PowerWaveX Front Panel Menu functions Press the left and right arrows to move the cursor in the display Press Options to scroll through options or the alphabet Press Shift Options to scroll
131. r of times with one single or two dual wavelengths Three different types of data are available see Kinetic Analysis on page 3 15 gt For arate determination the data calculated is the rate of OD change After determining the maximum rate of change per well using linear regression the units are milliOD min gt The R squared value is an indication of how well the points fit the linear curve gt The Onset time indicates how many seconds it took to reach a user s predetermined OD value e During WELL SCANNING the Optical Density values are measured at different points across the well diameter Choose from 1 to 31 points total 15 left of center 15 right of center If you choose 1 then the plate is read as an endpoint assay The data out is equal to the sum of all of the OD values recorded for the well To find the average OD value for the region scanned divide the scanned OD result by the total number of scans For more information see Well Scanning on page 3 16 Spectral Scanning Well Scanning and Spectral Scanning are two different read methods Certain readers contain the pre programmed assay SPECTRAL SCAN number 00 This assay allows for the definition of start stop and step increment wavelength values For example if the Start Stop Step values are 200 600 10 nm respectively each well will be read at 200 210 220 600 nm The maximum number of wavelengths that can be specified for reading withi
132. rWave 340 w Incubator and Robotic Access Door PowerWavex w Incubator and Robotic Access Door PowerWavex Select w Robot Access Door PowerWavex Select w Incubator and Robotic Access Door OPEN PLATE ACCESS DOOR This command opens the robotic plate access door host valid limits response no arguments lt ACK NAK gt status string 5 CLOSE PLATE ACCESS DOOR This command closes the robotic plate access door host y valid limits response no arguments lt ACK NAK gt status string 5 Using the Stop Key to Halt Plate Scans and Reads Pressing the STOP key on the reader while a computer control initiated plate read or spectral scan is in progress causes the following to occur 1 The scan or read process is halted and all axes are returned to their home positions the plate is moved back outside where it can be accessed 2 The reader transmits a lt DLE gt character to the computer when the above process is completed 3 No more data is transmitted by the read or scan in progress nor is a status string Computer Control Status String Format Following execution of each command the PowerWave sends a status string back to the computer This string consists of 5 successive ASCII characters RS S1 S2 3 and ETX gt RS A record separator that marks the beginning of the status string gt 1 Always ASCII zero 0 gt S2 A single digit used as a reader fault or error co
133. ranslated as 1 234 OD Overrange readings are indicated by the 5 character string instead of a signed numerical value Checksum calculation starts with the first byte AFTER the data start code up through and including the lt Z gt data terminator code at the end of each wavelength s plate data The checksum is transmitted as an integer data byte not ASCII PowerWaveX Operator s Manual B 9 SET ASSAY DEFINITION V B 10 This command downloads to the reader a 170 byte assay definition table to be used in subsequent plate reads Note To minimize confusion all don t cares should be set to 0 host valid limits response y lt ACK NAK gt data bytes 1 don t care 2 7 assay name ASCII characters 8 30 don t care 31 encoded byte bits 0 3 Ox0F 0 bit 4 0x10 1 shake 0 no shake bit 5 0x20 1 shake before every read 0 before first read only bit 6 0x40 1 continuous shake 0 timed shake bit 7 0x80 1 use total kinetic time 0 use kinetic read count 32 number of scans per well for well scanning must be an odd integer from 1 to 31 33 49 don t care 50 55 000000 ASCII 56 read type 0 endpoint 2 kinetic 3 scan Note 3 indicates well scanning To perform spectral scanning use the amp SCAN PLATE command 57 59 don t care 60 61 kinetic interval seconds O 9999 62 63 kinetic read count 2 9999 64 66 don t care 67 69 first wavelength
134. re 3 1 for instructions on using the keypad e Although the reader does not require it consider entering a unique ID for each plate especially if reading multiple plates for the same assay e Ifthe internal bar code scanner option is installed the reader will automatically scan the plate bar code label and use that for the Plate ID Note Use caution when creating multiple Plate IDs The reader does not provide a warning when the maximum of 8 Plate IDs stored in memory is about to be exceeded If a 9th Plate ID is added it will overwrite the first Plate ID stored in memory PowerWaveX Operator s Manual 3 49 Enter Sample ID If the Enter Sample ID prompt is presented enter a starting sample identification number Entering the starting sample identification number e The valid entry range is from 0001 to 9999 e The software will automatically increment each subsequent sample identification by 1 e The sample IDs will be assigned in accordance with the plate map defined in the assay Enter Well Location If the assay specifies manual plate mapping and if Prompt for Sample Count is set to Yes under UTIL Y READ sample well locations can be defined at run time Peete UE lie asa Entering the sample well locations at run time e The sample well locations originally defined in the assay will be presented If desired use the keypad to enter new well location for each sample replicate e Press ENTER
