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ELx800™ - Probiotek

1. 109 Sample Absorbance Test Plate Data Sheet 112 Sample Test Plate Analysis Report cccccsscceeseeeesseeeseenaees 112 Lamp Assem DIY aussessaukxashyus ades saa xa kat ER MM Y GEAR Mi RO E T EA 133 KCjunior Main Menu Setup Reader 1 Dialog 156 Reader Setup 1 Dialog ssdetssxeixaXait EXE dMebK E dstunzRx UM ERdds RN VULER 156 Read Plate Dialog 1 isssesziasuritxrnetaeciadaxatssaweriasbriukena ead Ee dabas 157 Protocol Definition General Information Dialog 157 Protocol Definition Read Method Dialog sess 158 KC4 Main Menu System Readers eren 159 Reader Selection Dialog 5 2 ore rn kr nad i nnn nna as 159 Data New Plate DIAlGE seus cian ea adag a ias pad Rat adea caa aoa TA UIS RPETiS 160 New Data File Dialog sierra re o e han n Redon da 160 Reading Parameters Dialog ccccccssceeeseesseeesseeessseeeseenaees 161 ELx800 Operator s Manual x Preface List of Figures Continued Figure 31 Figure 32 Figure 33 Figure 34 Figure 35 Figure 36 Figure 37 Samples With Calls on Matrix Report cccccsseeeeee esse tease eeaeeees 164 Curve Fit REDOT iie t oar een termine AE Aa AEAN 165 Samples With Calls on Column Report ccccece cece cess eee ee eee eee 166 Column Report Without Samples eene 167 Panel REPOM zie cseuemnaue is aida ERR RAKCP
2. 139 OOVEEVIGUI Ge oixaetias ose x bud S cantare Kiosk natale cma Cea EENE Xica ERIT RC aaepe n CE w sicat ESAE 140 System Test DESCHPUON Ius eee da ari xol at aO oe OA Oc ER RR e 140 Glossary Of Ferm isoiicadiovisd assa Ea ERR AER TERIAR Dua A ERRER A RARE A SPREPGPKR ERE 140 Erro ore 141 G n ral Errol S srr insin o i aute dua M PD E A E E PIDE ER Ms Uh dde 142 Fatal Errors enean a TE E EE E EE E acd 148 Appendix A Controlling the Reader With KCjunior or KC4 149 p al PT 150 Controlling the Reader With KCjunior seeesssseeseeeeee nnne nnne nnn 151 PFOBIGITIS 2er oed Du RH e En RC D aoi e da i DRE Ed EM DD b E ID HR v ERA d 152 Getting Started With KCJUNIOr ossa dad esa Usb E a oU at EDU ER S b Rae 152 Controlling the Reader With KGCA M iiu odise ne en kam anas u anna aka aano arx n Ras 153 PFODIGITIB irieoiecaex dites vdsaskakont ni ostad a IER XEUEP EE SR viu d AT ER ai dA EIS I RiD E 154 Getting Started With KC4 uuu oeosasese vis dani d bd dub qax dpa ve ger vba d sax T ooPaid are E rip 154 Appendix B Using 384 Well Geometry ees 155 LASNIETIITO Nm 156 8 PM pm 159 Appendix C Report Fornmiat iiiissascsssass miss E PREND DIMINUER ENNIUS 163 Appendix D Comparison of the ELx800 and the ELSOO 171 ELx800 Operator s Manual viii Preface Appendix E Instru
3. cesses nnne nnne nnn nn ELx800 Main Men 2 erre etel nana i xod ax haa n Configuring Global Default Options cc cece eeeeeeeeeeeeeeaees Connecting a Printer to the ELX800 cesser Setting Up the Serial Port for Communications With Other Devices Attaching the Cable agris er sau qux Xe pui axe Cosas svkntetuusaa E exe E EUM RA Setting Communication Parameters esssesseeeen nenne Installing Additional Filters coe s see uon toa eeu Ia a tue reru en aevo ze ES Repackaging and Shipping the ELX800 cccecceeeeeeeeee eee eeeee ees 24 24 14 Chapter 2 Installation Unpacking and Inspecting the ELx800 Important Save all packaging materials If the ELx800 is shipped to the factory for repair or replacement it must be carefully repackaged according to the instructions on pages 30 through 31 using the original packing materials PN 7332062 The individual packaging materials contained in PN 7332062 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 Bio Tek The ELx800 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
4. m Liquid Test 3 i id ve s GT RR aci co X GA E RA Va Na Gad T 104 Chapter 4 Performance Verification and IQ OQ PQ Procedures Recommendations for Achieving Optimum Performance Microplates should be perfectly clean and free of dust or bottom scratches Use new microplates from sealed packages Do not allow dust to settle on the surface of the solution use microplate covers when not reading the plate Filter solutions to remove particulates that could cause erroneous readings Although the ELx800 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 ina 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
