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μQuant™ - Bio-Tek

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1. iii LISTIOlFIGUPCS ni otra TI LEUTE viii SCOTT Ss Pc E ix Contacting BIOTek Instr msents Minis X Customer Service and Sales 5o Em EPI EbUs albla X SSE NICS eMe d oat lada X European Coordination Center nn ds X Document CONVENTION S A xi REVISION ESTOY Zosssutmat dus senate Sapa d lOO aia Xil Intended Use Statement ssseeeeerliieeeesee a rr XV Jual COMPE stad quxtacberad sa APO van bU P LCDP NIME euntayenesteohiacoemeanasataneniaies XV Specimen Preparatlobiissseseueaxto due vara bk AAA XV Repackaging and Shipping ccsssseeeeeee eene nnn nnn xvi Wang fer e D T XVI Hazards and PrecaltOMSs std clipes xvii HAZ al GS aynata o sb da xvii PrFECALUON Sd Dm xviii Directive 89 336 EEC Electromagnetic Compatibility ccceeeeeeeeeeeeeees xix Directive 73 237 EEG LOW Voltage ini A xix Directive 2002 96 EC Waste Electrical and Electronic Equipment xix Electromagnetic Interference and Susceptibility esee XX User Sata veo dias ERR eU sateen ena VneRV M Edu aac NM eeu n II dE XX Salebv SS VITIDOIS sissies outers nes ddp sees ene ean ao ae ae ae ee eee xxi EntrOOdUCctiOD sussetsxausadEPEEEPRE KE EEUREEFEPE RENE MEME ENSE NM NN EE RUN NE NM Ud D Introducing the uQuant O 2 Quality CONTO leresna d a e ar c hio ira a a Mc nte nia 2 Hardware Features stories 3 SoftWare Features scsi oi dc ic e LI DUAE 3 Package
2. oa O RS O O IS IS O O IS E ISID SID y D sip Sipe E tt qp 8 WW Pp p o E jsms sms __ py y F PE PC LL Po Pp p LG NC INC P H SMP s j Jj jy Jj o Map Direction ACROSS Rep Direction DOWN sb d 8 4 18 7 7 js J9 0 n 12 Seo sus sms jos re A ET a femi stb2 sros Fermi sros pre c tam se 1 1 1 BioTek Instruments Inc Define Method Map Formula and Curve 51 Start Mapping at Well Location The START MAPPING AT WELL LOCATION screen is only shown if automatic mapping is selected This option allows you to enter the location of the well that will be the starting point for automatic mapping R T MAPPING WELL LOCATION e Use the numeric and alpha keys to enter a letter or number at the cursor location For any well location only the alpha keys are active for the first character and numeric for the second and third characters The valid entry range is from A01 to the last well on the plate depending on the plate type and the number of blanks standards controls and or samples defined in the assay e Press ENTER to save the well location and continue uQuant Operator s Manual 52 Chapter 4 Operation Blank Map This option allows you to select which blanking method to apply to the assay The blanking options AIR FULL and CONSTANT ROW and COLUMN and
3. Baud Rate Data Bits Parity Stop Bits EOT Character 26 0014 Figure 32 Reader Setup dialog 4 Define the communication parameters 5 Click Test Communications to establish communication with the reader then click OK If the test fails follow the instructions provided by KCjunior UQuant Operator s Manual 186 Appendix B Using 384 Well Plate Geometry 6 Click the Read Plate button in KCjunior s main screen The Read Plate Dialog will appear If desired enter the Results ID and Plate Description Read Plate Dialog E X neste Read Plate uQuant Simulation D ata Protocol Mame Read From File SAM PLE Aflatoxin Results ID esults Help Plate Description Figure 33 Read Plate dialog 7 Click Read Plate to open the Protocol Definition dialog Define the Read Method parameters gt Selecta Read Method Type Define the Wavelength s at which the plate will be read Set the Plate Geometry to 16 x 24 Vv vV Y Define other reading parameters as necessary Click the Help button for assistance When you have finished defining the parameters click Validate Read Method if you would like KCjunior to verify the defined parameters If all parameters are valid you will receive confirmation The read method is valid for the configured reader If any are invalid KCjunior will provide instructions for selecting the correct parameters BioTek
4. gt To cycle through the available MATH OTHER MAP or FUNCTION Options continue to press the appropriate SOFT KEY For example press the MATH SOFT KEY 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 Press CLEAR to delete the item above the cursor gt When a formula is complete press ENTER to continue Select MATH to insert a mathematical symbol such as or lt Select OTHER to insert an opening or closing parenthesis or logical operators AND or OR Select MAP to insert a well ID such as BLK x or NC 1 Select FUNCTN to insert a mathematical function such as LOG or SQRT 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 BioTek Instruments Inc Define Method Map Formula and Curve 65 MATH The following mathematical symbols can be used in formulas Addition EN Subtraction ETT Multiplication OTHER The following additional symbols can be used in formulas 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 Greater than or equal to lt Less than or equal to FU
5. Note Do not discard the plate you will use it for the Alignment Test Print the raw data or export it to an Excel spreadsheet using Gen5 KC4 or KCjunior software If you are exporting the data to an Excel spreadsheet perform the calculations described below and on the following page and keep the spreadsheet for future tests Calculate the mean absorbance for each well and average the means for each concentration Perform a regression analysis on the data to evaluate linearity For example using Microsoft Excel In a spreadsheet create two columns labeled X and Y Enter the actual absorbance values in column X Enter the expected absorbance values in column Y BioTek Instruments Inc Liquid Testing 125 Select Tools gt Data Analysis Regression Identify column X as the Input X Range and column Y as the Input Y Range and then click OK to perform the analysis Note If the Data Analysis command is not available on the Tools menu you may need to install the Analysis ToolPak in Microsoft Excel Consult Excel s help system for assistance Expected Results Since it is somewhat difficult to achieve high pipetting accuracy when conducting linear dilutions an R Square value greater than or equal to 0 990 is considered adequate Repeatability Test 1 Calculate the mean and standard deviation for the five readings taken above at each concentration Only one data set needs to be ana
6. This error indicates one of the following situations e During a spectral scan the flash on value minus flash off lt 500 normal mode or lt 500 calibrated resets 8 for sweep mode During spectral scan the blanking on air uses minimal flashes to test the light performance The following ratio is used to determine the performance of the lamp Reference channel blank for wavelength flash on reference data corrected reference dark offset The ratio will fail if lt 0 5 or gt 2 0 Probable Causes Lamp failure alignment or lamp power supply Reference channel analog PCB or absorbance channel analog PCB or the cable in between The order sorting filter wheel is jammed and not aligning the correct filter to line up with the light path the filter is degraded and not passing enough light energy or the filter is blocking the light Damaged optic spray BioTek Instruments Inc Error Codes 159 General Errors Continued OFO1 Reference channel correction outside limits See Delta Air Dark OFO6 on self test See criteria below This error indicates one of the following situations e During system test the Delta was lt 500 for one of the wavelengths e This error indicates that during sweep mode only the blanking on air uses one flash to test the light performance The following ratio is used to determine the performance of the lamp Reference channel blank for wavelength flash on referen
7. sess mmn nn 28 Before Repackaging the Instrument eese nmm 28 Repackaging the uQuant and Its Accessories eeeeeeennn nnn 28 Mounting th SHIPPING IBIOGK sados 29 Packing the Reader and Accessories csse nennen nnn 32 Preparing the Shipping Container eese nennen nnn n 33 BioTek Instruments Inc Contents V Operacion aia D OUGHT Front Panels air 36 Sg Te TD 38 SO SS il OCA UD a biu e rb D PIE IE M E EE LM cM EIE CE 38 Mamen CP erc CES 39 DSTI M 40 Selecting an Assay to Define sias anesad asa ak ress alc C ais 41 Editino the Assay NaMe wiiatsaudccsdaavwplesesagarwe ears novelas iA i aaia 42 Define Method Map Formula and Curve cccccccccccnncnccnannnnncn nr 43 DEMNI dap TT 43 DENNO MAP eT TET 47 Denninhg FORMULA cosida 62 BA Sete a wetet ahaa sea Et oU Eb dE PERS 74 Reading a MIChO Pl Ale ich a anio 83 Selecting an ASSay to RUN ias 84 PENA REDORTO cee agen ioe ios 88 Editino Standard OUTS vosmtuv brani pedes mia a aifend etae Mart a S EE 89 miulsidiarelioe iliicpe TERT 91 Willst 92 Setting the Date and TM 92 Editing the Wavelength Table 5i susviss va a ue d o NO x a ER A E a n ERN 93 Specifying Data Output and Reporting Options eennnnn mnn 93 Selecting Read OPTIONS 95 Instrument QualifICatiOn 25s ruas sk Ru va WEREREREAERERENAM EE KM NR FEE REF E RED A Recommendations for Achieving Optimum Performance eeeeeeee 98 Reco
8. Available Readers Help Quant on CO ELxsS00 MB on COR EL B00 on COM EL 808 on COM EL 808 l on COM FL600 on COMZ FLe800 on COM FLe800 l on COM Port Fiters Wavelengths Current Reader Figure 28 Reader Selection dialog BioTek Instruments Inc Reading a 384 Well Plate in KC4 183 5 Click Current Reader to establish communication with the reader then click Close to return to the main screen If the test fails KC4 will provide appropriate instructions for resolving any problems 6 Atthe main screen select Data New Plate The New Data File dialog will appear New Data File X Select a Protocol E Empty Fratacal Path Cancel Browse Help Figure 29 New Data File dialog 7 Select Empty Protocol and click OK 8 Select Protocol Reading from the main screen The Reading dialog will appear 9 Define the Reading parameters gt Selecta Reading Type gt Define the Wavelength s at which the plate will be read gt Set the Plate Type to 384 WELL PLATE gt Define other reading parameters as necessary Click the Help button for assistance UQuant Operator s Manual Appendix B Using 384 Well Plate Geometry Reading Type Reader End Point ulduant on COMMS Kinetic Kinetic Stanning Spectrum Scales Calibrate Fre Headings Blank Plate AA Size 15x24 Pathlength Corre
9. BioTek Instruments Inc Liquid Testing 117 Liquid Testing Conducting Liquid Tests confirms the uQuant s ability to perform to specification with liquid samples Liquid testing differs from testing with the Absorbance Test Plate in that liquid in the wells has a meniscus whereas the test plate s neutral density glass filters do not The optics characteristics may differ in these two cases thus alerting the operator to different types of problems The liquid tests will help to detect optical defects or contamination that can contribute to errant readings Liquid Test 1 tests the alignment accuracy and repeatability of the reader and can be used for routine testing if you have the Absorbance Test Plate see page 121 Liquid Test 2 can be used to test the linearity repeatability and alignment of the reader if you do not have an Absorbance Test Plate Prepare the series of solutions of varying absorbances as described on page 123 Liquid Test 3 is an optional test offered for sites that must have proof of linearity at wavelengths lower than those attainable with the Absorbance Test Plate This test is optional because the reader has good front end linearity throughout its wavelength range see page 127 BioTek offers a dye solution PN 7120779 25 ml or 7120782 125 ml that may be used in the stock solution formulation for Liquid Tests 1 and 2 or if you prefer you may use the dye solution described in Table 4 page 119
10. 2 and 3 incorporated recommendation from Manual Update H1 to shake the plate or wait between pipetting and read steps Appendix A Added new section Controlling the Reader with Gen5 Appendix B Added new section Reading a 384 Well Plate in Gen5 BioTek Instruments Inc Intended Use Statement XV Intended Use Statement The Quant is a single channel automated bench top general purpose Enzyme Immunoassay Analyzer that performs analyses and DNA quantification of a variety of enzyme immunoassays Assay protocol variations are addressed by the developer of the ELISA test kit in accordance with the test kit s procedure A versatile curve fitting and statistical software program is preloaded on every Quant The operator can use BioTek s Extensions Define Protocol software to define up to 57 assay protocols and blanking patterns that can be stored in memory and instantly accessed 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 BioTek software package such as Gen5 KCjunior or KC4 for Microsoft Windows e Inthe European Union This product may only be used for Research and Development and other nonclinical purposes e Inall other jurisdictions This product may be used for Research and Development and in vitro Diagnostic purposes D The performance characteristics of the data reduction software
11. Dispense Shake Delay Kinetic Monitor well Seb Temperature Plate Quk In Stop Resume Synchronized Modes well Plate Plate Type EZ Description Comments Validate Cancel Help Figure 24 Procedure dialog 10 Click Read to open the Read Step dialog Select a Read Type Read Step E X Step Label default Full Plate Detection Method Absorbance Read Type Pathlength Correction m Read Speed Normal Wavelengths Figure 25 Read Step dialog BioTek Instruments Inc Le 12 13 14 L3 16 Reading a 384 Well Plate in Gen5 181 Define the Wavelength s at which the plate will be read Define other reading parameters as desired Click the Help button for assistance When complete click OK to return to the Procedure dialog Click Validate if you would like Gen5 to verify the defined read parameters If any parameters are incorrect or incomplete follow the instructions provided by Gen5 for correcting the parameters Click OK to save and close the Procedure dialog The Plate Reading dialog will appear x Date Tuesday January 03 2006 Time 09 44 46 Plate ID FO Barcode P Prompt 1 Po Prompt 4 Prompt 2 Prompt 5 Po Prompt 3 Prompt 6 Po Comments Enter Manually Cancel HEAD Read From File Simulate Read Help Figure 26 Plate Reading dialog gt Ifthe Plate
12. Figure 25 Read Step Dialog essseseeee en nnnnnmhhnnnnnmenn nnnm nnn 180 Figure 26 Plate Reading Dialog eere nnnm nenne 181 Figure 27 KC4 Main Menu System Readers eeennn nnn 182 Figure 28 Reader Selection Dialog ccseeeeeee rennen nnn 182 FEigure 29 New Data File Dial inside 183 Figure 30 Readlig DialOoG ssesesaxuteraaua cabane bardas 184 Figure 31 Setup Reader Dialog iii 185 Figure 32 Reader Setup Dialog eese nnnm nnn 185 Figure 33 Read Plate DIalOQ siii 186 Figure 34 Protocol Definition Dialog cessere 187 Figure 35 Samples With Calls on Matrix Report cccceeeeeeeeeeseeeeeeeeeeeens 190 FIgure 505 CURVE rit Rep O E usaed cds M PUN OS 191 Figure 37 Samples With Calls on Column Report eeennnnnnn nnn 192 Figure 38 Column Report Without Samples enn nnn 193 Figure 39 Panel Repo Einicio Asa XE RYE arias 194 Figure 40 Assay Detail Report Sheet 1 Of 2 cece ccc ceeeeeeeeeeeeeeeeeneeeees 195 Figure 41 Assay Detail Report Sheet 2 of 2 00occccccnccccccnncncnna anna 196 Figure 42 Spectral S6CaDissssumese nias dd CRAS 197 List of Tables Table 1 Preprogrammed Assay List eere mme 40 Table 2 Recommended Qualification Schedule enn nnne 99 Table 3 Typical Enzyme Substrate Combinations and Stopping BS OU O S CNET TT UT A m 118 Table 4 Stock Solution Formul
13. Getting Started With KCA T daa E 171 Controlling the Reader with KCjunior lessen nnnm nme 173 Sering UD RECTO Pasaia 173 migolol pne tct PE 174 Getting Started with KCjunior cassis tee ER et aa Sup a er aea c ae da iod era aie dotes 174 Using 384 Well Plate Geometry eere 177 Reading a 384 Well Plate in Gem liuius nnnm neni 178 Reading a 384 Well Plate in KC4 asas 182 Reading a 384 Well Plate in KCjunior sssssssssssssnnnnnrnrsssnsrrnenrsnnernrrrnrne 185 Report Format oeisovkREXxERE MER X RE ci dc A Cw A LOS UQuant Operator s Manual viii Preface List of Figures Figure 1 External Components of the uQuant eeennnm mnn 11 Figure 2 Removing the Top End CaP cccccccccnnncnccnncnn cnn HH 15 Figure 3 Removing the Reader and Accessories from the Outer Sd OND OX RI E CETTE UTE 16 Figure 4 Removing the Black Dress Screws from the Rear Panel 17 Figure 5 Removing the Shipping Block and Inserting the Black Dress Screws from the Rear Pariel cvsise sa vx ax vUa S PREX VER tal Y DEVE TE RA TER E EOM PUES ER 18 Figure 6 Attaching the Shipping Block and Mounting Hardware to the O Tr 19 Figure 7 Connectors for Printer Parallel Computer Serial and POW CP SUDDIV erp CS 25 Figure 8 Removing the Carrier Shipping Block and Mounting HAWAI aa 29 Figure 9 Removing the Black Dress Screws from the Carrier Shipping Block Mounting Holes and Attaching Them
14. If KC4 fails to communicate with the reader and displays a serial communications error check the cable plug in location to ensure that it matches the setup choices in Table 7 and is not a Null cable If this is suspected add another Null and try again If an Incorrect Reader Model Connected message is displayed click OK to exit the dialog and select System Readers Available Readers Verify that the reader selected is correct Getting Started with KC4 The following 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 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 Select Protocol Reading The Reading parameters dialog will appear Select a Reading Type Define the Wavelengths at which the plate will be read Select a Plate Type from the drop down list a x ae ae M Define other reading parameters as necessary Click the Help button for assistance When complete click OK 9 Select Data Read Plate The Plate Reading dialog will appear UQuant Operator s Manual 172 Appendix A Controlling the Quant with Gen5 KC4 or KCjunior Software 10
15. MAP before creating blank or control validation formulas 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 Each blank well on a plate should have an OD of less than 0 050 the formula is BLK lt 0 050 gt Each Positive Control replicate must have an OD higher than 1 000 but less than 2 500 this can be accomplished with one formula PC gt 1 OOOANDPC lt 2 500 Or with two separate formulas PC gt 1 000 and PC lt 2 500 Negative Control mean must have an OD of less than 0 100 the formula is NC x lt 0 100 BioTek Instruments Inc Define Method Map Formula and Curve 67 Number of Required Controls Blanks If a control or blank validation formula is defined enter the number of valid controls or blanks that must meet the criteria established by that formula PC E NUMBER OF VALID REPLICATES REQUIRED Use the numeric keys to enter the NUMBER OF REQUIRED CONTROLS The range is 1 through the number of defined replicates of a control or blank Press the CLEAR key to clear the displayed value Press ENTER to save the displayed value and advance to the next screen or use the Previous Screen key to move backward through the menu structure Assay Validation Formulas Assay Validation formula s establish a set of criteria used to determine whether or not an assay can be consider
16. P ACROSS and P DOWN are displayed on three screens e Select the BLANK MAP type see the descriptions on the next page e Press MORE to cycle through the available options ROW or COLUMN and P ACROSS or P DOWN e Press ENTER to save the well location and continue BioTek Instruments Inc Define Method Map Formula and Curve 53 Blank Map Definitions AIR performs an initial reading on air just prior to the plate read and uses that value as the blank value This value is subtracted from each well on the plate FULL enables a single blank well or an average of blank wells to be subtracted from the whole plate CONST Constant allows entry of a user specified absorbance value This value will be subtracted from each well on the plate Tip for using CONST Use a blank value from the first plate or a blanking plate to save space on subsequent assay plates ROW enables a single blank well or an average of blank wells to be selected for each row The maximum number of blanks is 48 The blank or average will be subtracted from each well in the row Use manual mapping to position blanks controls standards and samples COLUMN enables a single blank well or an average of blank wells to be placed in each column Since the maximum number of blanks is 12 and if all 12 columns are used each column can have only one blank Manual mapping is recommended in this case Replicates follow in the same col
17. Packing the reader and accessories BioTek Instruments Inc Repackaging and Shipping the yQuant 33 Preparing the Shipping Container 1 Place top foam on the instrument Figure 12 2 Obtain a Return Materials Authorization RMA number from BioTek s Technical Assistance Center through BioTek s Web site fax or e mail address listed in Chapter 1 3 When obtaining the RMA explain whether the reader requires calibration cleaning periodic maintenance warranty work and or repair Make a note of any error messages displayed and their frequency 4 Provide BioTek with the name and contact information of a person who may be contacted if questions arise 5 Close the box and tape it shut 6 Write RMA and the RMA number in large clear letters on the outside of the shipping container and ship the instrument to the BioTek address provided in the Product Support amp Service section of Chapter 1 Top Foam Figure 12 Preparing the shipping container UQuant Operator s Manual 34 Chapter 3 Installation BioTek Instruments Inc Chapter 4 Operation This chapter includes instructions for operating the uQuant and its onboard software jouant Front Ranes 36 OVEIVIS pe T mem 38 SY SECM ta tubis naa 38 ML MCMC acutatay mitos aia barcovrUPPPbPrsRDDE Verbi LDUDUMDLO PI PLI UDLHO DIOE Rot EU 39 Bgm E E 40 Selecting an Assay to Define eese nnnm 41 Editing
18. The purpose of the formulation is to create a solution that absorbs light at 2 000 OD full strength when dispensed at 200 ul in a flat bottom microplate well 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 that may be used as alternates to the described dye are shown in Table 3 page 118 UQuant Operator s Manual 118 Chapter 5 Instrument Qualification Repeatability This test ensures that the instrument meets its repeatability specification by reading a color absorbing solution five times without removing the microplate A dye solution that absorbs at 405 nm is acceptable but other dyes that absorb at other wavelengths may also be used Two columns of the microplate are filled with the test dye and the well with the greatest variance between readings is reviewed for compliance with the specification If the reader fails the repeatability tests review the following possible problems and solutions e Check the microwells to ensure that there is no debris that may have shifted and caused changes e Check the microplate carrier to ensure that all the holes are free of debris that may have shifted and caused changes If these remedies are ineffective call your BioTek representative or BioTek Technical Support Alignment This test confirms the mechanical
19. and accessories for signs of damage If the reader is damaged notify the carrier and your manufacturer s representative Keep the shipping cartons and packing material for the carrier s inspection The manufacturer will arrange for repair or replacement of your instrument immediately before the shipping related claim is settled Refer to the unpacking instructions and Figures 2 through 6 on the following pages when removing the instrument and its accessories from the shipping container See Repackaging and Shipping the p Quant at the end of this chapter for complete shipping instructions Packing Materials Outer shipping container 7272054 Inner shipping container accessories 7272055 Top foam end caps 7272057 Bottom foam end caps 7272056 2 Mil poly bag uQuant 98085 8 Y2 x 11 bubble bag power supply 91083 Required Tools PN Combination slotted Phillips head screwdriver 98145 BioTek Instruments Inc Unpacking and Inspecting the uQuant 15 Unpacking the Instrument and Its Accessories 1 Carefully open the top of the box 2 Remove the top foam end caps manual and Declaration of Conformity Figure 2 Declaration of Conformity Manual End Caps Figure 2 Removing the top end cap UQuant Operator s Manual 16 Chapter 3 Installation 3 Lift the reader out of the box Figure 3 and place it on a level surface Remove the reader from the poly bag Remove the accessories
20. most recent sets of plate data are stored in memory gt The form in which the results are presented 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 the assay s settings such as wavelengths numbers of well types and curve fit parameters e Select LIST to print a list of all assays name and number currently programmed in the Quant BioTek Instruments Inc Printing Reports 89 Results Report The reader stores the 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 01 23 04 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
21. 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 multiplied by 1 0 8004 0 010 equals 0 008 and when added to the 0 005 0 008 0 005 equals 0 013 which is the allowed deviation for well A1 Since the standard deviation for well A1 is less than 0 013 the well meets the test criteria Linearity Test 1 For each of the three dye concentrations calculate the mean absorbance for the wells containing that solution mean of wells A1 to H2 A3 to H4 and A5 to H6 Perform a regression analysis on the data to determine if there is adequate linearity Example using Microsoft Excel In a spreadsheet enter the three mean values in ascending order and label the column as the Y values Enter 0 50 0 75 and 1 00 and label the column as the X values Select Tools gt Data Analysis Regression Identify column Y as the Input Y Range and column X as the Input X Range and then click OK to perform the analysis If the Data Analysis command is not available on the Tools menu you may need to install the Analysis ToolPak in Microsoft Excel Consult Excel s help system for assistance Expected Results Since it is somewhat difficult to achieve high pipetting accuracy when conducting linear dilutions an R Square value greater than or equal to 0 990 is considered adequate BioTek Instruments Inc Chapter 6 Maintenance and Decontamination This chapter
22. 1 Introduction Updated descriptions of features and package contents pages 3 and 4 Clarified Specifications pages 5 and 6 Updated Technical Support page 7 Chapter 2 Instrument Description Enhanced component description page 10 BioTek Instruments Inc Revision History Continued XII Revision History Continued Revision Date G 5 04 H 11 04 Changes Chapter 3 Installation Updated all illustrations Clarified installation instructions Chapter 4 Operation Revised display screens throughout Clarified operation instructions Chapter 5 Performance Verification and IQ OQ PQ Procedures Updated entire chapter Clarified liquid test procedures Added new Chapter 6 Maintenance and Decontamination Added new Chapter 7 Troubleshooting and Error Codes Added instructions for controlling the reader with KC4 to Appendix A Computer Control formerly Appendix B Added instructions for using KC4 to manipulate data to Appendix B Using 384 Well Geometry formerly Appendix D Updated sample reports in Appendix C Report Format Removed previous Appendix A Decontamination Removed Appendix E Error Codes Removed references to General Formula pages 41 62 63 73 and 89 Preface Updated Safety Symbol text Chapter 1 Introduction Corrected Service Manual part number listed under Optional Accessories page 4 Changed VAC to V pag
23. 146 2006 10237203 AM User Administrator Company Comments System Test performed during IQ Test Results Operator ID Notes 11 53AM 01 06 06 SYSTEM SELF TEST Lambda 405 Gain 2 10 Resets 4 Channel Ref 1 Air 8656 39546 Dark 5021 5290 Delta 3635 34256 Lambda 450 Gain 1 73 Resets 4 Channel Ref 1 Air 8497 39846 Dark 5019 5241 Delta 3478 34605 Lambda 490 Gain 2 29 Resets 8 Channel Ref 1 Air 8338 39832 Dark 5018 5096 Delta 3320 34736 Lambda 550 Gain 1 74 Resets 2 Channel Ref 1 Air 8194 39947 Dark 5025 55 7 Delta 3169 34370 Lambda 630 Gain 2 51 Resets 1 Channel Ref 1 Air 7899 39176 Dark 5047 6786 Delta 2852 32390 Lambda 730 Gain 1 77 Resets 2 Channel Ref 1 Air 7552 39440 Dark 5025 5584 Delta 2527 33856 Channel Ref 1 Noise Max 5031 5380 Noise Min 5028 5377 Delta 3 3 Figure 15 Sample output for the System Test sheet 1 of 2 uQuant Operator s Manual 104 Chapter 5 Instrument Qualification AUTOCAL ANALYSIS Upper Left Corner x 9220 y 10780 Lower Left Corner x 9208 y 16312 Lower Right Corner x 532 y 16292 Upper Right Corner x 536 y 10784 Delta 1 9220 9208 12 Delta 2 536 532 4 Delta 3 10784 10780 4 Delta 4 16292 16312 20 SYSTEM TEST PASS 000 Reviewed Approved By For Technical Support In the U S In Europe BioTek Instruments Inc BioTek Instruments GmbH Tel 800 242 4685 Tel 49 0 7136 9680 Fax 802 655 33