135. rform a regression analysis on the data to determine if there is adequate linearity For example In Excel under Tools select Data Analysis and then Regression Prior to opening the regression analysis tool enter the expected results of the dilutions in a row of cells to use in the analysis Use the Regression Input box to enter the expected values as the Input Y Range and the mean absorbance for each concentration as the Input X Range Expected results Since it is somewhat difficult to achieve high pipetting accuracy when conducting linear dilutions an R Square value of at least 0 99 is considered adequate 4 24 Performance Verification Qualification Tests Maintenance of Robotic Units Use the following procedure for periodic lubrication of PowerWaveX robotic units 1 Remove the shroud 2 Lubricate the components shown in Figures 4 5 and 4 6 every six months or after 10 000 cycles according to the following instructions Use Bio Tek lubricant PN 66039 a Procedure Location A Lubricate the underside of the bracket B Lightly lubricate the shaft indicated Apply a heavy coating of lubricant to both sides of the motor shaft Run the motor C shaft back and forth through the motor to ensure that the internal drive nut is heavily lubricated Use the robotic motor adjustment procedure provided below to work the lubricant into the internal drive nut D Ensure that the mylar
136. rm alignment repeatability and accuracy which will also confirm linearity If a Test Plate is not available Liquid Test 2 can be utilized for these tests Preparation of Dilutions 1 Set up a rack containing 10 tubes numbered consecutively 2 Prepare a concentrated stock test solution insert using either Procedure A on page 4 15 or Procedure B on page 4 16 3 Create a percentage dilution series beginning with 100 of the original concentrated stock solution in the first tube 90 of the original solution in the second tube 80 in the third tube all the way to 10 in the last tube Use Class A volumetric pipettes for better accuracy 4 Dilute using amounts of the remaining 0 05 solution of deionized water and Tween 20 as shown in Table 4 4 Table 4 4 Test Tube Dilutions Tube Number E Volume of Original E 4 12 10 Solution ml Volume of 0 05 E AG Ka pa Ra Tween Solution ml Absorbance expected 2 0 1 8 A 6 W aE a 4 k 2 if original solution is 2 0 at 200 ul Plate Preparation 5 Pipette 200 ul of the concentrated solution from tube 1 into each well of the first column A1 to H1 of a new flat bottom microplate Costar 3590 is recommended Next pipette 200 ul from each of the remaining tubes into the wells of the corresponding column of the microplate tube 2 into wells A2 to H2 etc Note The choice of dilutions and the absorbance of the original solution can be varied Use Table 4 4
137. s into Installation Performance and Operational Qualification categories Recommendations for Achieving Optimum Performance e Microplates should be perfectly clean and free of dust or bottom scratches Use new microplates from sealed packages Do not allow dust to settle on the surface of the solution use microplate covers when not reading the plate Filter solutions to remove particulates that could cause erroneous readings e Although the PowerWaveX supports most flat U bottom and V bottom microplates the reader achieves optimum performance with optically clear flat bottomed wells e Non uniformity in the optical density of the well bottoms can cause loss of accuracy especially with U and V bottom polyvinyl microplates Check for this by reading an empty microplate Dual wavelength readings can eliminate this problem or bring the variation in density readings to within acceptable limits for most measurements e Inaccuracy in pipetting has a large effect on measurements especially if smaller volumes of liquid are used For best results use at least 100 ul per well in a 96 well plate and 25 ul in a 384 well plate e Dispensing solution into 384 well plates often traps air bubbles in the wells Dual wavelength reads will cancel most of these errors however for best results they should be removed by degassing the plate in a vacuum chamber prior to reading e The inclination of the meniscus can cause loss of accuracy in som