5. 1 Calculate the mean and standard deviation for the five readings taken above at each concentration Only one data set needs to be analyzed for each concentration The well that shows the most variation for each concentration is selected for data reduction For each mean below 2 000 OD calculate the allowed deviation using the repeatability specification for a 96 well format of 0 5 0 005 OD from 0 000 to 2 000 OD 405 nm 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 0 5 1 951 0 005 0 0098 which when added to the 0 005 0 0098 0 005 0 0148 OD which is the allowable deviation Since the standard deviation is less than this value the reader meets the test criteria Repeatability Specification For comparison in this test the following repeatability spec is applied using Normal mode 96 well microplate 0 5 0 005 OD from 0 000 to 2 000 OD 9405 nm ELx800 Operator s Manual 124 Chapter 4 Performance Verification and IQ OQ PQ Procedures 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 Calcula
6. 49 Define Method Map Formula and Curve The DEFINE screen allows you to edit the Method Map Formula or Curve Fit parameters for the currently selected assay FORMULA CURVE Press the SOFT KEY beneath the displayed option to access the following functions e METHOD Specify the wavelength type wavelength filter s and plate geometry e MAP Specify mapping information e FORMULA Access the formula entry screens e CURVE FIT Specify curve fit options Defining METHOD Defining a method includes selecting e Single or dual wavelength e Filter s e Plate geometry 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 ELx800 Operator s Manual 50 Chapter 3 Operation Single or Dual Wavelength WAVELENGTH DUAL S INGLE DUAL 1 Select SINGLE or DUAL wavelength 2 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 with a single filter If DUAL wavelength is chosen each well is read twice each time with a different filter The microplate is not removed from the reading chamber between the two measurements The final reported optical density is the difference
7. px 8 4 818 2 8 9 9 23 stor sTD2 sts srD4 sts PC NC SMP SMP stor Stb2 sTb3 srD4 sts Pe NC SMP sme Bio Tek Instruments Inc Define Method Map Formula and Curve 57 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 START MAPPING A T WELL LOCATION AOl 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 ELx800 Operator s Manual 58 Chapter 3 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 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 Bio Te
8. 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 HBS G 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 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 Bio Tek Instruments Inc Printing Reports 95 e Select YES to exclude the replicate from curve fit calculations e Select NO to retain the replicate e Press ENTER to adv
9. To run the System Test using KCjunior open the Utilities menu and select Diagnostics Reader System Test To run the System Test using KC4 open the System menu and select Diagnostics Run Optics Test The System Test report lists results in a Pass Fail format Figures 16 and 17 and can be viewed saved and printed Print the report to document periodic testing or troubleshoot If the reader fails the System Test it will not perform a read until the error condition is cleared See Chapter 6 Troubleshooting and Error Codes for a list of possible error codes that may be displayed if the System Test indicates a failure ELx800 Operator s Manual 108 Chapter 4 Performance Verification and IQ OQ PQ Procedures Note This report may vary slightly if KC4 or KCjunior are used Operator ID Notes 11 00 AM 06 02 03 Lambda 200 Gain 3 28 Channel Ref 1 Air 20506 35689 Dark 9911 10002 Delta 10595 25687 Lambda 405 Gain 2 53 Channel Ref 1 Air 20072 35592 Dark 9914 9960 Delta 10158 25632 Lambda 490 Gain 1 72 Channel Ref 1 Air 20280 35642 Dark 9919 9949 Delta 10361 25693 Lambda 550 Gain 2 51 Channel Ref 1 Air 20248 35530 Dark 9914 9985 Delta 10334 25545 Lambda 630 Gain 3 94 Channel Ref 1 Air 20230 35302 Dark 9907 10018 Delta 10323 25284 Lambda 999 Gain 5 33 Channel Ref 1 Air 19977 34186 Dark 9900 10050 Del
10. From this menu press FILTER 4 The wavelength for Filter 1 will be displayed To advance to Filter 2 press ENTER 5 To change the filter 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 filter on the filter table Bio Tek Instruments Inc Installing Additional Filters 29 ENTER FILTER 1 WAVELENGTH When the last filter has been entered the software exits the filter routine and displays the following screen EDIT SETUP INFORMATION DATE T IME FILTER MORE 6 Press the Main Menu key to return to the main menu ELx800 Operator s Manual 30 Chapter 2 Installation Repackaging and Shipping the ELx800 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 IMPORTANT Failure to properly repackage the If you need to ship the ELx800 Reader to Bio Tek for service or repair 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 Bio Tek for replacement packing see Technical Assistance in Chapter 1 for contact information Warning If the