24. 580 590 600 Lambda Figure 42 Spectral Scan UQuant Operator s Manual 198 Appendix C Report Format BioTek Instruments Inc
25. Assay Detail Report Sheet 2 of 2 cccccccccsscueueeeeuesuuuueeueuenauuesauuess 196 Spectral SCAN creci 197 190 Appendix C Report Format Bio Tek Instruments Assay Open Assay 04 Date 03 16 04 Lot Time 01 05 34PM Operator Wavelength 405 Temp Plate ID COMMENTS E 2 3 4 5 6 7 E 9 10 11 12 A NEG POS 1 501 0 112 SMP31 SMP39 CALL VAL CalcOD 0 109 Well RE RSLT B CALL VAL CalcOD 0 119 Well PC RSLT C CALL POS Calc0D 0 290 Well SMP1 RSLT D CALL POS Calc0D 0 301 Well SMP2 RSLT E CALL Calc0D Well RSLT NEG 1 672 SMP32 NEG 2 291 SMP33 NEG NEG 2 643 2 032 SMP34 SMP42 F CALL CalcOD Well RSLT G CALL CalcOD Well RSLT H CALL CalcOD Well RSLT Figure 35 Samples with calls on Matrix Report BioTek Instruments Inc 191 Bio Tek Instruments Assay ASSAY 01 Date 03 16 04 Lot Time 02 27 57PM Operator Wavelength 450 Temp Plate ID COMMENTS Standard Curve Linear Fit 0 310 y err C5 m E CX0 LOS CO co eo X c ho n d Poo CONCENTRATION Curve y Ax B Coefficients A 0 035 B 132 R Sqr 1 0000 Figure 36 Curve Fit Report Quant Operator s Manual 192 Appendix C Report Format Bio Tek Instruments Assay Open Assay 04 Date 03 16 04 Lot Time 01 05 34PM Operator Wavelength 405 Temp Plate ID COMMENTS Interpretation of Results GREY
26. Contents iia noia 4 UQuant Operator s Manual IV Preface Optional ACCESO NOS ia iii dodi Cord 4 SPECITICAUIONS uns rien 5 MICKODIGUOS anar 5 SPCC Ol REAGING nase iwahatrasan aaa aa aa aaa 5 Optical Specifications susi es ees he ee ea laa eee ae 5 Hardware and Environmental Specifications ccccccceseueeesuecenaueeseuevenaues 6 Product SUDDOrt amp SEVICE essc exe bU ade tavi doe rcr e a i dove vet ete D anes 7 Contacting the Technical Assistance Center TAC cccccseeeeeeeeeeeeeteneens 7 Instrument Descrip ION ini PRINCIDIES OF ODerati N sonia ais 10 External COMpoOnentsaVini didas 10 ERASMUS Unpacking and Inspecting the Quant cc eeccecsee eee eeeeeeeeeeeeeeeteneeeeteageees 14 Unpacking the Instrument and Its Accessories ccccceeeeeeeeeeeeeeeeeeeeeeneees 15 Sebring pne OB scan adi 20 Operating ENVIO Men lalo 20 Electrical Connection saaana aiii 20 Power Up and System Test occccccconnccancnnn na rca raro rn aan nne 21 HOUanC Malit Mena oo 21 Adjusting the Wavelength Table eese ennmnnnnnmnhn menm nnn 22 Connecting a Printer to the uQUaMt cccccccccccncccncnnnnnnn nennen nnn 24 Optional Setting Up the Serial Port for Communications eeesee 26 Attaching tege csscdeebesieatspbe bstva deste shoe Da a xaO URS A peat 26 Installing the PC SON Wars 26 Setting Communication Parameters csceseee enean nma nra 26 Repackaging and Shipping the puQuant
27. Enter any desired information place the plate on the carrier then click START READING to begin the plate read gt The plate will be read and then the raw data results will display in KC4 gt To analyze manipulate or print results protocol parameters should be defined Refer to KC4 s Help system or User s Guide for instructions BioTek Instruments Inc Controlling the Reader with KCjunior 173 Controlling the Reader with KCjunior Before installing KCjunior software verify that your computer meets the minimum system requirements specified in KCjunior s User s Guide or Help system Setting Up KCjunior The yQuant M can be operated using a computer running BioTek s KCjunior software The following instructions briefly show you how to set up KCjunior for operation of the reader Refer to KCjunior s User s Guide or Help system for more detailed instructions 1 Turn off the computer and reader Connect the appropriate serial cable PN 75053 between the two machines See Table 7 on page 175 for the cable pinouts 2 Turn on both machines 3 Install KCjunior on the computer s hard drive and register the software with BioTek 4 Once installed start KCjunior Select Setup then Reader1 or Reader 2 to open the Reader Setup dialog 6 Scroll through the Reader Type list and select the uQuant Define the communication parameters as follows gt Com Port COMn select the COM serial port used for
28. Inc Define Method Map Formula and Curve 63 Formula Type The uQuant supports three types of formulas as well as the ability to define variables for use within Transformation formulas Note GENERAL formulas are not used LECT FORMULA TYPE T OFF T RAN S VAL LECT FORMULA TYPE NERAL TRAN S VA R 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 raw 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 The TRANS VAR option allows you to define a variable to be used in transformation formulas UQuant Operator s Manual 64 Chapter 4 Operation Formula Entry After the formula type is selected the FORMULA ENTRY screen appears Each formula can contain a maximum of 24 characters Spaces are not necessary Note In formulas OD is used to represent the optical density value FORMULA MAT H O T e After a moment the FORMULA 1 prompt disappears and the formula can be entered Use the options found under MATH OTHER MAP and FUNCTN to build the formula
29. Instruments Inc Reading a 384 Well Plate in KCjunior 187 Protocol Definition Endpoint El El l le le m P EL T lice etna ber eity INCUbStiaA Murator lemperature E e LS GrTHIFIBIGIUIS stare rea Mode Figure 34 Protocol Definition dialog 8 Place the 384 well plate on the plate carrier then click OK to read the plate Click OK when prompted to Place the plate on the carrier gt The plate will be read and then the raw data results will be displayed in KCjunior Toanalyze manipulate or print the results Protocol parameters should be defined Refer to KCjunior s Help system or User s Guide for instructions Quant Operator s Manual 188 Appendix B Using 384 Well Plate Geometry BioTek Instruments Inc Appendix C Report Format This appendix contains examples of reports that can be generated and or printed from the uQuant Refer to Printing Reports in Chapter 4 for details on how to print these reports In addition an Assay List Assay Definition Map and Result can be printed by choosing Report from the Main Menu screen Samples With Calls on Matrix Report eenn nmn 190 Curve FIC RepoO rusia dais ci 191 Samples With Calls on Column Report cccccccccnnncccccnnncnnc cnn 192 Column Report Without Samples eseen nnnm 193 Panel Reporta TC HT 194 Assay Detail Report Sheet 1 Of 2 0 00ccccccnccccncccnnncana nr rn rns 195
30. Map Report for each assay that will be included in the Panel To create a panel assay start at the Main Menu select DEFINE and then choose assay number 99 Enter the panel assay name e The default name is PANEL e Use the alpha and numeric keys to update the assay name if desired e Press ENTER to continue The NUMBER OF ASSAYS screen will appear BioTek Instruments Inc Define Method Map Formula and Curve 79 Specify the number of assays to include in the panel 1 to 8 Press ENTER to continue The MAPPING DIRECTION selection screen will appear P PING DIRECTION DOWN W N ACROSS This option ensures that all assays will be mapped in the same direction Select DOWN or ACROSS D The original mapping directions for the predefined assays are overwritten by the Panel s mapping directions If the assay includes replicates they will follow the Panel mapping direction After selecting the mapping direction of the assays choose which assays to include in the panel LECT AS SAY NUMBER ME HBS AG 1 e Press Options 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 an assay is selected its starting location must be defined uQuant Operator s Manual 80 Chapter 4 Operation START MAPPING A T WELL LOCATION Use the alpha and numeric keys to choose the well location to begi
31. alignment of the instrument by comparing the measurements read with a test solution in the microplate with those obtained when the plate is rotated so that A1 is now in the H12 position The first dye OD readings from the repeatability testing listed above are compared to the same readings when the plate is turned 180 This indicates if the same well will give similar values in the opposite position on the carrier If the readings do not meet the specifications provided in Chapter 1 the reader may be out of alignment Table 3 Typical Enzyme Substrate Combinations and Stopping Solutions Substrate Stopping Solution Alkaline o nitrophenyl phosphate 3N sodium hydroxide Phosphate beta o nitrophenyl beta D galactopyranoside 1M sodium carbonate Galactosidase Peroxidase 2 2 Azino di ethylbenzothiazoline sulfonic citrate phosphate acid ABTS buffer pH 2 8 o phenylenediamine 0 03N sulfuric acid BioTek Instruments Inc Liquid Testing 119 Stock Solution Formulation The stock solution for Liquid Tests No 1 and No 2 may be formulated from the chemicals listed below Solution A or by diluting a dye solution available from BioTek Solution B Solution A Required Materials e Deionized DI water e FD amp C Yellow No 5 dye powder typically 90 pure e Tween 20 polyoxyethylene 20 sorbitan monolaurate or BioTek wetting agent PN 7773002 e Precision balance with readability of 0 001 g e Weigh boat e 1 lit
32. cesses nnne nennen nnn 168 Controlling the Reader with KC4 eise nnnm nme 170 SENT UP A Opr CEDE 170 PFODIOITIS esas ae osrebbaid aps vore odd Er sd 171 Getting Started With KA c ooseshutsatue een a ode acea da i ear et a eit vx 171 Controlling the Reader with KCJUNIO ssssssesseeeeenn nmm 173 oetung UP KC UNION senior variar 173 PROD MN SO 174 Getting Started with KOJUNIOF occcoccccccccccccnnnarrcrrcnr rre 174 166 Appendix A Controlling the Quant with Gen5 KC4 or KCjunior Software Overview BioTek s Gen57M KC4 or KCjunior software provide the user even more power and flexibility For example the uQuant M 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 This Appendix provides instructions for controlling the Quant with BioTek s Gen5 KC4 or KCjunior software BioTek Instruments Inc Controlling the Reader with Gen5 167 Controlling the Reader with Gen5 Before installing Gen5 software verify that your computer meets the minimum system requirements specified in Gen5 s Help system or Getting Started Guide Setting Up Gen5 The wQuant can be operated using a computer running BioTek s Gen5 software The following instructions briefly show you how to set up Gens for operation of
33. contains procedures for maintaining and decontaminating the uQuant Maintenon C siue EI uL seca eee oes I LL PE RU E P M ADITU IE 132 Routine Cleaning Procedure eeesseeeeeee ennemi 132 Decontamination PFOCOGOUFG scito A o LO 134 132 Chapter 6 Maintenance and Decontamination Maintenance A daily maintenance schedule is the best way to ensure accurate performance and a long life for your instrument Frequent cleaning of the microplate carrier and all exposed surfaces of the instrument will help to reduce the amount of particulates or dust that can cause erroneous readings Routine Cleaning Procedure A A Purpose Warning Internal Voltage Always turn off and disconnect the reader from the power supply for all cleaning operations Do not immerse the instrument spray it with liquid or use a wet cloth Do not allow the cleaning solution to run into the interior of the instrument If this happens contact the BioTek Service Department Do not soak the keypad this will cause damage Moisten a clean cloth with deionized or distilled water and wipe the keypad Dry immediately with a clean dry cloth Do not apply lubricants to the microplate carrier or carrier track Lubrication on the carrier mechanism or components in the carrier compartment will attract dust and other particles which may obstruct the carrier path and cause the instrument to produce an error Exposed surfaces may be cleaned not deco
34. criteria below The last number is the lambda table position number This error indicates one of the following situations e The measurement channel failed lt 100 during optic test with the flash on The measurement channel failed lt 100 during a read or blank read not in sweep mode with the flash off The measurement channel failed lt 100 or if the Dark value has changed more than 10 from the last self test data during a read or blank read with the flash on UQuant Operator s Manual 156 Chapter 7 Troubleshooting and Error Codes General Errors Continued Probable Causes Absorbance analog PCB or measurement channel analog PCB is defective Shielding of the cable between measurement channel and analog PCB is defective or disconnected Measurement channel photodetector is defective or the optic spray is damaged A faulty analog PCB or faulty internal grounding may cause internal electronic noise There may be an ambient light leak Ensure that the plate carrier door is properly closed Electrical noise may be penetrating the measurement chamber The bottom and top shrouds are part of the electrical shielding Verify that the shrouds are installed and properly fastened Order sorting filter wheel is jammed and not aligning the filter wheel to block the light path or the filter is degraded and not passing Reference Channel Air Blank out of range See criteria below This error is indicating the Air
35. e Press ENTER to advance to the EDIT ASSAY NAME screen You may change the default assay name to a more descriptive one see Editing the Assay Name on the next page CLEAR Clears the reader s display gt 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 UQuant Operator s Manual 42 Chapter 4 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 e The cursor is positioned at the first editable field e g under H 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 Z e Use the left 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 the CLEAR key to remove the assay name from the display e Use SOFT KEYS 1 2 3 and 4 to select a dash forward slash colon or space for inclusion in the assay name BioTek Instruments Inc Define Method Map Formula and Curve 43 Define Method Map Formula and Curve The DEFINE screen allows you to edit the Method Map Formula or Curve Fit parameters for
36. for instructions BioTek Instruments Inc Define 41 3 Optional Edit the assay name then press ENTER See Editing the Assay Name on the next page for detailed instructions The DEFINE menu will appear DEFINE METHOD MA P FORMULA CURVE The following options are available within the DEFINE menu e METHOD Define the wavelength type single or dual wavelength s and plate geometry page 43 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 page 47 e FORMULA Define cutoff transformation and or validation formulas Create variables to be used within formulas page 62 e CURVE Specify a curve fit type and x y axis types lin log Specify whether or not standard outliers can be edited and then the method by which they will be edited Enable or disable the extrapolation feature page 74 Selecting an Assay to Define To select an assay to define or modify start at the Main Menu and select DEFINE to display the SELECT ASSAY NUMBER screen SELECT ASSAY NUMBER NAME H B S AG 1 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
37. have not been established with any laboratory diagnostic assay The operator must evaluate this software in conjunction with the specific laboratory diagnostic assay This reevaluation must include the establishment of new performance characteristics for the specific assay 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 Specimen Preparation Samples should be obtained treated and stored following instructions and recommendations contained in the package kit UQuant Operator s Manual XVI Preface Repackaging and Shipping D If you need to ship the instrument to BioTek for service or repair contact BioTek for a Return Materials Authorization RMA number and be sure to use the original packing Other forms of commercially available packing are not recommended and can void the warranty If the original packing materials have been damaged or lost contact BioTek for replacement packing Warnings Operate the instrument on a flat surface and away from excessive N humidity Bright sunlight or strong incandescent light can reduce the linear performance range of the instrument Measurement values may be affected by extraneous particles such as dust in the microplate
38. hot surface Attention surface chaude Warnen heifse Oberflache Precauci n superficie caliente Attenzione superficie calda Consult instructions for use Consulter la notice d emploi Gebrauchsanweisung beachten Consultar las instrucciones de uso Consultare le istruzioni per uso In vitro diagnostic medical device Dispositif m dical de diagnostic in vitro Medizinisches In Vitro Diagnostikum Dispositivo m dico de diagn stico in vitro Dispositivo medico diagnostico in vitro Caution refer to accompanying documents Attention voir documents d accompanement Achtung siehe Begleitpapiere Atenci n vease los documentos incluidos Attenzione consultare la doc annessa Separate collection for electrical and electronic equipment Les quipements lectriques et lectroniques font l objet d une collecte s lective Getrennte Sammlung von Elektro und Elektronikger ten Recogida selectiva de aparatos el ctricos y electr nicos Raccolta separata delle apparecchiature elettriche ed elettroniche BioTek Instruments Inc Chapter 1 Introduction This chapter introduces the uQuant and describes its hardware and software features Also included are information about this manual and instructions for obtaining technical assistance if needed Introducing INE DOUSPE ss x e Oo laa t Cet Era o ud ace e t ce As OU alley Condos robado qM SED PLE DI Hardware Features 2s buta Aa atc adici aq Ao pom CA GE td cena oes SoftWare Feature
39. if EDIT STANDARD OUTLIERS was set to MANUAL in the assay definition the option to edit outliers is presented 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 If AVERAGE STANDARDS was set to NO in the assay definition one or more standard replicates can be chosen for exclusion UQuant Operator s Manual 90 Chapter 4 Operation ED IT STD 1 RE P 1 Y E O e Select YES to exclude the replicate from curve fit calculations e Select NO to retain the replicate e Press ENTER to advance to the next replicate gt f AVERAGE STANDARDS was set to YES in the assay definition one or more standard groups can be chosen for exclusion e Select YES to exclude the group from curve fit calculations e Select NO to retain the group e Press ENTER to advance to the next group D 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 fail BioTek Instruments Inc Printing Reports 91 Printing Results After the assay is selected and standard outliers are edited if necessary the results report can be printed PRINT
40. is the difference between the two readings the delta OD Dual wavelength readings can significantly reduce optical interference caused by scratched or fingerprinted microplates since the scratches or fingerprints reduce the amount of light on both wavelengths BioTek Instruments Inc Define Method Map Formula and Curve 45 Meas Selection The MEAS selection screen allows you to select the wavelength s for the assay Press MORE to cycle through the available options The six wavelength values presented here for selection come from the uQuant s wavelength table You can change the contents of the table by selecting UTIL gt SETUP LAMBDA from the Main Menu See Editing the Wavelength Table on page 93 for more information e Select the MEAS primary wavelength e Use the right arrow key to move the cursor to REF and then select the Reference Wavelength e Press ENTER to continue Plate Type The PLATE TYPE selection screen allows you to select the geometry of the plate that will be used for the assay Press MORE to cycle through the available options The pQuant does allow you to select a Plate Type of 384 however it does not print results reports for assays with this plate type After reading a 384 well plate the Quant sends the raw data to a host PC where it can then be printed and or further analyzed using data analysis software The most effective and versatile method for reading 384 well plates is
41. memory is corrupt Replace the processor PCB e If the error persists contact BioTek TAC UQuant Operator s Manual 162 Chapter 7 Troubleshooting and Error Codes General Errors Continued A D calibration STBY line never went low A D calibration STBY line went low but never transitioned to a high The analog PCB circuit failed when trying to initialize the A D or the cable to the defective PCB has lost continuity Probable Causes e Problems with the absorbance analog PCB or the cable between the analog PCB and motor power supply PCB Invalid parameter value selected This error can occur only during computer control indicating that an invalid assay configuration was sent to the instrument lt Motor gt currently in use This error indicates that the lt motor gt is not available for this model or already has a task assigned to it At the beginning of the motor_setup function the basecode checks to see if the motor is currently in use or not available This error can occur with any motor request See table below for error 2800 2801 2802 Order sorting filter wheel 2803 Probable Causes e User selected the incorrect model in computer software e Downloaded incorrect basecode Results data being sent not acknowledged by host PC This error is indicating the handshaking between the host PC software and the reader did not complete This is a lost data condition e Software development only Plate read t
42. of Gen5 s Help system 10 Click OK again to save and close the Procedure dialog The Plate Reading dialog will appear 11 Enter any desired information place the plate on the carrier then click READ to begin the plate read Click OK when the Load Plate dialog appears The plate will be read gt To view the raw data results use the Data drop down arrow in the Plate screen to select one wavelength The results will be displayed for the selected wavelength Repeat for other wavelengths gt To analyze manipulate or print results Protocol parameters should be defined Refer to Gen5 s Help system for instructions Gen5 Reader Control does not support data reduction UQuant Operator s Manual 170 Appendix A Controlling the Quant with Gen5 KC4 or KCjunior Software Controlling the Reader with KC4 Before installing KC4 software verify that your computer meets the minimum system requirements specified in KC4 s User s Guide or Help system Setting Up KC4 The yQuant M can be operated using a computer running BioTek s KC4 software The following instructions briefly show you how to set up KC4 for operation of the reader Refer to KC4 s User s Guide or Help system for more detailed instructions La 8 Turn off the computer and the reader Connect the appropriate serial cable PN 75053 between the two machines See Table 7 on page 175 for the cable pinouts Turn on both mach
43. of the self test Possible Causes e Memory corruption e New main PCB installed BioTek Instruments Inc Error Codes 161 General Errors Continued Autocalibration data missing for absorbance reads No autocal data exists for the read location Probable Cause The 7270400 PCB was changed and the Flash memory does not have the calibration values loaded Perform the Autocalibration procedure to correct this lt Motor gt not homed successfully This error indicates that the lt motor gt is not at home At the beginning of the motor_position function the basecode verifies that the motor is homed If not it sends an error and exits the function Probable Causes If an error 0200 is ignored See the probable causes for 0200 1300 0200 1301 0201 1302 Order sorting filter wheel 0202 Computer control assay definition error Used only for software development purposes 1700 Kinetic interval not correct for selected options This error indicates one of the following situations e The kinetic interval in the current assay is too short e The kinetic interval for an absorbance read the total time or kinetic interval 0 Probable Causes e User programming error Increase or decrease the kinetic interval Memory allocation failed Indicates that the process failed while saving or moving data If this occurs turn off the unit wait 30 seconds and then turn on the unit Probable Causes e The
44. power supply PCB sent the incorrect voltage request to the lamp power supply UQuant Operator s Manual 150 Chapter 7 Troubleshooting and Error Codes General Errors Continued Absorbance reference channel noise test at max gain See noise Delta on self test fail if lt 20 Significant variations in background electronic noise were detected when blocking the light and increasing the gain to maximum Probable Causes Electrical noise may be penetrating the measurement chamber The bottom and top shrouds are part of the electrical shielding Verify that the shrouds are installed and properly fastened The coaxial cable ground between the reference channel and the absorbance analog PCB may be floating or not connected There may be an ambient light leak Ensure that the plate carrier door is properly closed Analog PCB failure the photodetector is noisy A faulty analog PCB or faulty internal grounding may cause internal electronic noise Absorbance measurement channel noise test at max gain See noise Delta on self test fail if lt 20 Significant variations in background electronic noise were detected when blocking the light and increasing the gain to maximum Probable Causes Electrical noise may be penetrating the measurement chamber The bottom and top shrouds are part of the electrical shielding Check to see if the shrouds are installed and are properly fastened The coaxial cable ground bet
45. reading at the time of the plate read was lt 50 of the air reading at the time of the optic test The last number is the lambda table position number Probable Causes Flash lamp has missed flashes or erratic flash during the blank read Dirty optics or spilled substance on the optics Order sorting filter wheel is jammed and not aligning the filter wheel to block the light path or the filter is degraded and not passing or is blocking the light BioTek Instruments Inc Error Codes 157 General Errors Continued 0A11 0A16 0DOO ODO1 ODO6 Measurement Channel Air Blank out of range See criteria below The Air reading at the time of the plate read was lt 50 of the air reading at the time of the optic test The last number is the lambda table position number Probable Causes Flash lamp has missed flashes or erratic flash during the blank read Dirty optics or spilled substance on the optics The order sorting filter wheel is jammed and not aligning the correct filter to line up with the light path the filter is degraded and not hli or the filter is blocking the light Printer timed out The time allotted for the instrument to make a valid connection to a printer has expired Probable Causes e Printer not connected on line or powered up e Printer parallel port may not be correctly selected Wavelength calibration data missing during a spectral scan Wavelength data is missing prior to a spe
46. s Manual 148 Chapter 7 Troubleshooting and Error Codes General Errors Continued Reference channel light beam saturated too much light Air reading reached 65535 This error indicates one of the following situations e During an absorbance filter calibration or wavelength scan the measurement channel was saturated when storing to memory During a spectral scan read the measurement channel was saturated for one of the wavelengths Probable Causes Filter Calibration The monochromator mirror grating is damaged Absorbance analog PCB intermittently failed Missed flashes or erratic flash lamp The order sorting filter wheel is jammed and not aligning the correct filter to line up with the light path or the through hole is allowing white light to pass Voltage to the lamp has increased because the lamp power supply has failed or the motor power supply PCB sent the incorrect voltage request to the lamp power supply Spectral Scan Read e The monochromator mirror grating is damaged Analog PCB intermittently failed Missed flashes or erratic flash lamp The order sorting filter wheel is jammed and not aligning the correct filter to line up with the light path or the through hole is allowing white light to pass Voltage to the lamp has increased because the lamp power supply has failed or the motor power supply PCB sent the incorrect voltage request to the lamp power supply BioTek Instruments Inc E