138. say specifies manual mapping prompts for information will include the locations for the sample wells e If running a custom assay typical prompts might include refer to the Assay Reference Guide for more information gt The number of samples gt Standard concentrations Assay ID Fill pattern Blank method First well location Replicate count for each well type Wavelength mode Vv Vv Y VV VY Y Report preferences etc 3 48 Operation Enter Number of Samples If the Enter Number of Samples prompt is presented indicate the number of sample groups on the plate The number of sample replicates is typically pre defined in the assay but if this is a custom assay you may also be prompted to enter the replicate count Entering the number of sample groups on the plate e Use the NUMERIC keys to enter the number of sample groups e The valid entry range is from 01 to the maximum number of wells remaining on the plate after any blank control or standard wells are mapped e If you enter a value greater than the number of empty wells remaining on the plate the reader will beep and automatically change the value to the maximum permissible number of samples Enter Plate ID If the Plate ID prompt is presented enter a unique plate identifier to be stored in memory with the assay name and absorbance data BEBE BREE Perea OO IAN IEA Entering a Plate ID e Use up to 10 alphanumeric characters See Figu
139. scrites dans le Reglement sur le brouillage radioelectrique edicte par le ministere des Communications du Canada User Safety This device has been type tested by an independent laboratory and found to meet the requirements of the following e Canadian Standards Association CAN CSA C22 2 No 1010 1 1992 Safety Requirements for Electrical Equipment for Measurement Control and Laboratory Use Part 1 General Requirements PowerWaveX Operator s Manual vil C Based on the testing described below this instrument bears the CE mark EMC EC DIRECTIVE 89 336 EEC e EN 50081 1 CLASS A Emissions The system has been type tested by an independent testing laboratory and found to meet the requirements of EC Directive 89 336 EEC for Radiated Emissions and Line Conducted Emissions Verification was to the limits and methods of EN 55022 The device is classified as EN 55022 Class A e EN 50082 1 Immunity The system was also tested and found to meet requirements for Electrostatic Discharge Susceptibility Radiated Susceptibility and Electrical Fast Transient Burst Susceptibility Verification of compliance was conducted to the limits and methods of EN 50082 1 1992 IEC 1000 4 2 1995 IEC 1000 4 3 1995 IEC 1000 4 4 1995 EN 61000 4 6 1996 EN 61000 4 11 1994 EC Directive 73 23 EEC Low Voltage Safety The system has been type tested by an independent testing laboratory and was found to meet the requirements of EC Directive 73 23
140. sented is determined by the report settings Matrix Column Curve Fit specified under UTIL gt OUTPUT e Select MAP to print a matrix showing the locations of the Blanks Standards Controls and Samples for a particular assay e Select ASSAY to print a plate map and a listing of all of an assay s settings such as wavelengths numbers of well types formulas and curve fit parameters e Select LIST to print a list of all assays name and number currently programmed in the reader Operation Results Report The reader stores the absorbance data for the 8 most recent plate reads Results reports can be generated for these plates if for example the data that automatically printed after the read needs to be printed in a different format or if the standard curve contains outliers that require editing apes eve yale rata eh a Ne eb ee eee er ee hele epee Choosing a plate for the Results Report e The most recently read plate is presented first showing the assay name the plate ID if one was entered and the date the plate was read e Press OPTIONS to see the next plate in memory e Press ENTER to select a plate and continue e Ifa standard curve was generated and EDIT STANDARD OUTLIERS was set to MANUAL in the assay definition the EDIT STD OUTLIERS prompt is presented otherwise the Print Results prompt is presented Editing Standard Outliers If a standard curve was generated and if EDI
141. so housed within the power input module The following instructions can be used to change the input voltage range Figures 2 4A and B illustrate the power input module Figure 2 4A Voltage selector board and fuse holder Figure 2 4B Selecting fuse position 1 Unplug the reader and remove the power cord 2 Use a small flat blade screwdriver to pop the fuse holder out of the power input module 2 12 Installation PowerWaveX Operator s Manual A small voltage select switchboard is located on the right of the power input module The switchboard which can be removed with needle nose pliers can be oriented four ways to select four different voltage input ranges Once the switchboard is removed text can be read on one side of the board This text identifies the switch positions The text is always facing the ON OFF switch when installed in the power input module The text matching the desired voltage range should be on the edge of the switchboard which goes into the power input module as the arrow on the board indicates The white plastic indicator will need to rotate around so that it fits into the correct groove on the switchboard Verify proper fusing before reinserting the switchboard and fuse holder Once the switchboard is reinstalled the white plastic indicator peg should line up with the fuse holder voltage indicator holes If the peg does not fit in the hole that indicates the voltage intended the switchboard