11. This screen only appears if manual mapping was selected LOCATION 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 Note If the number of standards is altered the number of replicates for the standard automatically reverts to 1 ENTER NUMBER O F STANDARDS 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 minimum 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 ELx800 Operator s Manual 62 Chapter 3 Operation Number of Standard Replicates This option allows you to enter the number of replicates per standard group in the assay Any predefined value appears on the display ENTER NUMBER O F 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 multipl
12. 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 ELx800 Operator s Manual 118 Chapter 4 Performance Verification and IQ OQ PQ Procedures Solution B Required Materials e Bio Tek QC Check Solution No 1 PN 7120779 25 ml or 7120782 125 ml e Deionized water e 5 ml Class A volumetric pipette e 100 ml volumetric flask Preparation of Stock Solution 1 Pipette a 5 ml aliquot of Bio Tek QC Check Solution No 1 into a 100 ml volumetric flask 2 Make up to 100 ml with DI water cap and shake well This should create a solution with an absorbance of about 2 000 when using 200 ul in a flat bottom microwell The OD value result will be proportional to the volume in the well and the amount of QC Check Solution No 1 used You can use a larger or smaller well volume or add more Check Solution or water to adjust the stock solution Note that too small a well volume may result in increased pipetting related errors Bio Tek Instruments Inc Liquid Testing 119 Liquid Test 1 A 96 well flat bottom microplate is required for this test Corning Costar 3590 is recomm
13. 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 1 through 4 on the following pages when removing the instrument and its accessories from the shipping container See Repackaging and Shipping the ELx800 at the end of this chapter for complete shipping instructions Packing Materials PN Outer shipping container 7332036 Inner shipping container accessories 7330002 Left foam end caps 7332060 Right foam end caps 7332061 2 Mil poly bag ELx800 98085 8 V x 11 bubble bag power supply 91083 Required Tools PN Slotted and Phillips head screwdrivers 98145 Bio Tek Instruments Inc Unpacking and Inspecting the ELx800 15 Unpacking the Instrument and Its Accessories 1 Carefully open the top of the box and remove the power supply shelf Figure 1 2 Remove the top foam end caps manual and Declaration of Conformity Figure 1 Removing the power supply and shelf ELx800 Operator s Manual 16 Chapter 2 Installation 3 Lift the reader out of the box and remove the foam end caps Figure 2 Place the reader on a level surface and rem
14. gt 150 ul of the 75 Test Solution into all wells of columns 3 and 4 gt 150 ul of the 50 Test Solution into all wells of column 5 and 6 Read the microplate five times using Normal Mode single wavelength at 340 nm no blanking or blank on air Print the five sets of raw data or export it to an Excel spreadsheet using KCjunior or KCATM 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 ELx800 Operator s Manual 128 Chapter 4 Performance Verification and IQ OQ PQ Procedures Repeatability Test 1 Foreach well calculate the mean and standard deviation of the five readings 2 For each mean calculated in step 1 calculate the allowed deviation using the repeatability specification for a 96 well plate in the Normal Read Mode which is 1 5 0 005 OD from 0 000 to 2 000 OD 340 nm mean 0 015 0 005 3 Foreach 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 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 5 0 8004 0 015 equals 0 012 and when added to the 0 005 0 012 0 005 equals 0 017 which is the allowed deviation for well A1 Since the standard devia
15. 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 Bio Tek Instruments Inc Define Method Map Formula and Curve 71 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 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 gt Tocycle 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 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