47. scan the reference channel was saturated when storing to memory During a spectral scan read the reference channel reached saturation at one or more of the selected wavelengths Probable Causes Spectral Scan Monochromator has a defect in the mirror gradients Analog PCB intermittently failed Order sorting filter wheel is jammed not aligning the correct filter to line up with the light path or the through hole is allowing white light to pass Voltage to the lamp has increased because the lamp power supply has failed or the motor power supply PCB sent the incorrect voltage request to the lamp power supply Light beam saturated too much light Air measurement channel reading reached 65535 This error indicates one of the following situations e During a spectral scan read the measurement channel reached saturation at one or more of the selected wavelengths e Filter one has saturated the measurement channel Probable Causes Spectral Scan Read e The monochromator mirror grating is damaged e Analog PCB intermittently failed The order sorting filter wheel is jammed and not aligning the correct filter to line up with the light path or the through hole is allowing white light to pass Missed flashes or erratic flash lamp Voltage to the lamp has increased because the lamp power supply has failed or the motor power supply PCB sent the incorrect voltage request to the lamp power supply BioTek Instrum
48. screen initially opened in 96 well layout see Figure 23 on page 179 the screen will change to 384 well layout Enter any desired information place a 384 well plate on the carrier and then click READ to begin the plate read Click OK when the Load Plate dialog appears The plate will be read gt To view the raw data results use the Data drop down arrow in the Plate screen to select one wavelength The results will be displayed for the selected wavelength Repeat for other wavelengths gt To analyze manipulate or print results Protocol parameters need to be defined Refer to Gen5 s Help system for instructions Gen5 Reader Control does not support data reduction Quant Operator s Manual 182 Appendix B Using 384 Well Plate Geometry Reading a 384 Well Plate in KC4 1 Start KC4 Log in if prompted 2 At KC4 s main screen choose System Readers TG NewPlate created from New Test Protocol prt Data Edit Protocol Display System Diagnostics Security LogIn LogOut Audit Trail Maintain Files Registration Setup Toolbar Plate Formats Reader Control System Readers Synergy HTTR w Injectors on COM1 System Adr 4 Figure 27 KC4 main menu System Readers 3 The Reader Selection dialog will appear Select the Quant 4 Click the Port button and subsequent Setup button to define the communications parameters Reader Selection Current Reader Ouan on COM 2 Close
49. the Main Menu select UTIL The SELECT UTILITY OPTION screen will appear ELECT UT IL I T Y OPTION ESTS SETUP OUTPUT READ 3 From the SELECT UTILITY OPTION screen select TESTS The SELECT TEST screen will appear SELECT TEST SYSTEM C HK SUM CALPLA T E lt Note The desired wavelengths must be entered into the wavelength table before running the test 4 Select SYSTEM to run the System Test UQuant Operator s Manual 102 Chapter 5 Instrument Qualification To run the System Test using Gen5 select System Diagnostics Run System Test If the Instrument Selection dialog appears Gen5 and Gen5 Secure only select uQuant then click OK To run the System Test using KC4 open the System menu and select Diagnostics Run Optics Test Enter the Reader Serial Number then click Start To run the System Test using KCjunior open the Utilities menu and select Diagnostics Reader System Test Enter the Reader Serial Number then click Run Test The System Test report lists results in a Pass Fail format Figures 15 and 16 and can be viewed saved and printed Print the report to document periodic testing or troubleshoot BioTek Instruments Inc Qualification Procedures 103 Note This report varies slightly depending upon how it was run via the keypad Gen5 KC4 or KCjunior Genb System Test Report Reader uQuant Serial Number 128755 Basecode P N 7270201 v2 02 Date and Time
50. the reader Refer to Gen5 s Getting Started Guide or Help system for more detailed instructions 1 Turn off the computer and the reader Connect the appropriate serial cable PN 75053 between the two machines See Table 7 on page 175 for the cable pinouts 2 Turn on both machines 3 Install Gen5 on the computer s hard drive and register the software with BioTek gt f you purchased Gen5 s Reader Diagnostics Utility required for the Absorbance Test Plate install this software on the computer s hard drive 4 Open Geno Login if prompted Gen5 Secure The default Administrator password is admin 6 Atthe Welcome to Gen screen select System Menu 7 From Gen5 s main screen select System Reader Configuration to open the Reader Configuration dialog 8 Click the Add button to open the Reader Settings dialog gt Gen5 and Gen Secure Up to two readers may be added in Gen5 9 Use the drop down list in Reader Type to select uQuant 10 Enter the appropriate Com Port gt The Baud Rate is set at 9600 and cannot be changed 11 Click Test Comm Gen5 will attempt to communicate with the reader gt If you receive The reader is communicating message click OK and then OK again to save the settings Click Close at the Reader Configuration dialog to return to the main screen UQuant Operator s Manual 168 Appendix A Controlling the Quant with Gen5 KC4 or KCjunior Software Pro
51. to 8 results that can be stored in memory e Onboard spectral scanning e Support for the following languages English French German Italian and Spanish Contact BioTek for information on changing to a different language UQuant Operator s Manual 4 Chapter 1 Introduction Package Contents e wQuant TM Microplate Spectrophotometer e Power cord part number varies according to country of use e 24 VDC power supply PN 61062 e Operator s manual PN 7271000 e Printer cable PN 71072 e Dust cover PN 7332040 e RS 232 serial cable PN 75053 e Assay Reference Guide PN 7271006 e Unpacking instructions PN 7271003 packing instructions PN 7271012 e Shipping document PN 94075 that includes a Warranty Statement and Certificate of Compliance and Calibration e Set of shipping materials PN 7273000 e Declaration of Conformity PN 7271004 Optional Accessories e Product Qualification Package PN 7270514 e Biolek QC Check Solution 1 for liquid testing PN 7120779 for 25 ml PN 7120782 for 125 ml e BioTek wetting agent solution for liquid testing PN 7773002 e HP DeskJet printer 110 V only PN 97152 e Adapter for 60 72 and 96 well Terasaki plates PN 7330531 e Gen5 KC4 or KCjunior software PNs and versions listed on Biotek com or contact your local dealer e Gen5 Reader Diagnostics Utility PN 5320201 e Service Manual PN 7331005 e Absorbance Test Plate PN 7260522 or 9000547
52. 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 pu A cy ae 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 Define the Wavelength s at which the plate will be read Select a Plate Geometry from the drop down list 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 gt The plate will be read and then the raw data results will display in KCjunior gt To analyze or manipulate results a Protocol should be defined Refer to KCjunior s Help system or User s Guide for instructions BioTek Instruments Inc Controlling the Reader with KCjunior 175 Table 7 Serial Cable Pinout Description Serial Cable Pinout PC 9 pin female Reader 25 pin female Shell shielding For a 25 pin PC connection using BioTek serial cable PN 75053 plus 9pM 25pF adapter PN 49755 UQuant Operator s Manual 176 Appendix A Controlling the Quant with Gen5 KC4 or KCjunior Softwa
53. to the Rear Panel 30 Figure 10 Attaching the Black Dress Screws to the Rear Panel 31 Figure 11 Packing the Reader and Accessories cccceeeeeeee seen eeeeeeeeeeees 32 Figure 12 Preparing the Shipping Container eee 33 Figure 13 Quant Front Panel With 2 Line x 24 Character LCD and KEY DAG A A 36 Figure 14 Options Available from the Main Menu eere 39 Figure 15 Sample Output for the System Test Sheet 1 of 2 103 Figure 16 Sample Output for the System Test Autocal Analysis Sheeob 201 2 restet quce Eo DRE a N oq ed UV DEUM vae UM pia pt d RE 104 Figure 17 Sample Standards Certificate for the Absorbance Test Plate 109 Figure 18 Sample Absorbance Plate Test Report eene 113 Figure 19 Sample Peak Wavelength Certificate Showing Wavelength of Peak in the Interval Between 580 and 590 nm esee 116 Figure 20 Reader Configuration DialO0Q cccccccnccccccccnnncncnnc nan nnn 178 Figure 21 Reader Settings Dialog eeeeseeeeeee nennen nenne 178 Figure 22 New Experiment Dialog ccccccceeee cece eeeeee nnn 179 Figure 23 Experiment Workspace cccccecseceuecseecrensuucrensuccvensuuuuensugerenes 179 Figure 24 Procedure Dialog cese eene nnne nmnmennn nnnm nnn 180 BioTek Instruments Inc List of Figures Continued Ix List of Figures Continued
54. uQuant reader may come with preprogrammed assays already installed Refer to Table 1 under Define on page 40 for a list of preprogrammed assays System Startup To turn on the pQuant press the on off switch on the right side of the reader s base The i Quant will perform a System Test displaying the screens shown below until initialization is complete During this period all keys are inactive If the instrument fails the System Test a chirp will sound and an error code will display e Refer to System Self Test and Checksum Test in Chapter 5 for more information e Refer to Chapter 7 Troubleshooting and Error Codes to interpret error codes and Chapter 1 Introduction for information on contacting BioTek Instruments Technical Assistance Center TAC BioTek Instruments Inc Main Menu 39 Main Menu Following successful power up of the Quant the Main Menu appears 01 22PM 01 23 04 DEF INE REPORT U TI L The Main Menu permits access to all reader functions Figure 14 e READ Choose a predefined assay for plate reading Alternatively press the key labeled READ on the keypad page 83 e DEFINE Create a new assay or modify an existing one Definable parameters include assay type wavelengths well identifiers plate mapping options formulas and curve fitting parameters page 40 e REPORT Print a Result Map Assay or List Report page 88 e UTIL Run a System Test page 101 or Checksum Test page 1
55. value of 590 nm then the reader meets specifications If the reader reports 591 nm then the reader does not meet specifications If the reader fails the wavelength accuracy test review the following possible problems and solutions gt Make sure that the test plate actually has a filter in location C6 BioTek s Absorbance 6 Filter Test Plate PN 9000547 does not have this filter gt Check the C6 filter to ensure that it is clean If necessary clean it with lens paper UQuant Operator s Manual 116 Chapter 5 Instrument Qualification IMPORTANT Do not remove the filter from the test plate and do not use alcohol or other cleaning agents gt Make sure that the information entered via the keypad Gen5 KC4 or KCjunior matches the information on the Test Plate s Peak Wavelength Certificate Make sure the Test Plate is within its calibration certification period gt Check the microplate carrier to ensure that it is clear of debris If these remedies are ineffective or if the test plate requires recalibration call your BioTek representative or BioTek Technical Support MULTIBAND CALIBRATION FILTER IN LOCATION C 200 300 400 500 600 700 800 WAVELENGTH nm TYPICAL ABSORBANCE SPECTRUM DIDIMIUM GLASS V 10 WAVELENGTH OF PEAK IN THE INTERVAL BETWEEN 580nm AND 590nm Figure 19 Sample Peak Wavelength Certificate Showing Wavelength of Peak in the Interval Between 580 and 590 nm
56. wells A clean work area is necessary to ensure accurate readings When operated in a safe environment according to the instructions in this document there are no known hazards associated with the uQuant M However the operator should be aware of certain situations that could result in serious injury these may vary depending on the instrument model BioTek Instruments Inc Hazards and Precautions XVII Hazards and Precautions Hazards Warning Power Rating The instrument s power supply must be A 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 Electrical Grounding Never use a two prong plug adapter to connect primary power to the external power supply Use of a two prong adapter disconnects the utility ground creating a severe shock hazard Always connect the power cord directly to a three prong receptacle with a functional ground Warning Internal Voltage Always turn off the power switch and unplug the power cord before cleaning the outer surface of the instrument Warning Potential Biohazards Some assays or specimens may pose a biohazard Adequate safety precautions should be taken as outlined in the assay s package insert Always wear safety glasses and appropriate protective equipment such as chemically resistant rubber gloves and apron Warning Liqu
57. you need to send an instrument to BioTek for service or repair please contact our Technical Assistance Center Contacting the Technical Assistance Center TAC Our Technical Assistance Center is open from 8 30 AM to 5 30 PM EST Monday through Friday excluding standard U S holidays You can send a fax or an e mail any time Phone 800 242 4685 in the U S or 802 655 4740 outside the U S Fax 802 655 3399 E Mail tac biotek com Please be prepared to provide the following information e Your name and company information e A daytime phone or fax number and or an e mail address e The product name model and serial number e The software part number and basecode version available via the keyboard by selecting UTIL gt TESTS gt CHKSUM e For troubleshooting assistance or instruments needing repair the specific steps that produce your problem and any error codes displayed see also Chapter 7 Troubleshooting and Error Codes Returning Instruments for Service Repair If you need to return an instrument to BioTek for service or repair please contact the TAC for a Return Materials Authorization RMA number before shipping the instrument Repackage the instrument properly see Chapter 3 Installation write the RMA number on the shipping box and ship to this address BioTek Instruments Inc Technical Assistance Center 100 Tigan Street Highland Park Winooski Vermont 05404 USA UQuant Operator s Manual 8 Chapte
58. 01 23 04 D DMM Y Y gt Enter the new date using the numeric keys on the keypad The cursor is positioned under the first editable field and advances automatically gt To change the date format press the soft key beneath MMDDYY or DDMMYY The display automatically updates to reflect the new format Press ENTER to return to the EDIT SETUP INFORMATION menu 3 Tochange the current time and or the time format select TIME from the EDIT SETUP INFORMATION menu The TIME entry screen will appear 47 PM 12HOUR 24 HOUR AM PM gt Enter the time using the numeric keys on the keypad The cursor is positioned under the first editable field and advances automatically gt Tochange the time format select 12HOUR or 24HOUR then AM or PM The display automatically updates to reflect the new format gt Press ENTER to return to the SELECT UTILITY OPTION menu BioTek Instruments Inc Utility 93 Editing the Wavelength Table Y The wavelength table contains six user defined wavelength values that are presented for selection when defining an assay See Meas Selection on page 45 To edit the wavelength table 1 From the Main Menu select UTIL gt SETUP At the EDIT SETUP INFORMATION menu select LAMBDA The EDIT WAVELENGTH screen will appear EN TER LAMBDAHd 1 WAVELENGTH gt Enter the correct wavelength for each of the six positions using the numeric keys The range is from 200 to 999 nm gt C
59. 07 Set up various global configuration options such as date and time report output and plate reading preferences page 86 READY 01 22PM 01 23 04 READ DEFINE REPORT UTIL SELECT UTILITY OPTION TESTS SETUP OUTPUT READ PRINT REPORT RESULT MAP ASSAY LIST SELECT ASSAY NUMBER 03 NAME BIO CELL READ SELECT ASSAY NUMBER 03 NAME BIO CELL READ Figure 14 Options available from the Main Menu UQuant Operator s Manual 40 Chapter 4 Operation Define The Main Menu option DEFINE allows you to define the data acquisition and reduction parameters for a new assay or modify preprogrammed assays stored in memory Table 1 Table 1 Preprogrammed Assay List Spectral Scan assay EDTA test assay 22 01 Raw_OD assay A320 Scattering assay HRP TMB assay 02 Raw_OD w blk assay Lowry Protein assay Alk Phosphatase assay 03 Bio Cell Read Bradford Protein assay 22 HRP TMB S assay RNA Quantassay 18 HRP5ASassay 29 WST Lasay Oligo Quantassay I HRP ABTS assay 30 Methylene Blue assay E N cic 10 1 Startat the Main Menu and select DEFINE to display the SELECT ASSAY NUMBER screen SELECT AS SAY NUMBER NAME H B S AG 1 2 Select an assay to define or modify then press ENTER See Selecting an Assay to Define on the next page for detailed instructions The EDIT ASSAY NAME screen will appear If you are modifying selecting a PANEL assay 99 see page 78
60. 2 14 bul Es Solution ml vmm d 3 Create a percentage dilution series beginning with 10076 of the concentrated stock solution in tube 1 90 of the concentrated solution in tube 2 80 in tube 3 and so on to 10 in tube 10 4 Dilute using amounts of the remaining 0 0576 solution of deionized water and Tween 20 as shown in Table 5 UQuant Operator s Manual 124 Chapter 5 Instrument Qualification Plate Preparation 5 Pipette 200 ul of the concentrated solution from tube 1 into each well of the first column A1 to H1 of the microplate 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 5 on the previous page as a model for calculating the expected absorbances of a series of dilutions given a different absorbance of the original solution After pipetting the diluted test solution into the microplate and before reading the plate we strongly recommend shaking the plate for four minutes This will allow any air bubbles in the solution to settle and the meniscus to stabilize If a plate shaker is not available wait 20 minutes after pipetting the diluted test solution before reading the plate Read Plate Read the microplate prepared above five times using Normal Read Mode dual wavelength at 450 nm with 630 nm as the blank
61. 99 Pax 49 0 TLS6 968 11 1 All Others Tel 802 655 4040 Fax 802 655 3399 email TAC biotek com Product support center www biotek com service Figure 16 Sample output for the System Test Autocal Analysis sheet 2 of 2 If the reader fails the System Test it will not perform a read until the error condition is cleared See Chapter 7 Troubleshooting and Error Codes for a list of possible error codes that may be displayed if the System Test indicates a failure Photodiodes The Optics portion of the System Test confirms that the reading channel has adequate signal range without saturating the electronics Light Bulb The Optics test also indicates if the light bulb is within operational limits The System Test includes subtests for Gain Values and Resets Air Dark Value Noise Test and Autocal Analysis The data and results for these tests can be reviewed on the System Test report BioTek Instruments Inc Qualification Procedures 105 Note The lamp life is rated at an average of 1 billion flashes The lamp should last beyond the useful life of the uQuant reader If there is a problem with the lamp the intensity may drop and the runtime self check would detect a low signal level that would appear on the instrument s display Contact BioTek s Technical Assistance Center for assistance if the lamp needs to be replaced Refer to Chapter 1 for contact information Gain Values and Resets The energy from t
62. Absorbance Test Plate UQuant Operator s Manual 100 Chapter 5 Instrument Qualification Qualification Procedures You may use the tests outlined in this section to confirm initial and ongoing performance of the uQuant Set up the Quant according to the instructions in Chapter 3 Installation Confirm that the instrument powers up and communicates with peripherals for the Installation Qualification After Installation Qualification conduct the Operational Qualification Tests lt Note An Installation Operational Performance Qualification IQ OQ PQ package PN 7270514 for the Quant is available for purchase Contact your local dealer for more information Your Quant reader was fully tested at BioTek before shipment and should operate properly upon initial setup If you suspect that problems may have occurred during shipment if you reshipped the instrument or if regulatory requirements dictate that Performance Qualification Testing is necessary you should perform the following tests After the initial confirmation of operation you should perform the Absorbance Plate Test monthly and Liquid Testing semiannually e The System Test described on the next page verifies proper gains bulb operation and low electronic noise e The Checksum Test page 107 compares the onboard software with internally recorded checksum values to ensure that no corruption has occurred e The Absorbance Plate Test page 108 confirms the optical accu
63. CDC assumes any liability for the adequacy of these solutions and methods Each laboratory must ensure that decontamination procedures are adequate for the Biohazard s they handle Wear prophylactic gloves when handling contaminated instruments Gloved hands should be considered NA contaminated at all times keep gloved hands away from eyes mouth nose and ears Eating and drinking while decontaminating instruments is not advised Mucous membranes are considered prime entry routes for infectious agents Wear eye protection and a surgical mask when there is a possibility of aerosol contamination Intact NA skin is generally considered an effective barrier against infectious organisms however small abrasions and cuts may not always be visible Wear protective gloves when performing the decontamination procedure BioTek Instruments Inc Decontamination Procedure 135 Tools and Supplies e Sodium hypochlorite NaCIO or bleach e 70 isopropyl alcohol as an alternative to bleach e Deionized or distilled water e Safety glasses e Surgical mask e Protective gloves e Lab coat e Biohazard trash bags e 125 ml beakers e Clean cotton cloths Procedure The bleach solution is caustic wear gloves and eye protection when handling the solution Do not immerse the instrument spray it with liquid or use a wet cloth Do not allow the cleaning solution to run into the interior of the instrument If this happens co
64. Microplate Spectrophotometer UQuant Operator s Manual ssBiolek wQuant Microplate Spectrophotometer Operator s Manual May 2006 2006 Part Number 7271000 Revision I BioTek Instruments Inc Notices Preface BioTek Instruments Inc P O Box 998 Highland Park Winooski Vermont 05404 0998 USA All Rights Reserved 2006 BioTek Instruments Incorporated No part of this publication may be reproduced transcribed or transmitted in any form or by any means electronic or mechanical including photocopying and recording for any purpose other than the purchaser s use without written permission of BioTek Instruments Inc Trademarks BioTek is a registered trademark and wQuant Extensions Gen5 KC4 and KCjunior are trademarks of BioTek Instruments Inc Windows is a registered trademark of the Microsoft Corporation All other trademarks are the property of their respective holders Restrictions and Liabilities Information in this document is subject to change and does not represent a commitment by BioTek Instruments Inc Changes made to the information in this document will be incorporated in new editions of the publication No responsibility is assumed by BioTek for the use or reliability of software or equipment that is not supplied by BioTek or its affiliated dealers BioTek Instruments Inc Contents iii Contents Niels ec rara i CONTONES e
65. NCTION The following functions can be used in formulas ALOG10 Anti Log Base 10 LOG Log Absolute Value PWR SORT EXAMPLES LOG10 Log Base 10 ALOG Anti Log ALOG 0 69314718 2 Log10 2 0 301029995 ALOGIO Anti Log Base 10 LOG Log LOG 2 0 69314718 ALOG10 0 30102995 2 AB Absolute Value AB 1 1 SQRT Square Root SQRT 2 1 4142 PWR Power 10 PWR 2 100 uQuant Operator s Manual 66 Chapter 4 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 63 for instructions on selecting an assay and accessing the SELECT VALIDATION TYPE screen SELECT VALIDATION TYPE CONTROL ASSAY B LANK 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 it applies 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 One blank validation formula can be defined Up to four control validation formulas can be defined Define the plate map via DEFINE gt
66. RE P 1 CO 2 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 uQuant Operator s Manual 62 Chapter 4 Operation Defining FORMULA The uQuant supports three types of formulas Cutoff Transformation and Validation 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 1 Start at the Main Menu and select DEFINE 2 Select the assay then press ENTER The DEFINE options screen will appear DEFINE ME TH OD M A P FORMULA CURVE 3 Select FORMULA The SELECT FORMULA TYPE screen will appear Calculation Structure During data reduction formulas are processed in the order shown below The number of permitted formulas of each type is shown as well e Blank Validation 0 1 e Control Validation 0 4 e Assay Validation 0 4 e Transformations 0 1 e Cutoff Formulas 0 1 D To capture and manipulate the raw data using 384 well microplates with the uQuant you must use Gen5 M KC4 or KCjunior M Gen5 KC4 and KCjunior are PC based software programs you can use to set up your assay communicate with the uQuant to run the assay and then manipulate the raw data that is automatically retrieved from the reader Refer to Appendix B for additional information BioTek Instruments
67. RE P 1 GO 1 e The sample well locations originally defined in the assay will be presented If desired use the keypad to enter new well locations for each sample replicate e Press ENTER again to advance to the next replicate BioTek Instruments Inc Reading a Microplate 87 Beginning the Plate Read When the following screen appears on the display the reader is ready to read a plate PLACE PLA TIE I N CARRIER AND PRESS lt RE AOD gt K E Y e Before reading the plate ensure that the printer is connected turned on and full of paper e Place the plate in the carrier and press the READ key to initiate the plate read e After the read is complete data reduction will be performed Calculating Results and then the reports will print Generating Reports e To halt the read in progress press the STOP key UQuant Operator s Manual 88 Chapter 4 Operation Printing Reports Reports are automatically generated after a plate has been read see Output Options in Chapter 3 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 Y Note See Appendix C for sample reports 01 22 PM 01 23 04 DEF INE REPORT U TI L RINT REPORT E SULT MAP ASSAY LIST e Select RESULT to print an exact copy of results from the plate reading the 8
68. RESULTS O e Ensure that the printer is connected turned on and filled with paper e Press YES to print reports or NO to return to the Main Menu 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 SELECT AS SAY NUMBER NAME HBS AG 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 SELECT AS SAY NUMBER NAME HBS AG 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 and then select LIST to print the report UQuant Operator s Manual 92 Chapter 4 Operation Utility The Utility option allows you to set up the date and time specify your data output and report options and select your read options Setting the Date and Time To set the current Date and Time and or change their formats 1 From the Main Menu select UTIL gt SETUP The EDIT SETUP INFORMATION menu will appear SETUP INFORMATION T IME LAMBDA MORE 2 Select DATE The DATE entry screen will appear