142. st 3 Procedure 1 5 Check the absorbance of a sample of either high level test solution created in Procedure A or B at 340 nm on the microplate reader This solution will have an optical density absorbance of approximately 0 700 to 1 000 This value is not critical but it should be within this absorbance range Adjust up by adding B NADH powder if low until the high level test solution is at least at the lower end of this range Do not adjust if slightly high Carefully prepare a mid level test solution by diluting 15 ml of the high level test solution with 5 ml of the Sigma PBS solution If using the 10X concentrate PBS solution you should mix one part of the concentrate with nine parts of deionized water to obtain a low level buffer similar to the Sigma PBS Then use 5 ml of this solution as the diluent This will be the mid level solution Carefully prepare a low level test solution by diluting 10 ml of the high level test solution with 10 ml of the Sigma PBS solution If using the 10X concentrate PBS solution you should mix one part of the concentrate with nine parts of deionized water to obtain a low level buffer similar to the Sigma PBS Then use 10 ml of this solution as the diluent This will be the low level solution Pipette 150 ul of the concentrated solution into each well of the first two columns Al to H1 and A2 to H2 of a flat bottom microplate Costar 3590 is recommended Next pipette 150 ul from the
143. sting Line Voltage Input Range in Chapter 2 for more details Caution Carrier Retention Screw The carrier retention screw must be removed prior to operating the device Preface Electromagnetic Interference and Susceptibility USA FCC CLASS A Warning Changes or modifications to this unit not expressly approved by the manufacturer could void the user s authority to operate the equipment This equipment has been tested and found to comply with the limits for a Class A digital device pursuant to Part 15 of the FCC Rules These limits are designed to provide reasonable protection against harmful interference when the equipment is operated in a commercial environment Like all similar equipment this equipment generates uses and can radiate radio frequency energy and if not installed and used in accordance with the instruction manual may cause harmful interference to radio communications Operation of this equipment in a residential area is likely to cause interference in which case the user will be required to correct the interference at his own expense Canadian Department of Communications Class A This digital apparatus does not exceed Class A limits for radio emissions from digital apparatus set out in the Radio Interference Regulations of the Canadian Department of Communications Le present appareil numerique n met pas du bruits radioelectriques depassant les limites applicables aux appareils numerique de la Class A pre
144. t compare the mean values for the other wells to their corresponding mean values with the well in the turnaround position Compare B1 to G12 C1 to F12 D1 to E12 El to D12 Fl to C12 Gl to B12 H1 to A12 A2 to H11 and B2 to G11 etc The difference in the values for any two corresponding wells should be within the accuracy specification for the instrument For example If the mean of well A1 in the normal position is 1 902 where the specified accuracy is 1 0 010 Abs then the expected range for the mean of the same well in the H12 position is 1 873 to 1 931 Abs 1 902 1 0 019 0 010 0 029 which is added and subtracted from 1 902 for the range If any set of well values is out of the expected range review the other three sets for the same channel pair Thus if Al and H12 are not within range of each other review the compliance of H1 to A12 A2 to H11 and H2 to A11 This will confirm that there is a problem in one of the eight read channels or indicate that the result of one set of wells was in error If any two sets of well values for a channel pair are out of the allowed accuracy range there may be contamination on or a problem with one of the lenses 3 Ifthe four corner wells are within the repeatability range the reader is also in alignment PowerWaveX Operator s Manual 4 2 LIQUID TEST 3 To verify operation of the PowerWaveX at 340 nm perform the Liquid Test 3 Procedure descr
145. t display showing the assay configuration software part number and version number Performance Verification Qualification Tests Calibration Verification It is considered good laboratory practice to periodically verify the calibration of the PowerWaveX Verification should be performed monthly using the tests in this section Universal Plate Test see below Liquid Tests see page 4 14 UNIVERSAL PLATE TEST The Universal Plate Test also referred to at the Calibration Plate Test confirms the alignment repeatability and accuracy linearity of the PowerWaveX An alternative method used to determine accuracy repeatability and linearity is the Liquid Test 2 described later in this chapter The Universal Test Plate PN 7260522 allows comparison of the reader s optical density measurements and mechanical alignment to NIST traceable values Accuracy linearity repeatability and alignment are tested Specific standard calibration values must be entered for each wavelength to be tested see Entering the Universal Test Plata Data on the following page The Universal Plate Test confirms the following Accuracy of the Optical Density readings the comparison of the optical density readings with those given with the Universal Test Plate insert will confirm the accuracy of the optical density readings at specific wavelengths Linearity of the Optical Density readings are confirmed by default if the optical density read