69. Report Sheet 1 of 2 195 UQuant Operator s Manual 196 Appendix C Report Format Assay List Version v2 00 1 iO O J2 OY Oi 4i C P9 FB OO Figure 41 Assay Detail Report Sheet 2 of 2 SPECTRAL SCAN Raw OD Raw OD w blk Bio Cell Read KFactor Bio Cell DNA Quant 260 280 Ratio RNA Quant Oligo Quant Protein Quant Phenol EDTA A320 Scattering Lowry Protein Bradford Protein BCA Protein B galactosidase NADH HRP 5 Test AS HRP ABTS HRP OPD S HRP OPD HRP TMB S HRP TMB Alk phosphatase B galact ONPG Urease MTT ATT WST 1 Methylene Blue Open Open Open Open Open Open Open Open Open Open Open Open Open Open Open Open Open Open Open Open Open Open Open Open Rpen Open Open Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay Assay 01 02 03 04 05 06 07 08 09 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 BioTek Instruments Inc 197 Operator ID Notes 11 21AM 06 29 98 SPECTRAL SCAN 400nm 600nm 2nm AT WELL LOCATION HOG Min OD 0 713 530nm Max 0D 1 926 402nm 1 926 9 i ttt 1 683 Salah aaa aat aa Lee DEERE SETTE TETEE TETEE TEREE Ooo Cte ALL c oo p or ror Orne 400 410 420 430 440 450 460 470 480 490 500 510 520 530 540 550 560 570
70. See Delta Air Dark on self test This error indicates one of the following situations e During system test the Delta was lt 8000 for one of the wavelengths e During a spectral scan or blank read the flash on value minus flash off lt 8000 normal mode lt 8000 calibrated resets 8 for sweep mode Probable Causes e Lamp failure alignment or lamp power supply e Absorbance channel analog PCB The order sorting filter wheel is jammed and not aligning the correct filter to line up with the light path the filter is degraded and not passing enough light energy or the filter is blocking the light Damaged optic spray e Assay code corrupted or not downloaded Failed configuration checksum test This error indicates that during self test or at the end of a plate read the checksum calculated for configuration flash memory page 0 doesn t match saved checksum Probable Cause e The flash memory on the 7270400 PCB is defective or corrupt The basecode software may need to be redownloaded Failed configuration checksum test This error indicates that during self test or at the end of a plate read the checksum calculated for configuration flash memory page 1 doesn t match saved checksum Probable Causes e The flash memory on the 7270400 PCB is defective or corrupt The basecode software may need to be redownloaded Configuration data missing Indicates the configuration data was missing at the beginning
71. Sia adidas Package Contents iia Optional ACCESS ONES arcada Eds SPECINCALIONS ssaniieeasrectasiaennnnccraneeaninarranuantaceana iia Product SUP DOL A SEVICE alias 2 Chapter 1 Introduction Introducing the pQuant The Quant is a single channel microplate spectrophotometer for research and development and in vitro diagnostic use designed to automatically perform endpoint analysis In the European Union This product may only be used for Research and Development and other nonclinical purposes In all other jurisdictions This product may be used for Research and Development and in vitro Diagnostic purposes The reader can measure the optical density of solutions in 6 12 24 48 96 or 384 well microplates The data files created can be printed in report form or stored as data files for a variety of ELISA based applications The reader features superior optical specifications with an extended dynamic range of up to 4 000 absorbance units in some read modes The wavelength range is from 200 nm to 999 nm Kinetic analysis can be performed using computer control Assay definitions consisting of protocols templates and formulas and the data they produce are managed by an onboard processor via a 2 line x 24 character LCD screen and keypad The uQuant is designed to serve as a stand alone system or as part of a larger laboratory data network sending receiving and manipulating assay data as needed Data can be stored onboard pri
72. ZONE 0 20 0 200 CUTOFF 1 1 000 Hn LE SMP SMP lt CUTOFF CUTOFF GREYZONE AND SMP gt CUTOFF CUTOFF GREYZONE EQUIV SMP SMP gt CUTOFF CUTOFF GREYZONE NEG SMP SMP lt CUTOFF CUTOFF GREYZONE POS PC PC gt 01 Well 1D 81kd00 CalcOD Call Pred Conc RSLT Std Dev CV Notes BU PC 0 119 0 119 VAL AVE 0 114 0 114 0 007 6 163 cor SNPO00 0 20 0 200 POS DOI SNP0002 030 030 POS E01 SNP0003 2 205 2 205 NEG F01 SNPO004 28 2 811 NEG G01 SNP0005 3 499 3499 NEG HOt SMPODOS 125 3 225 NEG A02 SMP0007 0 085 0 085 POS B02 SHPO0O8 1 054 1 054 EWN C02 SNPO009 1 092 1 092 EQUIV D02 SMPOO1 2 075 205 NEG E02 SNP0011 2 064 2 004 NEG FO SNP0012 124 3 21 NEG 602 SNPO013 3 663 3 00 MEG HO2 SHPOO14 1 045 1 045 EQUIV A03 SHPOO15 0 00 0 080 POS Figure 37 Samples with calls on Column Report BioTek Instruments Inc 193 Bio Tek Instruments Assay Open Assay 04 Date 03 16 04 Lot Time 01 05 34PM Operator Wavelength 405 Temp Plate ID COMMENTS Interpretation of Results GREYZONE 0 20 0 200 CUTOFF 1 1 000 SMP SMP lt CUTOFF CUTOFFFGREVZONE AND SMP gt CUTOFF CUTOFF GREYZONE EQUIV SMP SMP gt CUTOFF CUTOFFFGREYZONE NEG SMP SMP lt CUTOFF CUTOFFSGREYZONE POS PC PES uH Well ID Cal Pred Conc RSLT Std Dev A01 RC i 0 109 801 PC i 0 119 AVE 0 114 0 007 6 163 End of Report Fi
73. a tablets In a beaker mix 50 ml of the PBS solution prepared from the Sigma tablets with 10 mg of the B NADH powder and mix thoroughly This is the Sigma PBS Solution BioTek Instruments Inc Procedure Liquid Testing 129 1 Check the absorbance of a sample of buffer solution A or B at 340 nm on the microplate reader This solution which will be referred to as the 100 Test 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 If low adjust up by adding D NADH powder until the solution is at least at the lower end of this range Do not adjust if slightly high 2 Carefully prepare a 75 Test Solution by diluting 15 ml of the 100 Test Solution gt gt If using the Sigma PBS Solution use 5 ml as the diluent If using the 10x Concentrate PBS Solution mix one part of the concentrate with nine parts of DI water Use 5 ml of this solution as the diluent 3 Carefully prepare a 50 Test Solution by diluting 10 ml of the 100 Test Solution gt gt If using the Sigma PBS Solution use 10 ml as the diluent If using the 10x Concentrate PBS Solution mix one part of the concentrate with nine parts of DI water Use 10 ml of this solution as the diluent 4 Pipette the three solutions into a new 96 well microplate gt 150 ul of the 100 Test Solution into all wells of columns 1 and 2 150 ul o
74. ad Mode 0 000 to 2 000 OD 1 0 010 OD Rapid Mode or Spectral Scanning 2 000 to 2 5000 OD 3 0 010 OD Rapid Mode or Spectral Scanning Measured with neutral density filters calibration plate O 405 450 490 550 630 690 nm 0 000 to 2 000 OD 1 Normal Read Mode 2 000 to 3 000 OD 3 Normal Read Mode 0 000 to 2 000 OD 1 Rapid Mode or Spectral Scanning 2 000 to 2 500 OD 3 Rapid Mode or Spectral Scanning Measured 405 nm using yellow food dye 0 000 to 2 000 OD 1 0 005 OD Normal Read Mode 2 000 to 3 000 OD 3 0 005 OD Normal Read Mode 0 000 to 2 000 OD 1 0 005 OD Rapid Mode or Spectral Scanning 2 000 to 2 500 OD 3 0 005 OD Rapid Mode or Spectral scanning Measured 405 nm using yellow food dye Hardware and Environmental Specifications Display Light Source Dimensions Weight Environment Humidity Power Supply 2 line x 24 character LCD display Xenon flash 42 cm x 38 cm x 18 cm 16 deep x 15 wide x 7 high 11 34 kg 25 lb maximum Operational temperature 18 40 C 64 4 104 F 10 to 85 noncondensing External 24 V power supply compatible with 100 240 V 10 O 50 60 Hz BioTek Instruments Inc Product Support amp Service 7 Product Support amp Service A superior support staff backs all of BioTek s products If your instrument s or software ever fails to function perfectly if you have questions about how to use or maintain them or if
75. al components Caution Environmental Conditions Do not expose the system to temperature extremes For proper operation ambient temperatures should remain between 18 40 C Performance may be adversely affected if temperatures fluctuate above or below this range Storage temperature limits are broader Caution Sodium Hypochlorite Do not expose any part of the instrument to the recommended diluted sodium hypochlorite solution bleach for more than 20 minutes Prolonged contact may damage the instrument surfaces Be certain to rinse and thoroughly wipe all surfaces Caution Power Supply Only use the power supply shipped with the instrument Operate this power supply within the range of line voltages listed on it Caution Shipping Hardware A carrier shipping block must be removed before operating the reader The shipping block must be reinstalled before repackaging the reader for shipment See Chapter 3 Installation Caution Warranty Failure to follow preventive maintenance protocols may void the warranty See Chapter 6 Maintenance and Decontamination Caution Disposal This instrument contains printed circuit boards and wiring with lead solder Dispose of the instrument according to Directive 2002 96 EC on waste electrical and electronic equipment WEEE BioTek Instruments Inc Hazards and Precautions XIX C Based on the testing described below and information contained herein this instrument bears
76. ally coded Failure Mode Effects Analysis FMEA criteria established by BioTek A system test should then be initiated to try to retrieve an error code from the reader Glossary of Terms e Air Blank A full light reading through a filter with no plate in the light path e Dark Current A reading taken with the light blocked to measure background light levels in the reading chamber Also used as a measure of background electronic noise within the measurement circuit e Gain An automatic electronic adjustment to the measurement circuit The gain adjustment compensates for changing light levels or filter variations For example if the lamp output decreases slightly the gain will increase to make up the difference e Axis Refers to a motor for the filter wheel or plate carrier e Offset A numerical limit usually a range For example if the gain fails an offset test it may be too high or too low BioTek Instruments Inc Error Codes 139 Error Codes An error code is displayed on the instrument as a four digit identifier The first character will be 0 1 2 or A Note If an error code is displayed run a System Self Test for diagnostic purposes e 0 1 or 2 indicates a noncritical error and the instrument will respond to keypad input See General Errors on the next few pages for more information Motor or Optical Sensor Channel 2 Monochromator Order Sorting Filter Wheel DS e A indicates a mor
77. and set them aside Power Supply Poly Bag Bottom End Ca L bo Printer Cable LE x em TT UV Sample A Power Cord Plate Pack Dust Cover Figure 3 Removing the reader and accessories from the outer shipping box BioTek Instruments Inc Unpacking and Inspecting the uQuant 17 4 Place the instrument on a stable surface 5 Use the slotted screwdriver to remove the two black dress screws from the rear panel of the instrument Figure 4 Set aside these screws Black Dress Screws Corn 4 Figure 4 Removing the black dress screws from the rear panel UQuant Operator s Manual 18 Chapter 3 Installation 6 Turn the instrument around and open the front door While holding the carrier in place use a Phillips head screwdriver to remove the shipping block and mounting hardware Figure 5 and set them aside IMPORTANT Do NOT remove the two black dress screws that secure the carrier sliding block Figure 5 to the carrier 7 Insert the two black dress screws that were removed from the rear panel into the shipping block mounting holes Front Door Remove Shipping Block and Mounting Hardware replace with black dress screws from rear panel Black Dress Screws Do NOT Remove Carrier Sliding Block Figure 5 Removing the shipping block and inserting the black dress screws from the rear panel BioTek Instruments Inc Unpack
78. apter 4 Operation MAP screens appear in the order that they were defined in the assay If the assay has a closed variable the screen for this variable is omitted DEFINE METHOD MA P FORMULA CURVE e At the DEFINE options screen select MAP to begin the plate map process Map Generation Map Generation represents the method by which blanks controls standards and or samples are assigned to specific locations on the plate M A P GENERATION MANUAL AU TO MANUAL e Select AUTOMATIC PLATE MAP GENERATION to instruct the software to automatically generate a plate map after the blanks controls standards and or samples have been defined e Select MANUAL PLATE MAP GENERATION to indicate that the well assignments will be performed manually by the user at Define and or Read time e Press ENTER to save the selection and continue Use the SHIFT CLEAR keys to clear any previously defined map BioTek Instruments Inc Define Method Map Formula and Curve 49 Mapping Direction This option allows you to specify how the blank control standard or sample groups will be mapped on the plate The well types can be listed in column format down or in row format across The currently selected Mapping Direction appears on the top line of the display and the available options appear on the bottom MAPPING DIRECTION DOWN DOWN ACROSS Select DOWN to map down the column Select ACROSS to map across the r
79. ard concentration values The range is 00001 to 999999 The entry cannot exceed six 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 the next standard UQuant Operator s Manual 58 Chapter 4 Operation Valid Well Locations When defining the replicate locations only the alpha keys are active for the first character and numeric for the second and third characters Valid characters and numeric entries are based on the selected plate geometry The following table lists acceptable entries for well locations based on plate geometry Plate Type Wl 12 Well 24 Well 48 Well 96 Well 384 Well ZX Number of Controls The NUMBER OF CONTROLS screen allows you to enter the number of control groups that will be used in the assay Any previously defined value will appear on the display e Use the numeric keys to enter the NUMBER OF CONTROLS 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 l
80. as 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 Transformation Variable TVAR For more complex transformations a Transformation Variable IVAR 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 just to the sample wells SMP SCOPE VARIABLE D BioTek Instruments Inc Define Method Map Formula and Curve 69 If SMP is chosen The transformation formula will be applied to the sample wells only SMP and any other well identifiers BLK PC NC STD etc defined will become available as MAP options when building the transformation formula 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 If OD is chosen The formula definition screen will appear so that you can define a formula for use within the transformation formula 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 transformati
81. aseseebidiesaotssattces 20 Electrical Gonnectlors ssi estas bet bp ia 20 Power Up and System Test eseeesseeene nnnm nnne nnne nnn 21 jouant Main MERU soutien RA DS Cua T E e 21 Adjusting the Wavelength Table eee ne mmn 22 Connecting a Printer to the UQUANL cece cece eeee eee eeeeeeeeeeeaeees 24 Optional Setting Up the Serial Port for Communications 26 Attaching the Cable 26 Installing the PC Software 20000 26 Setting Communication Parameters cece cece cece ee ee eee eeeeeeeeees 26 Repackaging and Shipping the puQuant een n 28 14 Chapter 3 Installation Unpacking and Inspecting the uQuant D Important Save all packaging materials If the uQuant is shipped to the factory for repair or replacement it must be carefully repackaged according to the instructions on pages 28 through 33 using the original packing materials PN 7273000 The individual packaging materials contained in PN 7273000 are listed in the table below Using other forms of commercially available packing materials or failure to follow the repackaging instructions may void your warranty If the original packing materials have been damaged replacements are available from BioTek The wQuant and its accessories are securely packaged inside custom designed shipping materials This packaging should protect the instrument from damage during shipping Inspect the shipping box packaging instrument
82. assay is selected one or more informational prompts may be presented depending on preferences selected in UTIL gt READ whether or not the assay specifies manual mapping or if the assay was created or downloaded from BioTek s Extensions Define Reader Protocol software e Prompts enable via UTIL gt READ can include ENTER NUMBER OF SAMPLES PLATE ID and ENTER SAMPLE ID e If the assay specifies manual mapping prompts for information will include the locations for the sample wells e If running a custom assay such as one that was created using Extensions software typical prompts might include gt gt Vv Vv Y Y VW vv The number of samples Standard concentrations Assay ID Fill pattern Blank method First well location Replicate count for each well type Wavelength mode Report preferences etc BioTek Instruments Inc Reading a Microplate 85 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 predefined in the assay but if this is a custom assay you may also be prompted to enter the replicate count EN TER NUMBER OF SAMPLES 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 tha
83. ation for Liquid Tests Nos 1 and 2 119 Table 5 Test TUbe DIIULIOL Siero sspe vea bti Be vir eost PER PREX Lea rUE POTATO ES PVP 123 Table 6 PBS 10X Concentrate Solution eene 128 Table 7 Serial Cable Pinout Description esee mnn nn 175 UQuant Operator s Manual X Preface Contacting BioTek Instruments Inc BioTek Instruments Inc P O Box 998 Highland Park 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 customercare 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 tac biotek com European Coordination Center BioTek Instruments GmbH Kocherwaldstrasse 34 D 74177 Bad Friedrichshall Germany Internet www biotek de Phone 49 0 7136 9680 Fax 49 0 7136 968 111 E Mail info biotek de BioTek Instruments Inc Document Conventions XI Document Conventions This manual uses the following typographic conventions Example Description A A Warning A Warning indicates the potential for bodily harm and tells you how to avoid the problem These icons call attention to important safety notes Caution A Caution indicates potential damage to the instrument and tells you how to avoid the problem DEFINE Text in COURIER font represe
84. aulty Analog PCB or faulty internal grounding This type of problem would require factory servicing Autocal Analysis The Autocal Analysis is run only at the factory to calibrate the carrier axis of the instrument The results of this test are included in the System Test report see page 104 In the field the alignment test of the Absorbance Plate Tests is used to verify that the carrier positioning is correct BioTek Instruments Inc Qualification Procedures 107 Checksum Test This test also runs automatically when the reader is turned on The test compares the software to the internally recorded checksum values to ensure that the programming has not been corrupted It verifies the checksum part numbers and versions of software currently loaded onto your reader If there are any errors during the power on checksum test they will be displayed 1 Select CHKSUM at the SELECT TEST screen to run the Checksum Test The information displayed will resemble the following 72 70201 VERS ION COD E CHECKS UM gt The initial checksum test display will show the onboard base code software part number version number and checksum After a few moments a second screen will display 7270203 FW gt The second checksum test display will show the assay configuration software part number and version number After a few moments the Main Menu will reappear UQuant Operator s Manual 108 Chapter 5 Instrument Qual
85. blems gt If the test is not successful and you receive an error message refer to the Problems section below or to the Troubleshooting section of Gen5 s Help system for assistance gt Gend Secure only An Audit Trail dialog will appear after exiting Reader Configuration whenever you add modify or delete a reader Enter any comments if desired then click Close If Gen5 fails to communicate with the reader try the following troubleshooting suggestions Confirm that the correct Reader Type was selected in step 9 Try a different COM port Check the serial cable connections Ensure that the cable is properly attached to the port defined in step 10 and is not a Null cable If this is suspected add another Null and try again Confirm that the reader has passed its Self Test The reader will not communicate if it fails an internal system test If the test still fails refer to the Troubleshooting section in Gen5 s Help system for further assistance Getting Started with Gen5 The following instructions briefly show you how to define and perform a Quick Read in Gen5 File New Experiment Default Protocol It s called Quick because you can perform a reading without having to take the time to create a new protocol If the reading is part of an experiment or assay that you will perform numerous times you will need to create a new protocol File New Protocol Refer to Gen5 s Help system early and often to learn how
86. ce Plate Test parameters report as FAIL confirm that the standard values on the test plate Standards Certificate match the values on the printout If not correct and retest If the test still fails contact BioTek s Technical Assistance Center refer to Chapter 1 for contact information Please have a copy of the test and the reader s serial number available when you call e Mechanical Alignment The Test Plate has several groups of precisely machined holes to confirm the mechanical alignment of different microplate readers The amount of light that shines through these holes indicates whether the reader is properly aligned A reading of more than 0 015 OD for any of the designated alignment wells indicates that the light is being clipped and the Quant may be out of alignment If the reader fails the alignment test review the following possible problems and solutions gt Ensure that the Test Plate is correctly seated in the microplate carrier Check the four alignment holes to ensure that they are clear of debris gt Check the microplate carrier to ensure that it is clear of debris If these remedies are ineffective call your BioTek representative or BioTek Technical Support e Accuracy and Linearity The Test Plate contains neutral density glass filters that have assigned OD values at several wavelengths Since there are several filters with differing OD values the accuracy across a range of ODs can be established Once it i
87. ce data corrected reference dark offset The ratio will fail if lt 0 5 or gt 2 0 e During a spectral scan or blank read the flash on value minus flash off lt 500 normal mode or lt 500 calibrated resets 8 for sweep mode Probable Causes e Lamp failure alignment or lamp power supply Reference channel analog PCB or absorbance channel analog PCB or the cable in between e The order sorting filter wheel is jammed and not aligning the correct filter to line up with the light path the filter is degraded and not passing enough light energy or the filter is blocking the light e Damaged optic spray Measurement channel correction outside limits See criteria below This error indicates that during a spectral scan the flash on value minus flash off lt 8000 normal mode lt 8000 calibrated resets 8 for sweep mode Probable Causes Lamp failure alignment or lamp power supply Reference channel analog PCB or absorbance channel analog PCB or the cable in between The order sorting filter wheel is jammed and not aligning the correct filter to line up with the light path the filter is degraded and not passing enough light energy or the filter is blocking the light Damaged optic spray See the next page for more situations UQuant Operator s Manual 160 Chapter 7 Troubleshooting and Error Codes General Errors Continued Measurement channel correction outside limits See criteria below
88. channel failed offset range See noise Max lt 20000 and noise Min gt 10 on the system test During self test the background electronic signal detected is outside of acceptable limits at maximum gain when blocking the light Probable Causes e If noise Max is gt 20000 gt The photodetector is too noisy and defective Analog PCB is defective A faulty analog PCB or faulty internal grounding may cause internal electronic noise There may be an ambient light leak Ensure that the plate carrier door is properly closed Electrical noise may be penetrating the measurement chamber The bottom and top shrouds are part of the electrical shielding Verify that the shrouds are installed and properly fastened e If noise Min is lt 10 gt The photodetector is not connected or the optic spray is damaged or defective producing a noise reading of zero Analog PCB is defective BioTek Instruments Inc Error Codes 153 General Errors Continued 0900 The absorbance reference channel Dark current value failed See Dark on the system test See criteria below This error indicates one of the following situations e The reference channel failed during a read or spectral scan for one of the following reasons Dark value was lt 100 during a spectral scan using 8 flashes and 8 resets during sweep mode Dark value was lt 100 during a spectral scan using 8 flashes and the number of rese
89. ct a Universal Plate ID Wavelength enter the Reader Serial Number then click Run Test For wavelength accuracy While performing the steps above click Perform peak wavelength test in Gen5 or KCjunior or click Peak absorbance search in KC4 BioTek Instruments Inc Calibration Plate Analysis 12 30PM 01710706 Operator ID Notes Alignment Results B2 0 000 PASS Accuracy Results C01 0 147 Dn La PASS D04 Lx dd Ld PASS STANDARD DATA RESULT Repeatability Results COT Q3 0 141 PASS D04 A led Zu PASS READ1 READ2 RESULT Spectral Scan Results 580nm 1 933 586nm 2 581nm 1 956 587nm 2 582nm 2 059 588nm 2 583nm 2 253 589nm 2 584nm 2 511 590nm 2 585nm 2 785 Test Plate Standard 587nm PASS Calculated Peak 586nm Read Mode B12 0 000 PASS E02 0 618 O56 13 PASS E02 0 613 Du 6116 PASS VO cu s 916 2334 186 Normal Gl 0 000 PASS F05 PEN M ZOO PASS FOS ZOO PEL PASS Lambda G03 ES 1 129 PASS G03 Dg EZ8 ANAL D PASS 40 Gl H06 AO 1 69 PAS H06 109 1359 PAS Qualification Procedures 113 9 1 0 000 PASS il 4 S 4 4 S Figure 18 Sample Absorbance Plate Test Report Note The format varies depending on the software used to run the test Quant Operator s Manual 114 Chapter 5 Instrument Qualification Absorbance Plate Test Results and Troubleshooting Tips If any of the Absorban
90. ction Wavelengths Biazell E Options First well jan Last well P24 Monitor Wells Option m e Figure 30 Reading dialog 10 When you have finished defining the parameters click OK KC4 will automatically validate the defined parameters If any parameters are incorrect or incomplete follow the instructions provided by KC4 for correcting the parameters 11 Select Data Read Plate The Plate Reading dialog will appear 12 Enter any desired information place the 384 well plate on the carrier then click START READING to begin the plate read gt The plate will be read and then the raw data results will be displayed in KC4 gt To analyze manipulate or print results Protocol parameters should be defined Refer to KC4 s Help system or User s Guide for instructions BioTek Instruments Inc Reading a 384 Well Plate in KCjunior 185 Reading a 384 Well Plate in KCjunior 1 Start KCjunior 2 At KCjunior s main screen choose Setup Reader 1 or Reader 2 Results Protocols Utilities Setup Current Reader View Help Reader 1 Reader 2 Configuration Figure 31 Setup Reader dialog 3 The Reader Setup dialog will appear Select the Quant reader for the Reader Type Reader Setup 1 E X sienne Carcel Help Get Comm Defaults Test Communications GetBasecode Version Communications Settings Com Port
91. ctral scan meaning the wavelength has not been calibrated This includes the gain test data looking for the resets to be lt 1 or gt 8 to fail Probable Causes Memory corrupt The calibrate before read check box is not selected and this wavelength has not been used prior to this error The gain test was skipped or failed for this wavelength Wavelength calibration data missing during an absorbance read See criteria below Wavelength data is missing prior to an absorbance read meaning the wavelength has not been calibrated This includes self test and gain test looking for the resets to be lt 1 or gt 8 to fail The last number is the lambda table position number Probable Causes e The wavelength was not calibrated prior to the read e The th UQuant Operator s Manual 158 Chapter 7 Troubleshooting and Error Codes General Errors Continued Wavelength not found in table The specified wavelength is not detected in the instrument s filter table The last number is the filter set number in the assay protocol Probable Causes Wavelength not entered correctly or missing in filter table Wavelength entered correctly in PC software but never sent to reader Verify the lambda table has the wavelengths loaded into the instrument from the controlling PC software Compare the contents of the lambda table with the software s filter table Reference channel correction outside limits See criteria below
92. d uQuant Operator s Manual 24 Chapter 3 Installation Connecting a Printer to the Quant D Connect the printer to the wQuant only if you are running the instrument in standalone mode If you are using Gen5 KC4 or KCjunior software skip this step and go to page 26 Setting Up the Serial Port for Communications The uQuant has a parallel port LPT1 to allow connection to Epson compatible printers or HP Deskjet printers See page 93 under Specifying Data Output and Reporting Options for more information The port is illustrated in Figure 7 The parallel port requires a 25 pin D sub connector A parallel cable PN 71072 designed to connect the reader to a printer is provided with the u Quant To attach a printer to the Quant 1 2 3 4 5 6 D Turn the reader off Place the printer in a location adjacent to the unQuant Attach one end of the cable to the parallel port on the printer Attach the other end of the cable to the parallel port on the uQuant Tighten the securing screws on both ends of the cable Turn on the reader and then turn on the printer To avoid system instability be sure to connect the printer to the reader before powering up the reader BioTek Instruments Inc Setting Up the uQuant 25 Serial Port Parallel Port Printer Port Power Supply Jack Figure 7 Connectors for printer parallel computer serial and power supply UQuant Operator s Manua
93. different location than it had previously This error indicates that filter three has saturated the reference channel UQuant Operator s Manual 146 Chapter 7 Troubleshooting and Error Codes General Errors Continued 0503 Probable Causes Cont monochromator Not Found Home Flash bulb skipped a flash due to poor lamp connection or defective power supply Order sorting filter wheel is jammed and is not aligning the through hole to the light path Monochromator is defective Analog PCB intermittently failed Reference Channel The monochromator mirror grating is damaged Analog PCB intermittently failed Missed flashes or erratic flash lamp The order sorting filter wheel is jammed and not aligning the correct filter to line up with the light path or the through hole is allowing white light to pass Voltage to the lamp has increased because the lamp power supply has failed or the motor power supply PCB sent the incorrect voltage request to the lamp power supply Light beam saturated too much light Air measurement channel reading reached 65535 Filter four has saturated the reference channel Probable Causes The monochromator mirror grating is damaged Analog PCB intermittently failed Missed flashes or erratic flash lamp The order sorting filter wheel is jammed and not aligning the correct filter to line up with the light path or the through hole is allowing white light to pass Vo
94. during sweep mode Dark value was lt 100 during a spectral scan using 8 flashes and the number of resets saves for that wavelength during normal and rapid mode BioTek Instruments Inc Error Codes 155 General Errors Continued 0910 The measurement channel failed lt 100 or gt 20000 during filter Cont calibration or spectral scan with the flash on The measurement channel failed lt 100 during filter calibration or spectral scan with the flash off Probable Causes e If failed lt 100 Absorbance analog PCB or measurement channel analog PCB is defective Shielding of the cable between measurement channel and analog PCB is defective or disconnected Measurement channel photodetector is defective If failed gt 20000 Measurement channel photodetector is defective A faulty analog PCB or faulty internal grounding may cause internal electronic noise There may be an ambient light leak Ensure that the plate carrier door is properly closed Electrical noise may be penetrating the measurement chamber The bottom and top shrouds are part of the electrical shielding Verify that the shrouds are installed and properly fastened Order sorting filter wheel is jammed and not aligning the filter wheel to block the light path or the filter is degraded and not passing enough light energy 0911 The absorbance measurement channel Dark current value failed 0916 See Dark on the system test See
95. e Bio Cell for 1 cm wavelength readings PN 7272051 e Bio Cell adapter plate for containing up to 8 Bio Cells PN 7270512 BioTek Instruments Inc Specifications 5 Specifications Microplates Accommodates standard 6 12 24 48 96 and 384 well microplates and 60 72 and 96 well Terasaki trays Adapter PN 7330531 is required but not included Speed of Reading The actual plate read time and accuracy are dependent on the method of reading e Normal Mode is the slower of the two available modes After positioning the well over the beam the instrument waits 100 ms before taking the measurement Y Note The 100 ms delay is to allow for more complete fluid settling e Rapid Mode is faster than Normal Mode because the instrument does not wait before taking the measurement The following read times are based on a 96 well endpoint read for single or dual wavelength measurements Normal Read Mode 600 sec Rapid Read Mode 45 sec Optical Specifications range 200 to 999 nm accuracy 2nm A repeatability 0 2 nm bandpass 2 4 nm UQuant Operator s Manual 6 Chapter 1 Introduction Optical Performance 96 well flat bottom and round bottom plates single wavelength measurements Absorbance Measurement Range Accuracy Linearity Repeatability STD 0 000 to 4 000 OD 0 000 to 2 000 OD 1 0 010 OD Normal Read Mode 2 000 to 3 000 OD 3 0 010 OD Normal Re
96. e changed to 1200 or 2400 bps if necessary The Data Bits Stop Bits and Parity settings cannot be changed BioTek Instruments Inc Optional Setting Up the Serial Port for Communications 27 To change the uQuant s baud rate from the instrument keypad 1 At the Main Menu press UTIL 12 45 PM 01 23 04 DEF INE REPORT U TI L 2 Atthe SELECT UTILITY OPTION screen press SETUP ELECT UTILITY OPTION ESTS SETUP OUTPUT READ 3 Atthe EDIT SETUP INFORMATION screen press MORE to continue NFORMAT LAMBDA N FORMAT LATE 4 The SELECT BAUD RATE screen will appear showing the currently defined Baud Rate ELECT B AUD R A T E 1200 2400 9600 e Select the desired baud rate e Select VIEW to see the reader s other communication settings RS 232 SETTING 2 STOP BITS To change the uQuant s baud rate or other communications settings in Gen5 M KC4 or KCjunior refer to their respective user guides or to Appendix A Computer Control UQuant Operator s Manual 28 Chapter 3 Installation Repackaging and Shipping the pQuant D IMPORTANT Failure to properly repackage the reader increases the likelihood of damage to the instrument during shipping The shipping system was designed to stabilize the reader s mechanical mechanisms which would otherwise be free to move around during shipping If you need to ship the Quant Reader to BioTek for service or repair be sure to use the or
97. e serious error In this case turn off the instrument Upon restarting the instrument you should be able to use the keypad See Fatal Errors on page 163 for more information Contact BioTek s Technical Assistance Center for further assistance if any of error codes are displayed Refer to Chapter 1 for contact information UQuant Operator s Manual 140 Chapter 7 Troubleshooting and Error Codes General Errors General errors indicate nonfatal conditions that require attention Description and Probable Causes Abort The read or task has been aborted Abort Error The read or task has been aborted The 0101 indicates a software abort Probable Causes e User aborted read from KC4 software e User aborted from another serial interface X axis carrier in out motor did not find the opto home sensor A motor was unable to move to its home position as registered by feedback from an optical sensor Probable Causes Nylon slider bushings on the X axis rail are dirty and worn causing too much friction dirt in roller bearings causing bearings to jam Support pin on the carrier has moved preventing it from properly sitting between the two roller bearings on the bearing block Defective or broken optical sensor Defective motor controller PCB Carrier front support screws are not adjusted or are worn making the carrier uneven The support pin is no longer inserting properly into the roller bearings X axis PCB n
98. ecision balance with readability to 0 01 g e Buffer solution A or B The stock solution for Liquid Test 3 may be formulated from the chemicals listed below Solution A or by diluting a dye solution available from BioTek Solution B Solution A 10x Concentrate Phosphate Buffered Saline PBS Required Materials e Deionized water e Ingredients shown in Table 5 e B NADH powder D Nicotinamide Adenine Dinucleotide Reduced Form Sigma bulk catalog number N 8129 or preweighed 10 mg vials Sigma number 340 110 Store the B NADH powder according to the guidelines on its packaging UQuant Operator s Manual 128 Chapter 5 Instrument Qualification m 2 3 Prepare the stock buffer solution using the ingredients in Table 6 Table 6 PBS 10X Concentrate Solution Na HPO anhydrous 1 15 grams Add DI water to bring to 100 ml Mix 5 ml of the concentrated PBS solution with 45 ml of DI water Add 10 mg of the B NADH powder and mix thoroughly This is the 10x Concentrate PBS Solution Solution B Sigma PBS Required Materials Deionized water Tween 20 polyoxyethylene 20 sorbitan monolaurate Sigma P4417 tablets or equivalent B NADH Powder B Nicotinamide Adenine Dinucleotide Reduced Form Sigma bulk catalog number N 8129 or preweighed 10 mg vials Sigma number 340 110 Y Store the B NADH powder according to the guidelines on its packaging Prepare a PBS solution using the Sigm
99. ect the power cord to the external power supply 2 Locate the power supply jack on the rear of the Quant 3 Plug the rounded end of the power supply line cord into the power supply jack 4 Plug the 3 prong end of the power cord into an appropriate power receptacle Warning Power Rating The Quant power supply 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 BioTek Instruments Inc Setting Up the uQuant 21 Power Up and System Test After you have installed the Quant and connected the power supply turn on the instrument to run a system test The on off switch is located on the lower right side of the base The System Test begins with a check of the stepper motors and the analog power supplies to ensure that they have a proper input voltage level The data flash checksum motor axis and analog offset are then verified The photodetector s dark current noise and gain are checked to ensure they fall within specific pass fail criteria If an error is detected the reader will chirp and display an error code See Chapter 7 for a list of error codes If no errors are detected the reader will briefly display SYSTEM TEST PASS The power up system test does not produce a printed results report To run the test manually and obtain a printout of the system test values s
100. ed valid If the criteria are not met results are considered suspect and the message RESULTS INVALID Assay validation failed appears on results reports Up to 4 assay validation formulas can be defined Define the plate map via DEFINE gt MAP before creating assay validation formulas Examples If an assay protocol states that for the assay to be valid The mean of the negative controls must be less than 0 100 The formula is NC x 0 100 The mean of the positive controls must be greater than the mean of the negative controls The formula is PC x 5NC x UQuant Operator s Manual 68 Chapter 4 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 63 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 62 to review the results calculation structure e One transformation formula may be defined per assay e A transformation formula can be simple ex OD 2 100 to multiply all wells on the plate by 100 or more complex with the inclusion of a predefined Transformation Variable see TVAR below Simple Transformation Formul
101. eled as positive No greyzone is required For this example NC x the mean of the NC wells equals 1 000 OD The cutoff formula is NC x 0 5 The greyzone is 00 POS is selected for SAMP gt CUTOFF 00 Calls are assigned to sample wells as follows gt EQUIV if the sample equals 1 500 gt pos if the sample is greater than 1 500 gt NEG if the sample is less than 1 500 BioTek Instruments Inc Define Method Map Formula and Curve 73 2 Fora quantitative assay samples with OD values greater than the STD2 mean plus a 10 greyzone should be labeled as positive samples with OD values 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 2 000 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 EQUIV if the sample is greater than or equal to 1 800 and less than or equal to 2 200 POSifthe sample is greater than 2 200 NEGifthe sample is less than 1 800 UQuant Operator s Manual 74 Chapter 4 Operation Defining CURVE Y Note These screens are displayed on the uQuant in the order in which they appear in the assay If a closed variable is being used in the assay the entry screen is omitted To define curve fitting parameters for an assay 1 Start at the Main M
102. ell 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 BioTek Instruments Inc Liquid Testing 121 Liquid Test 1 A 96 well flat bottom microplate is required for this test Corning Costar 3590 is recommended Use a new microplate any fingerprints or scratches may cause variations in readings D IMPORTANT Before running the liquid tests ensure that the reader is not running in Rapid Mode To check this select UTIL gt READ and then cycle through the prompts until READ IN RAPID MODE is displayed Choose No for an accurate result 1 Using a freshly prepared stock solution see Solution A on page 119 or Solution B on page 120 prepare a 1 2 dilution using DI water one part stock one part DI water the resulting solution is a 1 2 dilution The concentrated stock solution should have an optical density of 2 000 or lower 2 Pipette 200 ul of the concentrated solution into Column 1 of the plate 3 Pipette 200 ul of the diluted solution into Column 2 of the plate After pipetting the diluted test solution into the microplate and before reading the plate we strongly recommend shaking the plate for four minutes This will allow any air bubbles in the solution to settle and the meniscus to stabilize If a plate Shaker is not ava
103. ement failed position verify not user serviceable Motor Y axis failed to get to the same position when moved a known number of steps from the home position and back e The optical trigger flag has moved or is loose e Foreign object in the path of the carrier Order sorting filter wheel failed positional verify Probable Causes e The optical trigger flag has moved or is loose e Filter wheel is binding against the motor gear Monochromator failed to find the zero order position white Light or positional verify The order sorting filter wheel is homed and moved to the open hole position The monochromator is moved until the optical system detects saturation It is then moved to the point where there is no saturation and then moved back to the saturation point This error is indicating the saturation did not clear or appear Probable Causes Flash lamp is missing flashes or is not flashing The optic system does not detect the saturation analog PCB Order sorting filter wheel is binding against the motor gear and not allowing the open hole to line up correctly Bearings inside the grating mirror are causing monochromator to jam UQuant Operator s Manual 144 Chapter 7 Troubleshooting and Error Codes General Errors Continued Light beam saturated too much light Air reading reached 65535 This error indicates one of the following situations e During an absorbance filter calibration or wavelength
104. ents Inc Error Codes 145 General Errors Continued 0501 Reference Channel Cont The monochromator mirror grating is damaged Analog PCB intermittently failed Missed flashes or erratic flash lamp The order sorting filter wheel is jammed and not aligning the correct filter to line up with the light path or the through hole is allowing white light to pass Voltage to the lamp has increased because the lamp power supply has failed or the motor power supply PCB sent the incorrect voltage request to the lamp power supply Light beam saturated too much light Air measurement channel reading reached 65535 This error indicates that filter two has saturated the reference channel Probable Causes Reference Channel e The monochromator mirror grating is damaged e Analog PCB intermittently failed The order sorting filter wheel is jammed and not aligning the correct filter to line up with the light path or the through hole is allowing white light to pass Missed flashes or erratic flash lamp Voltage to the lamp has increased because the lamp power supply has failed or the motor power supply PCB sent the incorrect voltage request to the lamp power supply Measurement channel light beam saturated too much light Fails if gt 3 steps of error This error indicates one of the following situations e The monochromator is unable to find the center of the white light home position or it found home in a
105. enu and select DEFINE 2 Select the assay and then press ENTER The DEFINE option screen will appear DEFINE METHOD M A P FORMULA CURVE 3 Select CURVE The definable curve fitting parameters include e Curve Fit Type e Editing of Outliers e Axis Identification e Extrapolation of Unknowns Y lt Note If a transformation formula has been defined then data used to generate a standard curve is based on the transformed data BioTek Instruments Inc Define Method Map Formula and Curve 75 Curve Fit Type The uQuant supports seven different curve fitting methods linear quadratic cubic 4 P 2 P cubic spline and point to point e Linear curve fit A simple best fit straight line is plotted using the values of the standards e Quadratic or Quad curve fit A curve fit that uses the quadratic equation ax bx c y to plot the standard s values Utilizing this curve any data point for a standard that deviates from the ideal value will not affect the entire curve e Cubic curve fit A curve fit that uses the equation ax bx ex d y to plot the standard s values This type of curve fit is affected even less than the quadratic fit when any particular standard has a poor value e 2 P LOGIT LOG 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 logist
106. er volumetric flask Table 4 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 Quant Operator s Manual 120 Chapter 5 Instrument Qualification Preparation of Stock Solution 1 Solution B Weigh out 0 092 gram of FD amp C No 5 yellow dye powder into a weigh boat Rinse the contents into a 1 liter volumetric flask Add 0 5 ml of Tween 20 or 5 ml of BioTek s wetting agent Make up to 1 liter with DI water cap and shake well This should create a solution with an absorbance of about 2 000 when using 200 ul in a flat bottom microwell The OD value 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 Required Materials BioTek OC Check Solution No 1 PN 7120779 25 ml or 7120782 125 ml Deionized water 5 ml Class A volumetric pipette 100 ml volumetric flask Preparation of Stock Solution L Pipette a 5 ml aliquot of BioTek QC Check Solution No 1 into a 100 ml volumetric flask Make up to 100 ml with DI water cap and shake well This should create a solution with an absorbance of about 2 000 when using 200 ul in a flat bottom microwell The OD value will be proportional to the volume in the w
107. es 3 and 6 Updated Hardware and Environmental Specifications page 6 Chapter 3 Installation Changed VAC to V page 20 UQuant Operator s Manual XiV Preface Revision History Continued Revision I Date 05 06 Changes General Add Geno instructions references wherever KC4 or KCjunior instructions references were included throughout the manual Changed Universal to Absorbance in Universal Test Plate or Universal Plate Test Removed the hyphen from Bio Tek BioTek Removed Universal from Universal Microplate Spectrophotomer Cover Replaced cover with new design from Marketing includes updated photo and new BioTek logo Preface Updated Notices Warnings Hazards and safety information Removed Warranty and Registration Card Chapter 1 Replaced 2 page Technical Support section with 1 page Product Support amp Service section Chapter 5 Changed chapter title from Performance Verification and IQ OQ PO Procedures to Instrument Qualification In Recommended Qualification Schedule changed PQ frequency from monthly and semiannually to monthly and quarterly and clarified criteria for running Liquid Tests 1 2 Or 3 Added Gen instructions for performing the System Test and Absorbance Plate Test Changed Sigma P 3563 packets to Sigma P4417 tablets or equivalent in required materials for Liquid Test 3 For Liquid Tests 1
108. es as well as the equations used to assign calls to samples Greyzone Entry The greyzone is a definable area around the cutoff value Samples that fall within an area defined by the greyzone ex 5 076 of the cutoff value could be considered equivocal EQUIV ENTER GREYZONE 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 Positive Negative Calls on the next page for information on how calls are assigned Quant Operator s Manual 72 Chapter 4 Operation Positive Negative Calls for Cutoff After the greyzone is defined calls for the sample wells Positive NEGative EQUIVocal must be defined Examples Select POS or NEG to select the call that will be assigned to samples ereater than the cutoff value plus the greyzone e If for example POS is selected as shown in the above screen 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 GREYZONE NEG SMP CUTOFF CUTOFF GREYZONE 1 The cutoff between negative and positive calls should be calculated as the average of the negative controls plus the OD value of 0 500 Samples ereater than the cutoff should be lab
109. f the 75 Test Solution into all wells of columns 3 and 4 150 ul of the 50 Test Solution into all wells of column 5 and 6 After pipetting the diluted test solution into the microplate and before reading the plate we strongly recommend shaking the plate for four minutes This will allow any air bubbles in the solution to settle and the meniscus to stabilize If a plate Shaker is not available wait 20 minutes after pipetting the diluted test solution before reading the plate 5 Read the microplate five times using Normal Standard Mode single wavelength at 340 nm no blanking or blank on air 6 Print the raw data or export it to an Excel spreadsheet using Gen5 KC4 M or KCjunior software 9 ey If you are exporting the data to an Excel spreadsheet perform the calculations described on the following page and keep the spreadsheet for future tests Quant Operator s Manual 130 Chapter 5 Instrument Qualification Repeatability Test 1 For each well calculate the mean and standard deviation of the five readings For each mean calculated in step 1 calculate the allowed deviation using the repeatability specification for a 96 well format of 1 0 0 005 OD Mean 0 010 0 005 For each well compare the standard deviation calculated in step 1 with the allowed deviation calculated in step 2 The standard deviation should be less than the allowed deviation For example Absorbance readings of 0 802
110. fine Method Map Formula and Curve 81 Spectral Scanning Spectral scans measure peak absorbance without using filters e To run a Spectral Scan start at the Main Menu select DEFINE and then choose assay number 00 to display the assay name LECT AS SAY NUMBER M E SPECTRAL SCAN Press ENTER to bypass the ASSAY NAME and DEFINE options and display the SELECT WAVELENGTH START screen Press ENTER to continue LECT WAVELENGTH e Enter the wavelength to START the spectral scan The valid range is 200 to 999 nm Press ENTER to continue e Select the wavelength at which to STOP the scan The valid range is the starting wavelength 1 e g 201 to 999 nm Press ENTER to continue LECT WAVELENGTH O P e Select the step increment between successive wavelengths for the spectral scan The valid range is 1 to the difference between the START and STOP wavelengths For example if START STOP STEP are set to 340 800 and 50 the uQuant will read at 340 390 440 490 and so on Press ENTER to continue LECT WAVELENGTH E P UQuant Operator s Manual 82 Chapter 4 Operation Choose the plate type Press MORE for further options Press ENTER to continue Select the well location of the scan The valid range depends on the plate geometry Press ENTER to continue ECTRAIL SCAN WELL LOCATION Choose YES to precalibrate the instrument before the scanning data is collected This procedure will col
111. gure 38 Column Report without samples uQuant Operator s Manual 194 Appendix C Report Format Bio Tek Instruments Figure 39 Panel Report Panel Date 07 31 96 Lot Time 02 24 01PM Operator Temp Plate ID COMMENTS Specinen Assay Well BIkdOD CalcOD Call RSLT Std Dev CV Notes SMPQO01 ASSAY 01 F0 006 006 ASSAY 02 B05 0 000 0 000 SMP0002 ASSAY 01 601 0 002 0 002 ASSAY 02 C05 0 007 0 007 SMP0003 ASSAY 01 HOi 0 005 0 005 ASSAY 02 005 0 004 0 004 SMP0004 ASSAY 01 A02 0 002 0 002 ASSAY 02 E05 0 012 0 012 SMP 005 ASSAY 01 802 002 002 ASSAY 02 F05 0 010 0 010 SMP0006 ASSAY 01 C02 0 005 0 005 ASSAY 02 605 001 001 SMP0007 ASSAY 01 002 001 001 ASSAY 02 H05 0 002 0 002 SMP0008 ASSAY 01 E02 0 009 0 009 ASSAY 02 A06 0 004 0 004 SMP0009 ASSAY 01 F02 0 006 0 006 ASSAY 02 806 0 007 0 007 SMP0010 ASSAY 01 602 006 006 ASSAY 02 C06 0 010 0 010 SMP0011 ASSAY 01 H02 005 005 ASSAY 02 DOG 0 004 0 004 SMP0012 ASSAY 01 A03 00 Q0 ASSAY 0 E06 0 003 0 003 SMP0013 ASSAY 01 B03 0 002 0 002 ASSAY 02 F06 001 001 SMP0014 ASSAY 01 C03 005 005 ASSAY 02 606 0 001 0 001 SMP0015 ASSAY 01 D03 002 002 Page 1 BioTek Instruments Inc 01 47PM 03 16 04 NAME READ TYPE PLATE TYPE WAVELENGTH BLANK MAP CURVE FIT TYPE EDIT STD OUTLIERS X Y AXIS TYPE EXTRAPOLATE UNKNOWNS A B C D E F G H Open Assay 04 ENDPOINT 96 405 AIR LINEAR NONE LIN NO Figure 40 Assay Detail
112. he Xenon flash bulb is filtered by the monochromator and focused into microwells and the transmitted energy is then focused on the photodetector Each flash of the bulb results in a burst of current from the photodetector This current is collected by an electronics integrator The output of the integrator is voltage This voltage is multiplied by the Gain before being digitized by the ATD analog to digital converter and passed to the microprocessor The Resets are used to get to the full scale of the ATD to ensure the best readings possible In Normal Mode 8 flashes are used per well The energy of the flashes can be accumulated on the integrator but the integrator needs to be reset before the ATD reaches full scale and overflows If the energy of a single flash is high on a given wavelength the integrator has to be reset after each flash e g 8 resets will be used for 8 flashes If the accumulated energy of two flashes does not saturate the ATD then 2 flashes will be accumulated before resetting the integrator and 4 resets will be used Two resets will be used if the energy of 4 flashes is accumulated 1 reset will be used if the energy of 8 accumulated flashes does not saturate the ATD e The Gain is used as a fine resolution multiplier to ensure the full range of the ATD is used for measurements The Signal on a given wavelength is proportional to Resets Gain When this value drops below a certain threshold the Self Test fails To moni