146. t must be replaced by 000 The wavelength setup table is stored in non volatile memory and only needs updating when a change in current wavelength configuration is desired After wavelength configuration a self test command sequence must be run to calibrate gains and generate self check information for the new wavelength configuration It is not absolutely necessary to configure and calibrate wavelengths for use but performance may be compromised slightly if this is not done The wavelengths stored are automatically calibrated during the instrument power up self test Computer Control READ PLATE S This command causes a microplate to be read according to the currently loaded assay definition table host valid limits response Ss no arguments lt ACK NAK gt Reader response protocol ASCII format status string 5 for each wavelength specified up to six wavelength start code 1 lt CR gt for each row for each column well within specified strip range 1 comma separator 1 sign 1 Hor data 4 1234 row terminator 2 lt CR gt lt LF gt data terminator 1 lt Z gt checksum 1 0 255 not ASCII status string 5 only if read error Absorbance data is returned in the form of a 5 character ASCII string with a sign character followed by four digits A decimal point is not included but should be inserted by the user after the first digit i e 1234 should be t
147. table field e g under H e Use the ALPHA and NUMERIC keys to change the Assay name e Use the OPTIONS key to sequentially advance the character positioned above the cursor The characters will cycle through the alphabet A Z with a space following Li e Use the LEFT lt 4 and RIGHT arrow keys to move the cursor to the previous or next editable field The cursor will wrap around the edit field e Use SOFT KEYS 1 2 3 and 4 to select a dash forward slash colon or space for inclusion in the assay name PowerWaveX Operator s Manual 3 7 Define METHOD To define read method parameters start at the Main Menu and select DEFINE select the assay press ENTER then select METHOD Define options screen The definable read method parameters include Endpoint Kinetic or Well Scanning Read Modes Delay first read Incubation parameters Wavelengths applied Shake parameters Kinetic analysis Number of points for well scanning Spectral scan Assay 00 only Note Some screens described in this section may not appear on certain reader models READ TYPE Use the READ TYPE screen to specify the reading method Endpoint Kinetic or Scan Read Type screen 3 8 During an ENDPOINT read the plate is read once with one single or two dual wavelengths One optical density OD value is reported for each well Operation e During a KINETIC read the plate is read a defined numbe
148. ter 2 Displayed Error Potential Cause ERROR 0D00 Calibration checksum error This error indicates that the stored checksum value for the calibration data does not match the actual checksum This indicates that the data in the quick flash memory has been corrupted probably due to an electronic defect Displayed Error Potential Cause ERROR 0E00 Wavelength not detected in reader s Lambda table Error 0E00 indicates that the specified assay wavelength is not in the Lambda table and must be programmed in Displayed Error Potential Cause ERROR 0F00 Wavelength signal is out of specified range This error indicates that the measurement frequency selected has a signal out of range The causes could be many Essentially the signal produced at a specific frequency is either too high or too low to allow performance within the specification The lamp could have gone bad or a part of the optics system could be out of alignment This can also be a flag for filters starting to degrade Error Codes Displayed Error Potential Cause ERROR 1000 Configuration data is missing This error indicates that necessary configuration data is missing from memory which probably means it was never downloaded or it was downloaded incorrectly Displayed Error Potential Cause ERROR 1100 Failed configuration checksum test This error indicates that the stored checksum value from the configuration data does not match the actual
149. the volume in the microwell The diluted solution should have ODs of half of the concentrated solution 2 Pipette 200 ul of the concentrated solution into the first column of wells of a new 96 well flat bottom microplate Costar 3590 is recommended A new microplate is required for this test as any scratches may cause variations in the turnaround reading 3 Pipette 200 ul of the diluted solution into the second column of wells 4 Read the microplate five times at 405 nm using normal mode single wavelength no blanking 5 Next turn the microplate around so that well Al is now in the H12 position and read the plate five more times 6 The plate data can be exported to an Excel spreadsheet using KC4 or KCjunior The mathematical computations described below may then be performed and the template kept for future data reduction Channel to Channel Variation 7 Calculate the means of the wells in columns and 2 in the normal plate position and in the turnaround position This test results in four comparisons of each channel to its corresponding channel two in column 1 and two in column 2 Compare the mean reading for well Al to its mean reading when in the H12 position Next compare the mean values for the other wells to their corresponding mean values with the well in the turnaround position Compare B1 to G12 Cl to F12 D1 to E12 El to D12 F1 to C12 G1 to B12 H1 to A12 A2 to H11 B2 to G11 etc The
150. tiON oooooonncnnnncccnonococonococcnnnonncnanonnnn nan cnn nnannnnnnno 3 21 Blank Map s 3 21 Blanking Constant coin iiaii iiid 3 22 Number of Blank EEA E 3 22 Blank LOCA iia 3 23 N umber of Standard su cccootecia geccha i iutchent vecnensiac cubes seedbaseeetesebteteenatee 3 23 Number of Standard Replicates 20 0 0 eee eeeeeeeeeeeeeeeeeneeeseeaeeeeeneeeeeneeeeeeaaes 3 24 Average NAS vi iii cdi 3 24 Standard CONCENTTAtiONS ooooocononcccnonocanononcncnanonnnc nono cnn nnnn cn naar nn nr naar nnn nan nnnnnnnns 3 25 Number Of Controls seserian ei shes 3 25 Control Type ui le i ee ee eee 3 26 Number of Control Replicates ooooooooccccnnnnninococcccnccccononancconcncnnnnnano nn nnncnnnannnns 3 26 Location Of ControlS uscar ayes cs atesleedasa ite eigdieete sh 3 27 Number of Samples aee dec caide ata ade tc ibe eeiteca adea aa 3 27 Number of Sample Replicates oooooooconncccnnnociconococinononcnonanancno nono nc nano nn nnnnnn no 3 27 Sample LOCAtON roo AE 3 28 PowerWaveX Operator s Manual xvii Calculation SUCIA id 3 29 Eomula Tp A See ie at tae Ma eb ceed 3 30 Formula EntiYiiiiininaritaa riadas 3 31 MATH sota td ten sde le ds ea 3 32 OTHER cos e tor o hee ae de o soto 3 32 MAP e e O E 3 32 FUNCTION cocos ii Dia 3 32 Validation Formulas coin cece cdes sence tee iie E este Eiai iaaiiai 3 33 Control and Blank Validation Formulas ooocoonnccccnnociconoccccnanonnnonanancnnnnncnnnnns 3 33 Examples socsiie Mer
151. tional verify A D signal saturated lt test type gt lt filter gt gain out of range reader channel failed noise test reader channel failed offset test read time channel lt filter gt dark out of range read time channel lt filter gt air blank out invalid lt assay number gt printer timed out failed calibration checksum test wavelength not found in table lt read filter gt channel lt filter gt signal out of range necessary configuration data missing failed configuration checksum test necessary calibration data missing lt motor gt not homed successfully incubator failure error code lt zone s gt computer control assay definition error interval too short for selected options too many kinetic intervals malloc failed store curve failure 1B00 1000 get curve failure A D calib STBY transition not detected Computer Control Non Fatal Errors continued RESULTS DATA ERR 1D00 results data error CLOCK ERR 1E00 error in clock communications OVERLAP ERR 1F00 bandpass overlap in filterset BARCODE ERR 2000 no valid barcode detected INVALID PARAM ERR 2100 invalid parameter value selected PMT ERR 2200 PMT test signal too high lt test type gt LAMP ERR 2300 lamp control failure lt test type gt SENSOR POS ERR 2400 test sensor position incorrect
152. to advance to the next replicate Operation BEGINNING THE PLATE READ When all required prompts have been responded to the Place Plate in Carrier prompt appears Final prompt before the plate read begins e Before reading the plate make sure that the printer is connected turned on and full of paper e Place the plate on the carrier then press the READ key to initiate the plate read e After the read is complete data reduction will perform Calculating Results and then the reports will be generated Generating Reports e To halt in read in progress press the STOP key Note If using the incubation option the reader will wait for the incubator to reach temperature before reading the plate PowerWaveX Operator s Manual 3 51 Printing Reports Reports are automatically generated after a plate has been read see Specifying Data Output and Reporting Options in Chapter 2 for information on selecting reports Results reports also can be regenerated manually by using the REPORT option from the Main Menu In addition Map Assay and Assay List reports can be printed Note See Appendix D for sample reports PERO ercer E Ae le BEBE RBBB EG ee Generating reports via the REPORT option from the Main Menu e Select RESULT to obtain an exact copy of results from the plate reading the 8 most recent sets of plate data are stored in memory gt The form in which the results are pre