113. ic equation is algebraically transformed to a simpler form in which experimentally determined values are used for the responses at concentrations of zero and infinity e Cubic Spline C Spline curve fit 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 UQuant Operator s Manual 76 Chapter 4 Operation 4 Parameter Logistic or 4 P A curve fitted to the standard values which is characterized by a skewed sigmoidal S shaped plot that 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 Point to Point or PT to PT A plot that connects each standard point with a line with no averaging of the values to smooth the curve at each standard Press SOFT KEYS 1 2 3 or 4 to select the curve fit type that is displayed above the soft key Select MORE to display additional options Press ENTER to save the selection and advance to the next screen Edit Standard Outliers This screen allows you to select which method NONE or MANUAL will be used to edit Standard Outlier values After the standard curve has been calculated one or more standards can be excluded from the recalculation of the curve Any previously defined edit method
114. ids Avoid spilling liquids on the instrument fluid seepage into internal components creates a potential for shock hazard Wipe up all spills immediately Do not operate the instrument if internal components have been exposed to fluid Warning Software Quality Control The operator must follow the manufacturer s assay package insert when using the instrument s onboard software to modify software parameters and establish reading methods Failure to conduct quality control checks could result in erroneous test data Warning Reader Data Reduction Protocol The onboard assay software will flag properly defined controls when they are out of range The software will present the data with the appropriate error flags for the operator to determine control and assay validity If the reader is operated via computer control no limits are applied to the raw absorbance data All information exported via computer control must be thoroughly analyzed by the operator Warning Unspecified Use Failure to operate this equipment according to the guidelines and safeguards specified in this manual could result in a hazardous condition Quant Operator s Manual XVIII Preface Precautions The following precautions are provided to help avoid damage to the instrument A Caution Service The wQuant should be serviced by authorized service personnel Only qualified technical personnel should perform troubleshooting and service procedures on intern
115. ification Absorbance Plate Test Description Gen5 s Reader Diagnostics Utility PN 5320201 must be installed on the computer s hard drive if you wish to run the Absorbance Plate Test using Gen5 The Absorbance Test Plate is also referred to as CALPLATE on the display of the uQuant and as Universal Test Plate in KC4 and KCjunior software This test uses BioTek s 7 Filter Absorbance Test Plate PN 7260522 to confirm the mechanical alignment optical density accuracy linearity and repeatability and wavelength accuracy of the wQuant The Absorbance Plate Test compares the reader s optical density and wavelength measurements to NIST traceable values An alternate method that may be used to determine accuracy linearity repeatability and alignment is Liquid Test 2 described on page 123 The Absorbance Plate Test confirms the following e Mechanical Alignment Mechanical alignment of the plate carrier and standard microplates is confirmed by the four corner positional accuracy check e Accuracy Accuracy of the Optical Density readings the comparison of the optical density readings with those listed on the Absorbance Test Plate s Standards Certificate will confirm the accuracy of the optical density readings at specific wavelengths e Linearity Linearity of the Optical Density readings is confirmed by default if the optical density readings are accurate e Repeatability Ensures that the instrument meets i
116. iginal packing Other forms of commercially available packing are not recommended and can void the warranty If the original packing materials have been damaged or lost contact BioTek for replacement packing see Product Support amp Service in Chapter 1 for contact information Warning If the reader has been exposed to potentially hazardous material decontaminate it to minimize the risk to 1 all who come in contact with the reader during shipping handling and servicing Decontamination prior to shipping is required by U S Department of Transportation regulations Before Repackaging the Instrument 1 Decontaminate the reader before repackaging it 5ee Chapter 6 Maintenance and Decontamination for the Decontamination procedure 2 Once the instrument is clean reattach the shipping block Repackaging the Quant and Its Accessories Refer to Figures 8 through 12 when repackaging the Quant BioTek Instruments Inc Repackaging and Shipping the yQuant 29 Mounting the Shipping Block 1 Turn off the unit and unplug the power supply 2 Use the Phillips head screwdriver to remove the shipping block and mounting hardware from the rear of the unit Figure 8 and set them aside Carrier Shipping Block and Mounting Hardware Figure 8 Removing the carrier shipping block and mounting hardware Quant Operator s Manual 30 Chapter 3 Installation 3 Turn the instrument around and open the fro
117. ilable wait 20 minutes after pipetting the diluted test solution before reading the plate 4 Read the microplate five times at 405 nm using the Normal Standard Read Mode Chapter 3 single wavelength no blanking Normal plate position 5 Rotate the microplate 180 so that well A1 is now in the H12 position Read the plate five more times Turnaround plate position 6 Print the raw data or export it to an Excel spreadsheet using Gen5 KC4 or KCjunior software If you are exporting the data to your own Excel spreadsheet perform the calculations described below and on the following page and keep the spreadsheet for future tests UQuant Operator s Manual 122 Chapter 5 Instrument Qualification Repeatability Calculate the mean value for each physical well location in columns 1 and 2 for the five plates read in the Normal position and then again for the five plates read in the Turnaround position This will result in 32 mean values Perform a mathematical comparison of the mean values for each microwell in its Normal and Turnaround positions A1 H12 A2 H11 B1 G12 B2 G11 and so on To pass this test the differences in the compared mean values must be within the accuracy specification for the instrument Example comparison calculation If the mean value for well A1 in the Normal position is 1 902 where the specified accuracy is 1 0 010 OD then the expected range for the mean of the sa
118. ines Install KC4 on the computer s hard drive and register the software with BioTek Once installed start KC4 Log in if prompted Select System Readers The Reader Selection dialog will appear Scroll through the list of Available Readers and select the nQuant Click the Port button and subsequent Setup button to define the following communications parameters gt Port COMn select the COM serial port used for the RS 232 cable connection gt Transmission Speed 2400 4800 or 9600 default must match the baud rate on the reader Data Bits 8 parity No Stop Bits 2 Although the transmission speed baud rate may be changed to 2400 or 4800 BioTek recommends keeping the default rate of 9600 Click OK to close the Port Setup dialog then OK again at the Port Selection dialog BioTek Instruments Inc Controlling the Reader with KC4 171 9 Click Filters Wavelengths then Get Wavelengths to align KC4 s wavelengths table with the reader s internal table Click OK to return to the Reader Selection dialog 10 Click the Current Reader button to attempt to establish communications with the reader using the currently defined communication parameters 11 If the test passes click Close to save the settings and return to the main screen If the test fails KC4 will provide appropriate instructions for resolving any problems See also the Problems section below Problems
119. ing and Inspecting the uQuant 19 8 Attach the shipping block and mounting hardware to the rear of the unit as shown in Figure 6 Carrier Shipping Block and Mounting Hardware Figure 6 Attaching the shipping block and mounting hardware to the rear panel UQuant Operator s Manual 20 Chapter 3 Installation Setting Up the Quant Operating Environment The uQuant is designed to operate optimally when installed on a level surface in an area where ambient temperatures remain between 18 C 64 4 F and 40 C 104 F The reader is sensitive to extreme environmental conditions and these conditions should be avoided Excessive humidity Condensation directly on the sensitive electronic circuits can cause the instrument to fail internal self checks The specified humidity range for this instrument is from 10 to 85 noncondensing Excessive ambient light Bright sunlight or strong incandescent light may affect the instrument s optics and readings reducing its linear performance range Dust Optical density readings may be affected by extraneous particles such as dust in the microplate wells A clean work area is essential to ensure accurate readings Electrical Connections MN Caution Power Supply Only use the specified power supply to ensure proper operation of the unit The Quant has a universal 24 VDC 2 A power supply that functions from 100 to 240 V 10 50 to 60 Hz without external switching 1 Conn
120. ioTek Instruments Inc Safety Symbols Safety Symbols XXI Some of the following symbols will appear on the instrument EN e Alternating current Courant alternatif Wechselstrom Corriente alterna Corrente alternata Direct current Courant continu Gleichstrom Corriente continua Corrente continua Both direct and alternating current Courant continu et courant alternatif Gleich und Wechselstrom Corriente continua y corriente alterna Corrente continua e corrente alternata Earth ground terminal Borne de terre Erde Betriebserde Borne de tierra Terra di funzionamento Protective conductor terminal Borne de terre de protection Schutzleiteranschluss Borne de tierra de protecci n Terra di protezione On Supply Marche alimentation Ein Verbindung mit dem Netz Conectado Chiuso Off Supply Arr t alimentation Aus Trennung vom Netz Desconectado Aperto sconnessione dalla rete di alimentazione UQuant Operator s Manual xxii Preface A e A an o A H Warning risk of electric shock Attention risque de choc lectrique Gefahrliche elektrische schlag Precauci n riesgo de sacudida el ctrica Attenzione rischio di scossa elettrica Warning risk of crushing or pinching Attention risque d crasement et pincement Warnen Gefahr des Zerquetschens und Klemmen Precauci n riesgo del machacamiento y sejeci n Attenzione rischio di schiacciare ed intrappolarsi Warning
121. is displayed ED IT S T D OUTLIERS MANUAL NONE MANUAL Select NONE to suppress the EDIT STANDARD OUTLIERS capability for this assay Choose MANUAL to enable the capability gt If AVERAGE STANDARDS is set to NO the individual standard replicates are available for editing If set to YES the standard groups are available for editing gt After the assay is run and reports are generated press REPORT from the Main Menu Press RESULT select the assay and then press ENTER The EDIT STD OUTLIERS YES NO prompt will appear See Editing Standard Outliers on page 89 for further instructions BioTek Instruments Inc Define Method Map Formula and Curve 77 X Y Axis Type lt Note Linear only LIN LIN axes are recommended when using the Cubic Spline fit After the curve fit type is selected select the X Y Axis Type e Select 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 This screen allows you to choose whether to extrapolate the curve to evaluate samples outside of the absorbance range defined by the standards EXTRA POLAT SE UNKNOWNS YES N O Select YES to enable extrapolation otherwise select NO On the printed reports extrapolated concentrations RSLT values are surrounded by lt gt e g lt 44 425 gt D If extrapolation is chose
122. ition Next compare the mean values for the H1 well to the same well in the A12 position 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 OD then the expected range for the mean of the same well in the H12 position is 1 873 to 1 931 OD 1 902 1 0 019 0 010 0 029 which is added and subtracted from 1 902 for the range If the four corner wells are within the accuracy range the reader is in alignment Accuracy Specifications For comparison in this test the following accuracy specifications are applied using Normal Standard Mode and a 96 well microplate 1 0 010 OD from 0 000 to 2 000 OD t 376 0 010 OD from 2 000 OD to 3 000 OD BioTek Instruments Inc Liquid Testing 127 Liquid Test 3 Liquid Test 3 is an optional test offered for sites that must have proof of linearity at wavelengths lower than those attainable with the Absorbance Test Plate This test verifies operation of the uQuant M at 340 nm and is optional because the reader has good front end linearity throughout its wavelength range Required Materials e 340 nm filter installed in the reader e New 96 well flat bottom microplates Corning Costar 3590 is recommended e Calibrated hand pipette s e Beakers and graduated cylinder e Pr
123. its accompanying Standards Certificate The six wavelengths can be set at any time The default values can be changed via Gen5 KC4 or KCjunior refer to the Gen5 Getting Started Guide or Help system or to the User s Guides to KC4 or KCjunior To check or change the software wavelength from the instrument keypad 1 Power up the Quant 2 From the Main Menu press UTIL The UTILITIES menu appears 45 PM 01 23 04 N E REPORT UTI L LITY OPTION P OUTPUT READ 3 From the SELECT UTILITY OPTION menu press SETUP The EDIT SETUP INFORMATION screen is displayed SETUP INFORMATION T IME LAMBDA MORE 4 From this menu press LAMBDA The wavelength will be displayed Press ENTER to continue BioTek Instruments Inc Setting Up the uQuant 23 6 To change the wavelength number use the numeric keypad to enter a number at the cursor location The cursor will automatically advance to the next editable field Press ENTER to save the entry and move to the next wavelength on the table ENTER LAMBDAHd 1 WAVELENGTH Note Any wavelength value between 200 and 999 nm may be entered in 1 nm increments 7 When the last wavelength has been entered the software exits the routine and displays the following screen SETUP INFORMATION T IME LAMBDA MORE 8 Press the Main Menu key 9 Perform a System Test to calibrate the system for the new lambda values SYSTEM TEST PASS will be displaye
124. l 26 Chapter 3 Installation Optional Setting Up the Serial Port for Communications Before serial communication can be initiated between the Quant and another device such as a host PC running BioTek s Gen5 KC4 or KCjunior software the communication parameters must match between the devices The uQuant has a 25 pin serial RS 232 port located on the rear panel of the instrument The serial port allows the reader to communicate with a computer using standard communications software and or RS 232 protocols The serial port also allows field upgrades of the Quant software Appendix A contains information on required protocols for computer control of the reader Attaching the Cable 1 Power down the computer and the Quant 2 Connect the appropriate serial cable to both machines The serial port on the reader is a DTE configuration with a 25 pin pin male D sub connector Power up the reader and the computer 4 Ensure that the Quant and the computer are operating with the same communications settings Installing the PC Software Install Gen5 KC4 or KCjunior software on the computer s hard drive Refer to the Gen5 Getting Started Guide or Help system or to the User s Guides for KC4 or KGjunior for installation instructions Setting Communication Parameters The reader s default communication parameters are e 9600 Baud Rate e 6 Data Bits e 2 Stop Bits e No Parity The baud rate can b
125. lect the new dark and air blank values Otherwise choose NO and press ENTER to return to the Main Menu CALIBRATE O BioTek Instruments Inc Reading a Microplate 83 Reading a Microplate Use the READ option found at the Main Menu to select an assay to run define any required run time options and then begin a plate read Y Note Before reading a plate ensure that the reporting options are set correctly under UTIL OUTPUT To read a plate 1 Start at the Main Menu and select READ The SELECT ASSAY NUMBER screen will appear 2 Select an assay and then press ENTER See Selecting an Assay to Run on the next page for detailed instructions 3 If required enter the number of Samples Plate ID and or Sample ID Note The options to present these screens are configurable by selecting UTIL gt READ See Read Options in Chapter 3 for more information 4 Place the plate in the carrier and then press the READ key to continue Y Quant Operator s Manual 84 Chapter 4 Operation Selecting an Assay to Run To select an assay to run start at the Main Menu and select READ The SELECT ASSAY NUMBER screen will appear LECT AS SAY NUMBER M E H B S AG 1 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 e Press ENTER to select the assay and continue Run Time Prompts After the
126. llowing situations The reference channel failed lt 100 during optic test with the flash on The reference channel failed lt 100 during a read or blank read not in sweep mode with the flash off The reference channel failed lt 100 or the Dark value has changed more than 10 from the last self test data during a read or blank read with the flash on Probable Causes Absorbance analog PCB or reference channel analog PCB is defective Cable between reference channel and analog PCB is defective or disconnected Reference channel photodetector is defective or the optic spray is damaged A faulty analog PCB or faulty internal grounding may cause internal electronic noise There may be an ambient light leak Ensure that the plate carrier door is properly closed Electrical noise may be penetrating the measurement chamber The bottom and top shrouds are part of the electrical shielding Verify that the shrouds are installed and properly fastened Order sorting filter wheel is jammed and not aligning the filter wheel to block the light or the filter is degraded and not passing The absorbance measurement channel Dark current value failed See Dark on the system test See criteria in text below This error indicates one of the following situations e The measurement channel failed during a read or spectral scan for one of the following reasons Dark value was 100 during a spectral scan using 8 flashes and 8 resets
127. lt rate of 9600 for the pQuant and may not be changed 6 Click Test Comm Gen5 will attempt to communicate with the reader BioTek Instruments Inc Reading a 384 Well Plate in Gen5 179 gt If you receive The Reader is communicating message click OK and then click OK again to save the settings Click Close at the Reader Configuration dialog to return to the main screen gt Ifthe test is not successful and you receive an error message refer to the Troubleshooting section of the Help system 7 Atthe main screen select File New Experiment The New Experiment dialog will appear Mew Experiment lx Select a Protocol Madified 2 Default Protocol Cancel Browse Help Figure 22 New Experiment dialog 8 Click Default Protocol then click OK Gen5 will open the Experiment workspace which includes the Protocol menu and Plate screen Ge Gens Experiment1 File Plate Protocol Window System Help SAA PONE HSE t Plate 1 E E Protocol Matrix Plate 1 02 Audit Trail Edit Mask Figure 23 Experiment workspace UQuant Operator s Manual 180 Appendix B Using 384 Well Plate Geometry 9 Select Plate Read or click the Read Plate icon to open the Procedure dialog Use the drop down arrow in Plate Type to select a 384 well plate E Procedure uQuant Comi 4 X Add Step Advanced Options Read
128. ltage to the lamp has increased because the lamp power supply has failed or the motor power supply PCB sent the incorrect voltage request to the lamp power supply BioTek Instruments Inc Error Codes 147 General Errors Continued Light beam saturated too much light Air reading reached 65535 Filter five has saturated the reference channel Probable Causes The monochromator mirror grating is damaged Analog PCB intermittently failed Missed flashes or erratic flash lamp The order sorting filter wheel is jammed and not aligning the correct filter to line up with the light path or the through hole is allowing white light to pass Voltage to the lamp has increased because the lamp power supply has failed or the motor power supply PCB sent the incorrect voltage request to the lamp power supply Light beam saturated too much light Air measurement channel reading reached 65535 Filter six has saturated the reference channel Probable Causes The monochromator mirror grating is damaged Analog PCB intermittently failed Missed flashes or erratic flash lamp The order sorting filter wheel is jammed and not aligning the correct filter to line up with the light path or the through hole is allowing white light to pass Voltage to the lamp has increased because the lamp power supply has failed or the motor power supply PCB sent the incorrect voltage request to the lamp power supply UQuant Operator
129. lyzed for each concentration The well that shows the most variation for each concentration is selected for data reduction 2 For each mean below 2 000 OD calculate the allowed deviation using the repeatability specification for a 96 well format of 1 0 0 005 OD If above 2 000 OD apply the 3 0 005 specification 3 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 OD which is the allowable deviation Since the standard deviation is less than this value the reader meets the test criteria Repeatability Specifications 1 0 005 OD from 0 000 to 2 000 OD 3 0 005 OD from 2 000 to 3 000 OD Quant Operator s Manual 126 Chapter 5 Instrument Qualification Alignment Test 1 Using the prepared plate conduct a turnaround test by reading the plate with the A1 well in the H12 position five times This test results in values for the four corner wells that can be used to determine alignment Calculate the means of wells A1 and H1 in the normal plate position data is from Linearity Test and in the turnaround position from Step 1 above Compare the mean reading for well A1 to its mean reading when in the H12 pos
130. me well in its Turnaround H12 position is 1 873 to 1 931 OD 1 902 0 010 0 010 0 029 1 902 0 029 1 873 1 902 0 029 1 931 Accuracy Specification For comparison in this test the following accuracy specifications are applied using Normal Standard Mode and a 96 well microplate 1 0 010 OD from 0 000 to 2 000 OD 3 0 010 OD from 2 000 to 3 000 OD BioTek Instruments Inc Liquid Testing 123 Liquid Test 2 The recommended method of testing the instrument performance is to use the Absorbance Test Plate to confirm alignment repeatability and accuracy which will also confirm linearity If a Test Plate is not available Liquid Test 2 can be used for these tests Required Materials e New 96 well flat bottom microplates Corning Costar 3590 is recommended e Ten test tubes numbered consecutively stored in a rack e Calibrated hand pipette Class A volumetric pipette recommended e Stock solution A or B these are the same solutions as for Liquid Test 1 Preparation of Dilutions 1 Set up a rack containing 10 tubes numbered consecutively 2 If you have not already done so prepare concentrated stock Test Solution A or B Refer to Table 5 when executing steps 3 and 4 Table 5 Test Tube Dilutions s je 7 s 9 10 IEEE Tube Number Volume of Concentrated E Solution ml Absorbance expected if concentrated solution is 2 0 at 200 ul KERE Volume of 0 05 Tween 4 10 1
131. mits 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 BioTek Instruments Inc Recommended Qualification Schedule 99 Recommended Qualification Schedule The schedule shown in Table 2 defines the factory recommended intervals for performance testing for a microplate reader used for one shift seven days a week Y 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 2 Recommended Qualification Schedule Installation Operational Performance Qualification Qualification Qualification Initially Initially Annually Monthly Quarterly System Self Test p 101 Checksum Test p 107 AAA Absorbance Plate Test p 108 Liquid Test 1 p 121 i NENNEN Liquid Test 2 p 123 Js NENNEN Liquid Test 3 p 127 Optional for 340 nm If you 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
132. mmended Qualification Schedule seen 99 Qualification PrOCediuF ess scu d aD dea n ad ac Rain ea d cete cis un bae ca E CR b 100 SVSEem Tests ira ia oia 101 Checksum TES Enirar bio ato ei 107 ABSOrDalnce Plate Tests cisci iio loli 108 EAQUIG TESTING is 117 REPS DINGY sai ai a 118 UQuant Operator s Manual vi Preface AMAN ea aa e aa 118 Stock Solution F rmUlat OR serna aaa 119 quid ROSE Traan aaa aa aaa 121 quid ROSE Z raaa iio 123 Haud Testo la as 127 PFOCCGUES danita 129 MaintenNaN CG EET 132 Routine Cleaning Procedure scaricare 132 PUDO iaa laa 132 TOools and SUPPIES rial 133 Procedu esnea o ae eh at beta ret tis eas tee betas 133 Decontamination Procedure as 134 TOOIS ANG SUPDINCS aaa 135 PFOCCGUE a do 135 OVCIVICW Meet TERTII PT 138 System Test Descriptio nasa aia C ES 138 Glossary of TENSA a 138 ALO AA COGS Co REP nn A aa 139 E al Ei Ol EIE Lo A 140 Fatal ENOPS pores ees rot docsters lid aia 163 Controlling the Quant with Gen5 KC4 or KCjunior SOTCW Al GC ii ii a a LOD OV CIVICW O 166 Controlling the Reader with Gen5 ce cececccsesseeeeesssecneereecseeuseneeensegnaanye 167 Seng UD GON Sicvevvwes cea E QU T 167 PFODIG APP O 168 Getting Started With GON Sian 168 Controlling the Reader with KC4 cceccesssesnnereeecccuseeeeesseunnareesseeueenes 170 Sewing UD QU cia os 170 BioTek Instruments Inc Contents Vii muelsl lrcecer A aaa a a a a aaa 171
133. modifying an assay automatically saves the current settings Cycle through available options within a screen For example press Options within the Select Assay Number screen to cycle through the names of the onboard assays ENTER Pressing ENTER generally saves the current screen settings and advances to the next screen in a series Previous Pressing Previous Screen generally saves the current screen settings Screen and returns control to the screen most previously viewed CLEAR Press CLEAR to reset a numeric value to 0 or to clear all characters when editing an assay name Tip Press Shift Clear at the Map Generation screen to clear a previously defined manual map Move the cursor to the left in data entry screens Move the cursor to the right in data entry screens Initiate a plate read Halt the read currently in progress OOH HAH LF UQuant Operator s Manual 38 Chapter 4 Operation Overview IMPORTANT Do not turn on the instrument until the carrier shipping block has been removed The uQuant features a 25 pad keypad and a 2 line x 24 character LCD display allowing you to access the reader s program menus and print test results The reader s bidirectional serial port allows computer control of the instrument and provides the means for downloading additional assay definition files to the instrument This chapter describes the operation of the open configurable assays onboard the uQuant Y The
134. n the assay Wells A01 through H01 are valid for ACROSS mapping A01 through H12 are valid for DOWN Repeat this process for each assay within the panel Remain aware of the total number of controls standards and blanks that were originally mapped in each 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 The mapping for Assay 8 can begin in well B04 or any well other than A01 to B03 The reader will chirp if you try to map into a well that is already assigned for use with the Panel The mapping for Assay 22 may begin at the next available well location after Assay 8 mapping is complete 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 The Panel Assay results are sorted by sample unless a custom assay has been programmed by BioTek lt Note The interpretation of Results reports for each assay in the Panel will print first and then the Sample results will print BioTek Instruments Inc De
135. n 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 such All calculated results for an Ambiguous curve should be considered unreliable UQuant Operator s Manual 78 Chapter 4 Operation Panel Assays A Panel assay is a collection of up to 8 assays to be run on one plate e The most common reason to use a Panel assay is for confirmatory tests based on a screening test in clinical applications e Only one panel can be defined on the reader at any time e The assays specified within the Panel must be predefined in any of the assay positions 1 55 e The assays specified within the Panel must all use the Endpoint read method e The assays specified within the Panel must all read at the same wavelength s e Any curve fit type formulas or standard concentrations previously defined for each assay will be used when the assay is selected for a Panel e The type and number of controls blanks standards and replicates in the assays chosen for the Panel will be copied into the Panel definition Map or assay parameters must first be changed in the predefined assay before they can change in the Panel Tip For use with mapping the Panel consider printing a
136. n the number of empty wells remaining on the plate the reader will chirp 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 D Use caution when creating multiple Plate IDs The reader does not warn you that you about to exceed the maximum of 8 plate IDs stored in memory If a ninth Plate ID is added it will overwrite the first Plate ID stored in memory If the optional internal bar code scanner is installed the reader will automatically scan the plate bar code label and use this as the Plate ID UQuant Operator s Manual 86 Chapter 4 Operation Use up to 10 alphanumeric characters See pages 36 and 33 for instructions on using the keypad Press ENTER to advance to the ENTER SAMPLE ID screen Enter Sample ID If the ENTER SAMPLE ID prompt is presented enter a starting sample identification number e The valid entry range is from 0001 to 9999 e The software will automatically increment each subsequent sample identification number by 1 e The sample IDs will be assigned according to the plate map defined in the assay Prompts for Well Location If the assay specifies manual plate mapping and if PROMPT FOR SAMPLE COUNT is set to YES under UTIL gt READ sample well locations can be defined at run time SAMPLE LOCATION
137. nd press ENTER Enter STD1 as the TVAR formula Press ENTER The formula selection screen is displayed Choose TRANS Enter the formula OD TVAR 100 using the MATH OTHER MAP and FUNCTION keys IVAR is available as a MAP option BioTek Instruments Inc Define Method Map Formula and Curve 71 Cutoff Formulas A cutoff formula calculates a cutoff value that is used for classifying samples See Formula Type on page 63 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 e One cutoff formula may be defined per assay e If Transformation Formulas are defined cutoffs are based on the transformed results Refer to Defining Formula on page 62 for the order in which formulas are processed e A cutoff formula can consist of a simple numeric value 1 500 a well identifier PC to indicate the criterion for each of the PC replicates or PC x to indicate the average of the Positive Control replicates or a formula combining the two NC x 0 050 e A greyzone around the cutoff value can be defined to indicate equivocal or indeterminate results e Donotuse the lt or gt mathematical symbols in a cutoff formula Tip Choose to print a Column Report to see the greyzone and cutoff valu
138. nt door While holding the carrier remove the two black dress screws from the carrier shipping block mounting holes inside the carrier and set them aside 4 Install the carrier shipping block and mounting hardware that were removed from the rear panel Figure 9 IMPORTANT Do NOT remove the two black dress screws that secure the carrier sliding block to the carrier Remove Black Dress Screws and replace with carrier shipping block and mounting hardware from rear panel Black Dress Screws Do NOT Remove Carrier Sliding Block Figure 9 Removing the black dress screws from the carrier shipping block mounting holes and attaching them to the rear panel BioTek Instruments Inc Repackaging and Shipping the uQuant 31 5 Attach the two black dress screws that were removed from the carrier shipping block mounting holes to the rear panel Figure 10 Black Dress Screws Gra Figure 10 Attaching the black dress screws to the rear panel UQuant Operator s Manual 32 Chapter 3 Installation Packing the Reader and Accessories 1 Insert bottom foam end cap all the way into the bottom of the box Note the orientation of the foam to the box Figure 11 2 Put the instrument inside the 2 mil poly bag and place it inside the bottom end cap 3 Put the power supply inside the 8 1 2 x 11 bubble bag Place the power supply inside the pocket in the box as shown 26 x 32 2 Mil Poly Bag Figure 11
139. ntact the BioTek Service Department Do not soak the keypad this will cause damage D Important Turn off and unplug the instrument for all decontamination operations 1 Turn off and unplug the instrument 2 Prepare an aqueous solution of 0 5 sodium hypochlorite NaClO or bleach As an alternative 7076 isopropyl alcohol may be used if the effects of bleach are a concern gt Besureto check the percent NaClO of the bleach you are using this information is printed on the side of the bottle Commercial bleach is typically 1076 NaCIO if this is the case prepare a 1 20 dilution Household bleach is typically 5 NaClO if this is the case prepare a 1 10 dilution UQuant Operator s Manual 136 Chapter 6 Maintenance and Decontamination 3 Moisten a cloth with the bleach solution or alcohol Do not soak the cloth 4 Wipe the keypad do not soak Wipe again with a clean cloth moistened with deionized or distilled water Dry immediately with a clean dry cloth 5 Wipe the plate carrier and all exposed surfaces of the instrument 6 Wait 20 minutes Moisten a cloth with deionized or distilled water and wipe all surfaces of the instrument that have been cleaned with the bleach solution or alcohol 7 Useaclean dry cloth to dry all wet surfaces Discard the used gloves and cloths using a Biohazard trash bag and an approved Biohazard container BioTek Instruments Inc Chapter 7 Troubleshooting and Er
140. ntaminated with a cloth moistened not soaked with water or water and a mild detergent BioTek Instruments Inc Routine Cleaning Procedure 133 Tools and Supplies e Mild detergent e Deionized or distilled water e Clean cotton cloths Procedure 1 Turn off and disconnect the instrument from the power supply 2 Moisten a clean cotton cloth with water or with water and the mild detergent Do not soak the cloth Wipe the plate carrier and all exposed surfaces of the instrument 4 If detergent was used wipe all surfaces with a cloth moistened with water 5 Useaclean dry cloth to dry all wet surfaces UQuant Operator s Manual 134 Chapter 6 Maintenance and Decontamination Decontamination Procedure Any laboratory instrument that has been used for research or clinical analysis is considered a biohazard and requires decontamination prior to handling Decontamination minimizes the risk to all who come into contact with the instrument during shipping handling and servicing Decontamination is required by the U S Department of Transportation regulations Persons performing the decontamination process must be familiar with the basic setup and operation of the instrument BioTek Instruments Inc recommends the use of the following decontamination solutions and methods based on our knowledge of the instrument and recommendations of the Centers for Disease Control and Prevention CDC Neither BioTek nor the
141. nted and or uploaded to controlling software on a host PC The uQuant is backed by a superior support staff If the instrument ever fails to work perfectly please contact BioTek s Technical Assistance Center refer to page 7 for contact information Quality Control It is considered good laboratory practice to run laboratory samples according to instructions and specific recommendations included in the package insert for the test to be conducted Failure to conduct Quality Control checks could result in erroneous test data BioTek Instruments Inc Hardware Features 3 Hardware Features e Xenon flash light source e Single channel measurement system with reference channel e Monochromator with wavelength range of 200 999 nm e 2 line x 24 character LCD display e Membrane keypad with 25 alphanumeric keys e X Y carrier movement e Capability of reading 6 12 24 48 96 and 384 well microplates e External 24 volt power supply compatible with 100 240 V 50 60 Hz e One serial COM port 25 pin male connector e One parallel port 25 pin female connector Software Features e An easy to use menu driven interface e Endpoint calculations e Curve Fitting with 4 parameter cubic quadratic linear 2 P cubic spline and point to point methods e Transformation and Formula calculations for more complex mathematical operations including validations e Assays that can be programmed into memory and recalled instantly e Up
142. nts menu options as they appear on the instrument s display Note Bold text is primarily used for emphasis This icon calls attention to important information uQuant Operator s Manual Xii Preface Revision History Revision Date Changes A 7 98 Release to Production B 7 98 Changed A accuracy from 3 nm to 2 nm pages 1 6 and 4 16 C 9 98 Added reference to 384 well microplate in Specifications page 1 6 D 2 99 Revised Appendix B Computer Control Added Appendix E Error Codes Revised compatible printer in Chapter 2 E 7 02 Added references to new microplate geometries 6 and 12 plate throughout manual Deleted section on Reuse of Standard Curves Chapter 3 Revised Preface Chapters 2 and 4 and Appendices B and E Edited format and text as necessary E 10 03 Updated Notices with current contact information Updated Intended Use to distinguish between European Union and all other jurisdictions Updated Warranty to include Bio Tek s current warranty statement Clarified bleach dilution solutions for Decontamination G 5 04 General Restructured layout of manual Edited and reformatted text according to new template Modified appearance of display screens Changed Abs to OD throughout Preface Updated information in Notices page ii Updated contact information page ix Expanded the Intended Use Statement page xv Updated all user safety sections pages xvi xxi Chapter
143. ocations on the plate that are undefined The maximum number of control groups is 8 BioTek Instruments Inc Define Method Map Formula and Curve 59 Control Type After defining the number of controls for this assay select the types of controls to use 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 screen entry screen is presented if the number of control groups is greater than 0 EN TER NUMBER OF REPLICATES O F P C e The well ID associated with Control 1 appears first Press ENTER to advance to the next control e Use the numeric keys to enter a value for Number of Control Replicates UQuant Operator s Manual 60 Chapter 4 Operation 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 Location of Controls Use this option to enter the location of controls in the assay CONTROL 1 LOCATION TY PE PC RE P 1 A02 Y The displayed location field can only be edited if manual ma
144. on 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 STD5 The assay data reduction states Subtract the mean of CTL1 from the mean of the NC Subtract the difference from all ODs on the plate Divide the result of the above by the mean of the NC less the mean of CTL1 and then multiply by 100 Quant Operator s Manual 70 Chapter 4 Operation 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 gt At the SCOPE VARIABLE selection screen choose OD and press ENTER Enter the formula NC x CTL1 x by using the MATH OTHER MAP and FUNCTION keys Press ENTER The formula definition screen is displayed Choose TRANS Enter the formula OD TVAR TVAR 100 using the MATH OTHER MAP and FUNCTION keys TVAR is available as MAP option Example In the case of competitive reactions converting absorbance data to percent B Bo can be OD STD1 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 a
145. ontinue to scroll through the six available wavelength positions pressing ENTER between each When all six wavelengths have been defined or skipped the software will automatically return to the EDIT SETUP INFORMATION menu Specifying Data Output and Reporting Options Plate data can be sent to an attached printer or external computer e The onboard software provides several different options for report format and content e Data sent to an external computer has no data reduction applied to it with the exception of dual wavelength subtraction if defined in the assay Any reporting options selected through the onboard software have no effect on serial output To specify data output and reporting options start at the Main Menu and select UTIL gt OUTPUT The REPORT OUTPUT screen will appear RE PORT OUTPUT BOTH PRINT COMPUTER BOTH gt 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 UQuant Operator s Manual 94 Chapter 4 Operation Note These options have no effect on data output if the instrument is being controlled by software such as Gen5 KC4 or KCjunior running on a host PC gt Press ENTER to continue The SELECT PRINTER screen will appear SELECT PRINTER EPS ON H P gt The Quant supports printers using either HP s PCL3 language such as the HP DeskJet series
146. ook longer than kinetic interval Probable Causes e User defined interval is incorrect No absorbance A D ready transition This error indicates that the A D chip on the analog PCB is not ready for input from the photodiodes BioTek Instruments Inc Error Codes 163 Fatal Errors Fatal Errors indicate conditions that require immediate attention If a fatal error is displayed contact BioTek s Technical Assistance Center for further instructions Refer to Chapter 1 for contact information Description and Probable Causes Task control block not available Read already in progress Motor not available This error indicates that a motor is not available but it does not identify which motor was requested A600 Data flash write timed out A700 Data flash readback did not match write A800 Code flash write timed out A900 Memory allocation heap corrupted UQuant Operator s Manual 164 Chapter 7 Troubleshooting and Error Codes BioTek Instruments Inc Appendix A Controlling the Quant with Gen5 KC4 or KCjunior Software The uQuant can be controlled either from BioTek s Gen5 KCjunior or KC4 PC software This Appendix provides instructions for programming the computer to control the instrument eR ia aa 166 Controlling the Reader with Geno 0 cccssseeeceeecenseeeeessseunnnttessseans 167 SENG UD BES oe ea do el A 167 POD ISS riadas 168 Getting Started with Genb
147. or Epson s LQ language For the latest list of compatible printers consult the BioTek Web site www biotek com or call BioTek Instruments Technical Assistance Center refer to Chapter 1 for contact information gt Select EPSON or HP as appropriate gt Press ENTER to continue The REPORT TYPE screen will appear COLUMN MATRIX BOTH Thecurrently 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 ex 8 x 12 matrix or BOTH See Appendix C for examples of Reports Press ENTER to continue The SAMPLES IN COLUMN REPORT screen will appear Select YES to print results for all wells on the plate including samples Select NO to limit the results information to blanks controls and standards gt Press ENTER to continue The PRINT CURVE FIT screen will appear PRINT CURVE FIT N O BioTek Instruments Inc Utility 95 gt Select YES to print the standard curve only applies to quantitative assays gt Press ENTER to return to the SELECT UTILITY OPTION screen Selecting Read Options At plate read time the software can be configured to present the user with a series of prompts to enter information such as Plate ID Sample ID or Sample Count To specify various read time options start at the Main Menu and select UTIL 3 READ The PROMPT FOR PLATE ID sc
148. oreader The following calibration data has been recorded by a N I S T traceable spectrophotometer WAVELENGTH nm wu oe MM 1 537 1 272 Set 2453 Serial 161259 Figure 17 Sample Standards Certificate for the Absorbance Test Plate Before the Absorbance Plate Test can be performed the standard OD values and the peak wavelength value s must be entered via the keypad or via Gen5 M KC4 M or KCjunior software This only has to be done once Instructions for entering the Absorbance Test Plate data and for running the test are provided on the following pages UQuant Operator s Manual 110 Chapter 5 Instrument Qualification Entering the Absorbance Test Plate Data Use the Standards Certificate and Peak Wavelength Certificate included with the test plate when performing the instructions below and on the following page To enter the test plate data into the reader from the instrument keypad 1 Start at the Main Menu and select UTIL gt SELECT UTILITY OPTION gt SETUP The EDIT SETUP INFORMATION menu will appear SETUP INFORMATION T IME LAMBDA MORE 2 Press MORE to display the second EDIT SETUP menu Select CALPLATE to access the CALIBRATION LAMBDA menu 3 Using the Standards Certificate packaged with the test plate select the filter wavelength SETUP INFORMATION CALPLATE MOR E ALIBRATION LAMBODA 4 0 5 45 0 49 0 4 Press ENTER The WELL LOCATION CALIBRATION VALUE screen appea
149. ot adjusted far enough to the right This will not allow the flag to enter the opto sensor enough to trip the sensor Loosen the two screws and slide the PCB to the right and retighten Run the carrier autocal Carrier not able to move into read chamber An object may be obstructing the carrier s path BioTek Instruments Inc Error Codes 141 General Errors Continued Y axis carrier left right motor did not find the home sensor A motor was not able to move to its home position as registered by feedback from an optical sensor Probable Causes Bearings on the Y axis rails are dirty and worn and causing too much friction Defective or broken optical sensor Defective motor controller PCB or cable Carrier not able to move into read chamber Check the carrier s path for obstructions Order sorting filter wheel did not home Probable Causes Filter wheel is not tight and can wobble Filter wheel is too close to the motor gear and is binding Defective or broken optical sensor Defective motor motor controller PCB or cable Saturation transition failed in the X axis movement light beam never found During the X axis movement the light beam saturation transition max light to no light was never found during autocalibration Probable Causes Dirty rail and nylon bushings or bearings causing the carrier to jam Autocal jig is not in the carrier Order sorting filter wheel is jammed and unable
150. ow Press ENTER to save the selection and continue Replication Direction This option allows you to specify how replicates are mapped on the plate RE P DIRECTION ACROSS DOWN ACROSS e Select DOWN to map the replicates down the column following the direction of the map listing e Select ACROSS to map the replicates across in a paired format As an example two replicates can be placed in A1 and A2 wells The third replicate would follow in B1 The next standard control or sample would follow in B2 e Press ENTER to save the selection and continue e Examples of mapping directions are shown on the next page UQuant Operator s Manual 50 Chapter 4 Operation Examples of Mapping Directions Map Direction DOWN Rep Direction DOWN Bo dg 12 5 45 T5 de 7 qe 9 0 E 42 A SIDL SID 8MP P B SIDL SID MP y ISID2 PC SMP y D SI2 FPC y P LIE SIDA NE py 1 A E EOS E E cp Lo F sm3pNne J j J j j pp jp G sra se j qp pp P LEE SIDA MEL 1L 1 1 Jg p SY Y v jy Map Direction ACROSS Rep Direction ACROSS Le NN NEN TN TRIN E ANN 7 8 yg 10 ji am SMP SMP SMP SMP_ SMP SMP MEN ETS MEN NE A STD e Nc Nc NUN Lu LL E So T S T S T S f S NNNM AS E AE E Gh y a NN Map Direction DOWN Rep Direction ACROSS EE EP e A stbi srbi
151. pping was selected see page 49 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 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 gt READ gt PROMPT FOR SAMPLE COUNT is set to YES See Read Options in Chapter 3 for more information e Use the numeric keys to enter the number of sample groups on the plate e The range is 0 to the number of undefined well locations remaining on the plate For example if there are no controls blanks or standards defined on a 96 well plate the maximum number of samples is 96 and the minimum number of samples is 1 BioTek Instruments Inc Define Method Map Formula and Curve 61 Number of Sample Replicates After the number of sample groups is specified the NUMBER OF SAMPLE REPLICATES entry screen is presented TER NUMBER OF M P LE REPLICATES e Use the numeric keys to enter the NUMBER OF SAMPLE REPLICATES e The valid 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 MANUAL MAP GENERATION is selected and samples are defined the locations for each sample replicate must be specified LOCATION
152. r 1 Introduction BioTek Instruments Inc Chapter 2 Instrument Description This chapter includes principles of operation and descriptions of important components of the multi detection reader PRINCIPLES OF Opera Nina 10 External COMPONENTS ciar 10 10 Chapter 2 Instrument Description Principles of Operation The uQuant M is a single channel microplate spectrophotometer The instrument has a long life xenon flash light source a holographic grating based monochromator and UV grade fiber optics and lenses This combination allows for absorbance measurements in a spectral range from 200 nm to 999 nm in 1 nm increments The instrument bandpass is 2 4 nm Assay definitions along with the data they produce are managed by an onboard processor via a 2 line x 24 character LCD screen and membrane switch Data can be stored onboard printed and or uploaded to controlling software on a host PC The u Quant is designed to serve as a stand alone system or as part of a larger laboratory data network sending receiving and manipulating assay data as needed External Components e The power switch is located on the right side of the instrument The switch is labeled with I O indicating on and off respectively e The front panel features a 2 line x 24 character Liquid Crystal Display LCD and a 25 key keypad Through this interface you can create and modify absorbance assays read plates store and print results send re
153. racy linearity mechanical alignment repeatability and wavelength accuracy of the instrument e Liquid Testing page 117 quantifies the instrument using liquids which verifies operation in a way that glass test filters cannot D Set the uQuant to Normal Standard Read Mode when running all verification procedures Chapter 3 BioTek Instruments Inc Qualification Procedures 101 System Test IMPORTANT Do not turn on the instrument until the carrier shipping block has been removed The System Test is performed automatically whenever the instrument is turned on It can also be performed manually through the Quant Main Menu The System Test feature conducts a series of tests at each of six set wavelengths which confirm adequate light levels low electronic noise adequate photodiode sensitivity and overall system cleanliness The testing is designed to ensure that the Quant will give in specification performance for each set wavelength over the specified OD range The reader will chirp repeatedly if the power on System Test results do not meet the internally coded Failure Mode Effects Analysis FMEA criteria established by BioTek A system test should then be initiated to try to retrieve an error code from the reader To run the System Test from the instrument keypad 1 If running a System Test attach a printer to the instrument See Connecting a Printer to the pQuant in Chapter 4 for instructions 2 From
154. re BioTek Instruments Inc Appendix B Using 384 Well Plate Geometry When used in standalone mode the uQuant does not have access to data reduction options for 384 well plate geometry To capture and manipulate the raw data you must use Gen5 KC4 or KCjunior software This Appendix provides instructions for setting up and performing a 384 well plate read for data reduction in Gen5 KC4 or KCjunior Reading a 384 Well Plate in Gen5 Lo cceccecenseneueeseusevenseuereeneues 178 Reading a 384 Well Plate in KC4 cc ccececceeseeeeeeeeeeeeeeneeeeeeneags 182 Reading a 384 Well Plate in KCjunior 178 Appendix B Using 384 Well Plate Geometry Reading a 384 Well Plate in Gen5 1 Start Gen5 Log in if prompted Gen5 Secure users 2 When the Welcome to Gen5 screen appears select System Menu 3 At Gen5 s main screen select System Reader Configuration to open the Reader Configuration dialog Reader Configuration i X Reader Settings status Add view Todi s Close Help Figure 20 Reader Configuration dialog Actes Version 4 Click the Add button to open the Reader Settings dialog Reader settings x Reader Type TI Test Comm Lom Part am Setup Baud Fate seoo Cancel Help Figure 21 Reader Settings dialog 5 Use the drop down list to select uQuant and enter the appropriate Com Port gt The Baud Rate is set to the defau
155. reen will appear Press ENTER to cycle through the prompt screens SAMPLE UQuant Operator s Manual 96 Chapter 4 Operation gt If selected at read time PLATE ID prompts for microplate identification SAMPLE ID prompts for identification for each sample group SAMPLE COUNT prompts for the number of samples on the plate RAPID MODE prompts for the plate to be read in Fast Mode or Regular Mode RAPID MODE reads a 96 well plate in approximately 30 seconds If NO is selected the plate will be read in NORMAL MODE approximately 50 seconds Refer to Chapter 1 for specifications gt Pressing ENTER after each selection advances the display gt When selections are completed the display returns to the SELECT UTILITY OPTION menu BioTek Instruments Inc Chapter 5 Instrument Qualification This chapter discusses the tasks and procedures necessary for verifying and qualifying instrument performance on an ongoing basis Recommendations for Achieving Optimum Performance 98 Recommended Qualification Schedule een 99 Qualification Procedures essssen nemen emnes nnn 100 DV SEC BA TOS Deja biu des IM SD Nu ADD DUM a UE 101 CHECKSUM Tests 107 Absorbance Plate Testa ad 108 See NCS LEO CCE Em AO 121 EAU OSEA is io 123 BOU TOS Coria 127 98 Chapter 5 Instrument Qualification Recommendations for Achieving Optimum Performance Microplates should be perfec
156. ror Codes This appendix introduces the test protocols necessary for IQ OQ PQ Qualification This appendix also describes problems that you may experience with the uQuant during testing lists potential error codes and suggests possible solutions for these problems OVClVICW sidad iio 138 System est iDescriptulo Masia bici ias 138 Glossary of Temen nica jvc br 138 EMO COIG S actas 139 General Error S an T N E 140 Fatal EITO Soer o de a e DE DID PLE 163 138 Chapter 7 Troubleshooting and Error Codes Overview If an error code appears on the display or in Gen5 KC4 or KCjunior software consult the error code list to see if the error is easily correctable If you cannot resolve the problem run the instrument System Test and note the exact error code and serial number of the reader Then call BioTek s Technical Assistance Center refer to Chapter 1 for contact information System Test Description The System Test feature conducts a series of tests at each of six set wavelengths which confirm adequate light levels low electronic noise adequate photodiode sensitivity and overall system cleanliness The testing is designed to ensure that the Quant will give in specification performance for each set wavelength over the specified OD range The reader automatically runs an internal System Test each time it is powered on The reader will chirp repeatedly if the power on System Test results do not meet the intern
157. rror Codes 149 General Errors Continued Measurement channel light beam saturated too much light Air reading reached 65535 Either during a read or system test one of the filters from one to six has saturated the measurement channel The last number is the lambda table position number Probable Causes The monochromator mirror grating has a defect Analog PCB intermittently failed The order sorting filter wheel is jammed and not aligning the correct filter to line up with the light path or the through hole is allowing white light to pass Voltage to the lamp has increased because the lamp power supply has failed or the motor power supply PCB sent the incorrect voltage request to the lamp power supply Measurement channel gain out of range for the selected wavelength Fails if Air reading is gt 60000 with a gain of 1 While calibrating a filter or testing the reader before a read one of the filters saturated the measurement channel The last number is the lambda table position number Probable Causes The monochromator mirror grating is damaged The order sorting filter wheel is jammed and not aligning the correct filter to line up with the light path the filter is degraded and not passing enough light energy or the filter is blocking the light Analog PCB intermittently failed Missed flashes or erratic flash lamp Voltage to the lamp has increased because the lamp power supply has failed or the motor