153. ubstantial noise C 2 Displayed Error Potential Cause ERROR 0300 Carrier failed to find light beam ERROR 0301 Order sorting filter wheel did not find home ERROR 0303 Monochromator did not find home Errors 0300 0301 and 0303 indicate that a particular axis has moved to a point where the light beam from the optics is no longer detectable by the measurement electronics PROBABLE CAUSE e Carrier A loose belt loose motor pulley or defective motor drive may cause the carrier to ignore movement instructions e Order Sorting Filter Wheel Motor The filter wheel drive gear is loose or motor drive failure is impeding filter wheel movement e Flash Lamp The flash lamp is not flashing due to a defective lamp poor alignment or blocked optics Displayed Error Potential Cause Track carrier failed position verify Order sorting filter wheel failed position verify Robot lid lift failed position verify ERROR 0403 Wavelength select monochromator failed position verify ERROR 0404 Y axis failed position verify Power WaveX Select models only Errors 0400 0401 0402 0403 and 0404 indicate that an axis failed its position verify test The position verify test verifies that the axis returns to the opto or the zero order mono case in the same number of steps as it is expected to be away from home If the axis does not return home in the required number of steps the test fails Each axis is allowed to gain or
154. ud rate used for transmission is held in nonvolatile memory and can be changed by the user Other serial port parameters No Parity 8 Data Bits and 2 Stop Bits are fixed and cannot be changed The RS 232C serial port on the Power WaveX is configured as a DTE see the section Setting Up the Serial Port for Communications in Chapter 2 that is the unit is wired to look like a modem Data is received on Pin 3 the RX Pin and transmitted on Pin 2 the TX pin Computer Control Command Set A command from the computer to the reader consists of a single ASCII character and in some cases subsequent argument data Upon receipt of a valid command character the reader returns an lt ACK gt character Some commands also return response data to the host computer Upon completion of command processing the PowerWaveX transmits a status string to the computer While awaiting a command the PowerWaveX responds to nulls or other unexpected characters by clearing its input buffer and transmitting a lt NAK gt Therefore if valid commands are preceded by invalid characters they may be missed Refer to Table B 2 below for the ASCII Control Characters used in the computer control protocol Table B 2 ASCII Control Characters Used in Computer Control Protocol ASCII Hex Decimal Control Reader Code Code Code Code lt gt NAK Negative 15 21 AU gt acknowledge Record separator Co cameram wo fs fm B 6 Computer Control A
155. ula Entry screen appears Each formula can contain a TS Spaces are not necessary Formula entry screen the Formula l prompt disappears to provide more spaces e After a moment the FORMULA 1 prompt disappears and the formula can be entered Use the opt formula ions found under MATH OTHER MAP and FUNCTN to build the gt To cycle through the available Math Other Map or Function options co ntinue to press the appropriate SoftKey For example press the MATH Softkey several times to see etc When the desired option appears press the RIGHT ARROW key to select it and advance to the next editable field gt Press the LEFT ARROW key to move the cursor to the left gt Press CLEAR to delete the item above the cursor gt When a formula is complete press ENTER to continue e Select MAT H to insert a mathematical symbol such as or lt e Select OTHER to insert an opening or closing parenthesis or logical operators A e Select MAP ND or OR to insert a well ID such as BLK x or NC 1 e Select FUNCTN to insert a mathematical function such as LOG or SORT Note The reader software checks the formulas for errors during data reduction A syntax error in a formula will result in a Token Error on results reports PowerWaveX Operator s Manual 3 31 MATH The following mathematical symbols can be used in formulas Ad
156. urn to the Main Menu See Adjusting the Reader s Wavelength Table Settings in Chapter 2 for instructions KINETIC READ PARAMETERS If the Kinetic read type was specified a few accompanying parameters must be defined including kinetic method reading interval and number of reads Kinetic reading method selection screen e Select TOTAL READS to read the plate a specified number of times e Select DURATION to read the plate for a specified length of time Kinetic Interval Regardless of the kinetic reading method selected Total Reads or Duration a Kinetic Interval is required This is the length of time in hours minutes and seconds between each kinetic read Kinetic Interval entry screen e Use the NUMERIC keys to enter the interval e The valid entry ranges are 0 1 hours 0 59 minutes and 0 59 seconds e Note Ifthe kinetic reading method is DURATION the Interval Duration combination must result in 2 to 40 total kinetic reads 3 12 Operation Total Number of Kinetic Reads If the TOTAL READS kinetic reading method was selected specify the total number of kinetic reads Eeee ele aerea e Le Kinetic Reads entry screen e Use the NUMERIC keys to enter the total number of reads to perform e The valid range is 2 to 40 reads Kinetic Duration If the DURATION kinetic reading method was selected specify the total length of time during which readings will be taken Kinetic Dur