158. rs AVELENGTH 40 5 WEL IL ALIBRATION VALUE 5 Enter the calibration values listed on the test plate s Standards Certificate After each entry press ENTER to advance to the next consecutive well location 6 When all values have been entered press the Main Menu key For the Peak Wavelength Test Note the test range specified in the Peak Wavelength Certificate then define a Spectral Scan assay Assay 00 to scan the C6 filter across the specified range in 1 nm increments Refer to Selecting an Assay in Chapter 4 for detailed instructions on setting up an assay BioTek Instruments Inc Qualification Procedures 111 To enter the test plate data into Gen5 select System Diagnostics Test Plates Add Modify Plates then click Add Click the Help button for guidance when setting the wavelengths and entering the OD and peak wavelength value s To enter the test plate into KC4 select System Diagnostics Define Universal Plates then click Add Click the Help button for guidance when setting the wavelengths and entering the OD and peak wavelength value s To enter the test plate data into KCjunior select Utilities Diagnostics Universal Plate Test then click New Data Sheet Click the Help button for guidance when setting the wavelengths and entering the OD and peak wavelength value s Running the Absorbance Plate Test D IMPORTANT Before running the Absorbance Plate Test ensure that the reader is not r
159. s shown that the device is accurate at these OD values then by definition it has to be linear Note that there may not be a Pass Fail indication for filter values that are beyond the specified accuracy range of the instrument If the reader fails the accuracy test review the following possible problems and solutions gt Check the neutral density filters on the test plate to ensure they are clean If necessary clean them with lens paper Important Do not remove the filters from the test plate and do not use alcohol or other cleaning agents gt Ensure that the filter calibration values entered via the keypad Gen5 KC47M or KCjunior are the same as those on the Test Plate Standards Certificate Ensure that the Test Plate is within its calibration certification period If these remedies are ineffective or if the Test Plate requires recalibration call your BioTek representative or BioTek Technical Support BioTek Instruments Inc Qualification Procedures 115 e Repeatability This test ensures that the instrument meets its repeatability specification by reading each Test Plate neutral density filter twice with the filter in the same location Note that there may not be a Pass Fail indication for filter values that are beyond the specified accuracy and thus repeatability range of the device If the reader fails the repeatability test review the following possible problems and solutions gt Check the ne
160. sults to an external computer and more See Chapter 4 Operation for more information e The microplate carrier supports the microplates and adapter plates described in Chapter 1 under Specifications A spring clip holds the plate securely in place The microplate carrier access door with its magnetic closure helps to ensure a light impermeable measurement chamber e When a plate read is initiated the plate carrier is drawn into the measurement chamber and then moves in the X and Y axes to align each microwell with the probe as specified in the assay When the read is complete the plate carrier is returned to its full out position e The back panel contains the power supply jack parallel printer port and serial port See Setting Up the Quant in Chapter 3 for more information BioTek Instruments Inc External Components 11 Front Door Microplate Carrier Power Switch Front Panel Serial Port Parallel Port Printer Port Power Supply Jack Figure 1 External components of the uQuant UQuant Operator s Manual 12 Chapter 2 Instrument Description BioTek Instruments Inc Chapter 3 Installation This chapter includes instructions for unpacking and setting up the UQuant and instructions for connecting printers and or serial devices Unpacking and Inspecting the uQuant een 14 setting WO THe Ola tana EET 20 operating EnvIFOTnent czotcsetdsansolsshelsentolcsabice
161. tart at the Main Menu and press UTIL gt TESTS gt SYSTEM See System Test and Checksum Test in Chapter 5 or Connecting a Printer to the pQuant on page 24 for more information HQuant Main Menu Following successful power up of the uQuant the Main Menu appears 01 22 PM 01 23 04 DEF INE REPORT U TI L e The Main Menu permits access to all onboard functions See Main Menu in Chapter 4 for more information including a diagram showing the flow of functionality e The Quant front panel contains four circular buttons referred to in this manual as SOFT KEYS One SOFT KEY is positioned directly below each selectable option in the display To select a menu option simply press its corresponding SOFT KEY See Quant Front Panel in Chapter 4 for additional instructions UQuant Operator s Manual 22 Chapter 3 Installation Adjusting the Wavelength Table The pQuant has six default wavelength settings installed at the factory Before operating the instrument configure the wavelength table with values that will confirm that the instrument is operating near its limits The recommended wavelengths are 200 405 490 550 630 and 999 nm You may choose values other than the upper or lower limits of the instrument if the Quant will never be used in these outer ranges Note that you may also choose wavelength values that coincide with six of the wavelengths 405 to 750 nm given in the BioTek Absorbance Test Plate PN 7260522 with
162. the RS 232 cable connection gt Baud Rate 1200 2400 or 9600 default must match the baud rate on the reader gt Data Bits 8 gt Parity None gt Stop Bits 2 gt EOT Character Keep the default number Although the baud rate may be changed to 1200 or 2400 BioTek recommends keeping the default rate of 9600 UQuant Operator s Manual 174 Appendix A Controlling the Quant with Gen5 KC4 or KCjunior Software 7 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 is displayed an incorrect COM port may have been selected Select a different port and then repeat this step Problems gt If the test passes at The communications test was successful dialog click OK to close the dialog then click OK again to save the settings and close the Reader Setup dialog gt If the test fails follow the directions provided by KCjunior then click Test Communications again See the Problems section below 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 in Table 7 and is not a Null cable If this is suspected add another Null and try again Getting Started with KCjunior The following instructions briefly describe how to read a plate using KCjunior Refer
163. the ASSay INaErie asus pd Doc eed a rS a s SAI 42 Define Method Map Formula and Curve ccccccccecseccesecceeeeseueeeeneags 43 Deling METHOD AAA 43 Br iginiers Ely ER 47 Den MING FORMULA dia 62 pelhng CURVE aa 74 Reading a Microplate il UR 83 Selecting an Assay to RUN ccccscceeecreescenseeeeessccneenteesseuuaateesteeunannes 84 P PINGING REDO Scerri a esu tan Odd A edd Gen tK Lc 88 Editing Standard OuUTICES o aii 89 PRPEING RESUS sad 91 A 92 Setting the Date and Time eene rn rr rr 92 Editing the Wavelength Table eee mnn 93 Specifying Data Output and Reporting Options eeeeeeeee 93 Selecting REAG OPUS aia 95 36 Chapter 4 Operation HQuant Front Panel Liquid READY 01 22PM 01 23 04 Crystal Display LCD READ DEFINE REPORT UTIL Menu options Soft keys 4 7 84 9 Previous S ial Alphanumeric 4o Je 6r function keys k S la 28 3c LOSI Press the Shift key and an alphanumeric key simultaneously to select a letter from A to H Figure 13 uQuant front panel with 2 line x 24 character LCD and keypad BioTek Instruments Inc uQuant Front Panel 37 The keypad has four SOFT KEYS one below each selectable menu option Press a SOFT KEY to make a selection For example from the Main Menu press the leftmost SOFT KEY to select READ the rightmost to select UTIL Exit the current screen and return to the Main Menu Pressing Main Menu while defining or
164. the CE mark Directive 89 336 EEC Electromagnetic Compatibility Emissions Class A EN 50081 1 1992 and IEC 61326 1 1997 EN 55022 1995 Class A Immunity EN 50082 1 1997 and IEC 61326 1 1997 EN 61000 4 2 1995 Electrostatic Discharge EN 61000 4 3 1996 Radiated EM Fields EN 61000 4 4 1995 Electrical Fast Transient Burst EN 61000 4 5 1995 Surge Immunity EN 61000 4 6 1996 Conducted Disturbances EN 61000 4 11 1994 Voltage Dips Short Interruptions and Variations Directive 73 23 EEC Low Voltage EN 61010 1 1993 Including Amendment 2 1995 Safety requirement for electrical equipment for measurement control and laboratory use Part 1 General requirements Directive 2002 96 EC Waste Electrical and Electronic Equipment Disposal Notice This instrument contains printed circuit boards and wiring with lead solder Dispose of the instrument according to Directive 2002 96 EC on waste electrical and electronic equipment WEEE Quant Operator s Manual XX 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 eq
165. the currently selected assay DEFINE METHOD MA P FORMULA CURVE Press the SOFT KEY beneath the displayed option to access the following functions e METHOD Specify the wavelength type wavelength s and plate geometry e MAP Specify mapping formation e FORMULA Access the formula entry screens e CURVE FIT Specify curve fit options Defining METHOD The definition of a method includes selecting e Single or dual wavelength e Wavelength s e Plate geometry D The options appear on the display in the order that they were programmed in the assay If the assay contains a closed variable i e an element of the assay definition that you cannot access or modify the entry screen is skipped UQuant Operator s Manual 44 Chapter 4 Operation Single or Dual Wavelength Note Only the Endpoint read method is supported here There is one spectral scanning onboard assay available see page 81 for more information WAVELENGTH SINGLE DUAL e Select SINGLE or DUAL wavelength e Press ENTER to continue The WAVELENGTH selection screen allows you to select SINGLE or DUAL wavelength for the assay If SINGLE wavelength is chosen the reader measures the optical density of each well at a single wavelength If DUAL wavelength is chosen each well is read twice each time at a different wavelength The microplate is not removed from the reading chamber between the two measurements The final reported optical density
166. tly 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 Although the uQuant M supports standard flat U bottom and V bottom microplates optimum performance is achieved with optically clear flat bottomed wells Nonuniformity 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 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 Dispensing solution into 384 well plates often traps air bubbles in the wells which may result in inaccurate readings A dual wavelength reading method usually eliminates these inaccuracies however for best results remove the air bubbles by degassing the plate in a vacuum chamber before reading The inclination of the meniscus can reduce reading accuracy in some solutions especially with small volumes Agitate the microplate before reading to help bring this problem within acceptable li
167. to clear the NUMBER OF BLANKS value from the display BioTek Instruments Inc Define Method Map Formula and Curve 55 Blank Location The BLANK LOCATION screen allows you to define where the blank well or wells occur on the microplate This screen only appears if manual mapping was selected EN TER T H E LOCATION BLANK 1 e Use the numeric and alpha keys to enter a Blank Location based upon the plate geometry e Use the arrow keys to move the cursor to the next or previous editable field The cursor is positioned beneath the first editable field e Press ENTER to continue Number of Standards This option allows you to enter the number of standard groups that will be used in the assay Any previously defined value will be displayed on the screen lt Note If the number of standards is altered the number of replicates for the standard automatically reverts to 1 EN TER NUMBER OF STANDARD S e Use the numeric keys to enter the NUMBER OF STANDARDS The valid range depends on the selected curve fit method The maximum number of standards is 12 The minimumi is 4 for 4 P fit cubic cubic spline and logit log 3 for quadratic and 2 for linear and point to point e Press CLEAR to clear the value on the display e Press ENTER to continue UQuant Operator s Manual 56 Chapter 4 Operation Number of Standard Replicates This option allows you to enter the number of replicates per standard gro
168. to create protocols assign well identifiers read plates print results perform data reduction and more To perform a Quick Read 1 At the Welcome to Gen screen select System Menu then at the main screen select File New Experiment Alternative select Read a Plate at the Welcome screen then proceed to step 4 on the following page Click Default Protocol then click OK Gen5 will open the Experiment workspace which includes the Protocol menu tree and Plate screen BioTek Instruments Inc Controlling the Reader with Gen5 169 3 Select Plate Read or click the Read Plate icon The Procedure dialog will open gt Gen5 and Gen Secure If more than one reader was added in Gen the Instrument Selection dialog will appear instead of the Procedure dialog Select uQuant then click OK The Procedure dialog will then appear Select a Plate Type Click Read to open the Read Step dialog Select a Read Type Define the wavelength s at which the plate will be read et x 2 Define other reading parameters as desired Click the Help button for assistance 9 When complete click OK to return to the Procedure dialog Click Validate if you would like Gen5 to verify the defined parameters If all parameters are valid you will receive confirmation If any parameters are invalid Gen5 will provide information for correcting the problem Refer also to the Troubleshooting section
169. to turn to the open hole UQuant Operator s Manual 142 Chapter 7 Troubleshooting and Error Codes General Errors Continued Saturation transition failed in the Y axis movement light beam never found During the Y axis movement the light beam saturation transition max light to no light was never found Probable Causes e Dirty rail and nylon bushings or bearings causing the carrier to jam e Autocal jig is not in the carrier e Order sorting filter wheel is jammed and unable to turn to the open hole Saturation transition failed in the monochromator motor movement light beam never found During instrument initialization the monochromator is homed by rotating the monochromator mirror until the white light full light is detected This requires a fully functional flash lamp detection system Probable Causes Defective flash lamp and or flash lamp power supply inconsistent flashes Defective analog PCB Defective motor power PCB Defective monochromator low probability Carrier X axis movement failed positional verify Motor X axis failed to get to the same position when moved a known number of steps from the home position and back Probable Causes e The optical trigger flag has moved or is loose e Dirty rail and nylon bushings or bearings e Carrier support pin is out of adjustment BioTek Instruments Inc Error Codes 143 General Errors Continued Carrier Y axis mov
170. to use BioTek s Gen5 KC4 or KCjunior for microplate data analysis and control software See Appendix B Using 384 Well Geometry for more information UQuant Operator s Manual 46 Chapter 4 Operation Press MORE to cycle through the available options 6 6 well 2x 3 60T 60 well Terasaki 6 x 10 12 12 well 3 x 4 72T 72 well Terasaki 6 x 12 24 24 well 4 x 6 96T 96 well Terasaki 8 x 12 48 48 well 6 x 8 96H 96 well Hellma Quartz 8 x 12 96 96 well 8 x 12 96M 96 well Metric 8 x 12 9 mm well 384 384 well 16 x 24 spacing BioTek Instruments Inc Define Method Map Formula and Curve 47 Defining MAP The MAP DEFINITION screen allows you to edit or specify the following options in the assay Automatic or manual map generation Mapping direction Replication direction Blank Map selection Blanking constant Number of blanks Location of blanks Number of standards Number of standard replicates Averaging of standards Concentration and location of standards Number of controls Control Type definition Number of control replicates Control location Number of samples Number of sample replicates Sample location IMPORTANT When 384 well plate type is selected access to map formula or curve options is denied The 384 well reads are preset for the map to have 384 samples This does not include any blanking controls or standards UQuant Operator s Manual 48 Ch
171. tor instrument light levels one can plot this value over time e g monthly for a given set of wavelengths e At low light levels Resets 1 if the Gain is over 20 you may want to contact BioTek Technical Support to schedule preemptive service on the instrument The instrument will continue to operate at these signal levels and pass the internal system test but preemptive service is advised Air Dark Value Just before each flash the Dark reading is taken representing the background signal in the instrument This dark is subtracted from a reading taken on Air with no plate in the light path The difference between the Air and Dark readings represents a true level of light available for measurements If this level drops below a certain threshold on any channel the Self Test fails UQuant Operator s Manual 106 Chapter 5 Instrument Qualification Noise Test The noise test ensures that there is minimal variation in signal between Dark readings with no light present The noise test values are given in the lower section of the System Test report The Noise test could fail if there is e Anambient light leak or electrical noise penetrating the measurement chamber The bottom and top shrouds provide light and electrical shielding Make sure that the plate chamber door is fully closed and the top and bottom shrouds are properly installed and fastened without any gaps e Internal electronic noise caused by a f
172. ts repeatability specification by reading each Test Plate neutral density filter twice with the filter in the same location e Wavelength Accuracy To check the wavelength accuracy use BioTek s 7 Filter Absorbance Test Plate PN 7260522 This test plate provides a multiband test filter in location C6 for the Peak Wavelength Test BioTek s 6 Filter Absorbance Test Plate PN 9000547 does not have a filter in the C6 location This test plate can be used on any BioTek reader however if this plate is used the Peak Wavelength Test will not be performed BioTek Instruments Inc Qualification Procedures 109 Test Plate Certificates To run the Absorbance Plate Test on the uQuant you will need BioTek s 7 Filter Absorbance Test Plate Part Number 7260522 with its accompanying certificates e The Standards Certificate contains standard OD values for the filters at several different wavelengths See the sample Standards Certificate in Figure 17 below e The Peak Wavelength Certificate contains one or more Peak Wavelength values for the glass filter in position C6 on the plate Each value has a valid test range associated with it For example a Peak Wavelength value may be 586 nm with a test range of 580 to 590 nm or tolerance values of 6 4 See the sample Peak Wavelength Certificate in Figure 19 on page 116 This test plate can be used for testing the reproducibility linearity and alignment of your BioTek aut
173. ts saved for that wavelength during normal or rapid mode The reference channel failed lt 100 or gt 20000 during filter calibration or spectral scan with the flash on The reference channel failed lt 100 during filter calibration or spectral scan with the flash off Probable Causes e If failed lt 100 Absorbance analog PCB or reference channel analog PCB is defective Shielding of the cable between reference channel and analog PCB is defective or disconnected Reference channel photodetector is defective If failed gt 20000 Reference channel photodetector is defective A faulty analog PCB or faulty internal grounding may cause internal electronic noise There may be an ambient light leak Ensure that the plate carrier door is properly closed Electrical noise may be penetrating the measurement chamber The bottom and top shrouds are part of the electrical shielding Check to see if the shrouds are installed and are properly fastened Order sorting filter wheel is jammed and not aligning the filter wheel to block the light path or the filter is degraded and not passing enough light energy UQuant Operator s Manual 154 Chapter 7 Troubleshooting and Error Codes General Errors Continued 0901 0906 The absorbance reference channel Dark current value failed See Dark on the system test See criteria below The last number is the lambda table position number This error indicates one of the fo
174. uipment 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 prescrites 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 North America e Canadian Standards Association CAN CSA C22 2 No 1010 1 92 Safety Requirements for Electrical Equipment for Measurement Control and Laboratory Use Part 1 General Requirements International e EN 61010 1 1993 and Amendment 2 1995 Safety Requirements for Electrical Equipment for Measurement Control and Laboratory Use Part 1 General Requirements B
175. umn as the first well of each sample even if the ACROSS direction has been specified for replicates 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 Manual mapping is recommended to set up the appropriate map by placing the standards controls and samples in only the odd columns 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 Manual mapping is recommended to set up the appropriate map by placing the standards controls and samples in only the A C E and G rows UQuant Operator s Manual 54 Chapter 4 Operation Constant Blank Value This entry screen only appears when a CONSTANT BLANK map is selected Enter the value to be subtracted from each well on the plate EN TER BLAN KING CONS TANT e Use the numeric keys to enter the value The range is 0 000 to 3 000 The cursor is positioned at the first editable field and advances automatically e Press CLEAR to clear the value on the display Press ENTER to continue Number of Blanks The NUMBER OF BLANKS field allows you to enter the number of blank wells on the plate This entry screen is only displayed when Full Column or Row Blank maps are selected Any previously defined value is displayed EN TER NUMBER OF BLANKS e Use the numeric keys to enter the number of blanks The range is 0 to 48 e Use the CLEAR key
176. unning in Rapid Mode To check this select UTIL gt READ and then cycle through the prompts until READ IN RAPID MODE is displayed Choose NO for an accurate result To run the Absorbance Plate Test from the instrument keypad 1 Startat the Main Menu and select UTIL gt TESTS CALPLATE The SELECT TEST menu will appear SELECT TEST SYSTEM C HK SUM CALPLA T E 2 Select the appropriate wavelength at the CALIBRATION LAMBDA screen and press ENTER to save the value and continue CALIBRATION LAMBODA 4 0 5 45 0 49 0 3 When prompted place the Absorbance Test Plate on the plate carrier and press the READ key to begin the test The Absorbance Test Plate report Calibration Plate Analysis will be sent to a printer when the test is run see Figure 18 for a sample report UQuant Operator s Manual 112 Chapter 5 Instrument Qualification To run the test in Gen5 select System Diagnostics Test Plates Run Select the appropriate reader if the Instrument Selection dialog appears Select the appropriate test plate if the Select Test Plate dialog appears Enter the reader Serial Number and User name required information then click Start Test To run the test in KC4 select System Diagnostics Run Universal Plate Test Select the appropriate Universal plate enter the reader Serial Number then click Run Test To run the test in KCjunior select Utilities Diagnostics Universal Plate Test Sele
177. up in the assay Any predefined value appears on the display EN TER NUMBER OF STANDARD REPLICATES e Use the numeric keys to enter the NUMBER OF STANDARD REPLICATES The range is 1 to 8 replicates The software will verify that the number of replicates multiplied by the number of standards does not exceed the number of wells on the plate e Press CLEAR to clear the value on the display e Press ENTER to continue Average Standards The AVERAGE STANDARDS option allows you to select whether or not to average the replicates of each standard group This average is used to calculate the standard curve instead of using the individual replicates of each standard If the number of standard replicates is 1 this option is not available AVERAGE STAN DARD S Y E Y E S N O 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 e Press ENTER to continue BioTek Instruments Inc Define Method Map Formula and Curve 57 Standard Concentrations The Standard Concentration field allows you to enter the predicted or expected concentration value for each standard group If manual mapping was selected the replicate locations must also be defined LOCATION RE P eC SO e Use the numeric and alpha keys and the decimal point key to enter stand
178. utral density filters on the test plate to ensure that there is no debris that may have shifted between readings and caused changes Check the microplate carrier to ensure that it is clear of debris gt If these remedies are ineffective call your BioTek representative or BioTek Technical Support e Wavelength Accuracy The 7260522 Test Plate has a glass filter in position C6 that is used to check wavelength accuracy of the monochromator The Test Plate Peak Wavelength Certificate lists the certified wavelength s at which the maximum absorbance peak occurs Gen5 KC4 and KCjunior each have an option that can be enabled to scan the region of interest during the Absorbance Plate Test and confirm the accuracy of the glass reading Note You can use the instrument keypad to run a spectral scan assay Assay 00 The resulting printout will indicate the wavelength at which the maximum absorbance peak occurs thus confirming the wavelength accuracy The C6 filter is scanned across a specified wavelength range in 1 nm increments The wavelength of the maximum absorbance is compared with the peak wavelength values entered via the keypad Gen5 KC4 or KCjunior which comes from the Peak Wavelength Certificate supplied with the test plate The accuracy of the wavelength should be 3 nm 2 nm instrument 1 nm filter allowance For example if the test range is 580 to 590 nm the Certificate value is 587 nm and the reader reports a peak
179. ween the reference channel and the absorbance analog PCB may be floating or not connected There may be an ambient light leak Ensure that the plate carrier door is properly closed Analog PCB failure the photodetector is noisy A faulty analog PCB or faulty internal grounding may cause internal electronic noise BioTek Instruments Inc Error Codes 151 General Errors Continued Absorbance reference channel failed offset range See noise Max lt 20000 and noise Min gt 10 on the system test During self test the background electronic signal detected is outside of acceptable limits at maximum gain when blocking the light Probable Causes e If noise Max is gt 20000 gt The photodetector is too noisy and defective Absorbance channel analog PCB is defective A faulty analog PCB or faulty internal grounding may cause internal electronic noise There may be an ambient light leak Ensure that the plate carrier door is properly closed Electrical noise may be penetrating the measurement chamber The bottom and top shrouds are part of the electrical shielding Ensure that the shrouds are installed and properly fastened e If noise Min is lt 10 gt The photodetector is not connected or is defective producing a noise reading of zero Analog PCB is defective UQuant Operator s Manual 152 Chapter 7 Troubleshooting and Error Codes General Errors Continued Measurement

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