157. urned alone with no wavelengths Calibration is not absolutely necessary with each plate but is recommended in scans where absolute accuracy is a requirement Due to the fact that even row detectors are offset left by one column the reader uses two passes to collect data when an entire strip is selected to be scanned During the first pass only the wavelength being scanned is returned Absorbance data is returned for each wavelength for the entire strip during the second pass The first pass terminator is always returned even if no first pass is necessary If more than one well or strip is to be scanned the series option can be used to make the process more efficient If a sequence of strips is to be scanned using the series option they must be contiguous and selected in sequence from left to right since first pass data for the next strip is collected during the second pass for the currently selected strip The last strip selected last scan then requires no first pass Calibration is only performed if the single or first scan option is selected and remains valid through the remaining wells or strips in the series Absorbance data is returned in the form of a 5 character ASCII string with a sign character followed by four digits A decimal point is not included but should be inserted by the user after the first digit i e 1234 should be translated as 1 234 OD Over range readings are indicated by the 5 character str
158. ved back outside where it can be accessed 2 The reader transmits a lt DLE gt character to the computer when the above process is completed 3 No more data is transmitted by the read or scan in progress nor is a status string SET PLATE GEOMETRY This command selects a microplate geometry to be used with the currently loaded assay definition host valid limits response f lt ACK NAK gt data bytes 1 geometry 2 or 3 2 96 wells 8 x 12 3 384 wells 16 x 24 If no Set Plate Geometry command is sent after the assay definition table the default 96 well plate is assumed it is recommended however that you issue this command even if using the 96 well plate If only a range of strips is desired the Set Strip Range command must be sent AFTER the Set Plate Geometry command B 12 Computer Control SET STRIP RANGE This command selects a range of adjacent strips numbered plate columns to be read host valid limits response lt ACK NAK gt data bytes 1 first strip 1 24 1 last strip 1 24 status string 5 By default the instrument is initialized to perform full plate reads and automatically reverts to this default when a new assay definition table is loaded either from the front panel or by computer control The strip range is also reset when the Set Geometry command is sent GET MINIMUM KINETIC INTERVAL This command provides a means for the user
159. werWaveX Select PowerWaveX Operator s Manual 2 3 AFTER UNPACKING VERIFY PERFORMANCE Before using the PowerWaveX for the first time verify that it is operating properly by turning it on Each time the reader is turned on it performs a system self test If the self test completes successfully the reader is ready for use If the test fails note any error codes and contact Bio Tek Refer to Chapter 4 Performance Verification Qualification Tests for a recommended test schedule which includes Installation Performance and Operational Qualification Tests BEFORE REPACKAGING THE INSTRUMENT 1 Decontaminate the reader prior to shipping see Appendix A Decontamination and Cleaning 2 For PowerWaveX Select models replace the carrier retention screw and its accompanying washers See Figure 2 2 for an illustration 3 Once the reader is clean pack it in its original shipping box using original packing materials This shipping system was designed to be used no more than five times If the container is damaged and or has been used more than five times contact Bio Tek for a new set of shipping materials PN 7263004 Adjusting the Reader s Wavelength Table Settings The PowerWaveX has an internal wavelength table that stores six user definable wavelength values When defining an assay these six wavelengths are presented as selectable options These values can be easily viewed or changed e From the Main Menu select UTI
160. with the inclusion of a pre defined Transformation Variable see TVAR below Simple Transformation Formulas Simple transformation formulas are typically applied to all wells on the plate For example e To divide the OD in each well on the plate by 2 and then multiply by 100 the formula is OD 2 100 e To raise the OD in each well to the power of 10 the formula is ODPWR10 Transformation Variable TVAR For more complex transformations a Transformation Variable TVAR can be defined for use within a transformation formula This variable defines the scope of the transformation whether to apply the transformation to all of the wells on the plate OD or to just the sample wells SMP Choosing the scope of the transformation PowerWaveX Operator s Manual 3 35 e If SMP is chosen gt The transformation formula will be applied to the sample wells only gt SMP and any other well identifiers BLK PC NC STD etc defined will become available as MAP options when building the transformation formula gt Example The assay plate map contains 2 NC wells and 2 PC wells The remainder of the map is filled with samples The assay data reduction requires that the mean of the NC be subtracted from all the samples on the plate The transformation formula is SMP NC x e If OD is chosen the formula definition screen will appear so that you can define a formula for use within the transformation formula gt

PowerWaveX Operator`s Manual - Bio-Tek

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