16. 0 004 SMP0009 ASSAY 01 F02 0 006 0 006 ASSAY 02 B06 0 007 0 007 SMP0010 ASSAY 01 G02 006 006 ASSAY 02 C06 0 010 0 010 SMPOO11 ASSN 01 H02 005 005 ASSAY 02 D06 0 004 0 004 SWP0012 ASSAY 01 A03 00 00 ASSAY 02 E06 0 003 0 003 SWP0013 ASSAY 01 B03 0 00 0 002 ASSAY 02 FOG 001 001 SMP0014 ASSAY 01 03 005 005 ASSAY 0 606 0 001 0 001 SMP0015 ASSAY 01 D03 002 002 Page 1 Figure 35 Panel Report Bio Tek 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 36 Assay Detail Report Sheet 1 of 2 169 ELx800 Operator s Manual 170 Appendix C Report Format Assay List Version 3 ASSAY OI ASSAY 02 ASSAY 03 ASSAY 04 ASSAY 05 ASSAY 06 ASSAY 07 ASSAY 08 ASSAY 09 ASSAY 10 ASSAY 11 ASSAY 12 ASSAY 13 ASSAY 14 ASSAY 15 ASSAY 16 ASSAY 17 ASSAY 18 ASSAY 19 ASSAY 20 ASSAY 21 ASSAY 22 ASSAY 23 ASSAY 24 ASSAY 25 ASSAY ASSAY 27 ASSAY 28 ASSAY 29 ASSAY 30 ASSAY 31 ASSAY 32 33 ASSAY 33 34 ASSAY 34 35 ASSAY 35 36 ASSAY 37 ASSAY 37 38 ASSAY 38 39 ASSAY 39 40 ASSAY 40 41 ASSAY 41 42 ASSAY 42 43 ASSAY 43 44 ASSAY 44 45 ASSAY 45 46 ASSAY 46 47 ASSAY 47 48 ASSAY 48 49 ASSAY 49 50 ASSAY 50 51 ASSAY 51 52 ASSAY 52 53 ASSAY 53 54 ASSAY 54
17. 000 to 2 000 OD 405 nm Linearity 2 5 from 0 000 to 2 000 OD 405 nm Repeatability STD 2 076 0 010 OD from 0 000 to 2 000 OD 405 nm ELx800 Operator s Manual 10 Chapter 1 Introduction Technical Support Bio Tek s ELx800 Absorbance Microplate Reader is backed by a superior support staff If the reader ever fails to work perfectly please contact Bio Tek s Technical Assistance Center TAC Whichever method of contact you choose please be prepared to provide the following information Product name and serial number The software configuration information To locate this information press UTIL TESTS gt CHKSUM from the reader s Main Menu The specific steps that reproduce your problem Any error codes displayed on the screen descriptions of error codes are available in Chapter 7 A daytime phone number Your name and company information A fax number and or e mail address if available If you need to return the reader to Bio Tek for service contact Bio Tek for a Return Materials Authorization number RMA repackage the reader properly see Chapter 3 Installation and ship the instrument to Bio Tek at the shipping address listed on the following page Phone Support You can telephone the Technical Assistance Center between 8 30 AM and 5 30 PM Eastern Standard Time EST Monday through Friday excluding holidays Bio Tek Instruments Main Number 802 655 4040 Technical Assista
18. 1 0 0 010 OD from 0 000 to 2 000 OD 405 nm Linearity 1 0 from 0 000 to 2 000 OD at 405 nm 3 0 from 2 000 OD to 3 000 OD 450 nm Repeatability STD 0 5 0 005 OD from 0 000 to 2 000 OD 405 nm e The following specifications apply to 96 well flat or round bottom plates single wavelength measurements with a 30 second read rapid read mode Absorbance Measurement Range 0 000 to 3 000 OD Accuracy 2 0 0 020 OD from 0 000 to 2 000 OD 405 nm Linearity 2 0 from 0 000 to 2 000 OD 405 nm Repeatability STD 1 0 0 010 OD from 0 000 to 2 000 OD 405 nm e Light Source Tungsten gas filled bulb e Dimensions 42 cm x 38 cm x 18 cm 16 5 deep x 15 wide x 7 tall e Weight 8 kg 18 5 Ib Maximum e Environment Operating temperature 18 to 40 C e Humidity 10 to 85 noncondensing e Power Supply Input 100 to 240 V 10 0 50 to 60 Hz Output 24 VDC 2 1 A Bio Tek Instruments Inc Specifications 7 Ultraviolet UV Model The following specifications apply to 96 well flat or round bottom plates Wavelength Range 340 to 750 nm Filters 10 nm half bandwidth interference filters User accessible filter wheel Up to 5 filters may be installed on the instrument at one time Filters supplied 340 nm 405 nm 450 nm 490 nm and 630 nm Optical specifications for the 400 to 750 nm range 50 second read in normal read mode Absorbance Measurement Range 0 000 to 3 000 OD Accuracy
19. 131 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 3 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 Usea clean dry cloth to dry all wet surfaces ELx800 Operator s Manual 132 Chapter 5 Maintenance and Decontamination Replacing the Bulb The lamp is rated for an average life of 600 hours The intensity of the lamp will slowly drop over time until the run time self check detects a low signal level and the instrument displays an error code The bulb should be replaced and aligned at this time use replacement bulb PN 7330513 For UV and NB readers use replacement bulb PN 7330516 Figure 20 shows a detailed view of the lamp assembly reader has been shut down Allow the lamp to cool down for Warning The lamp will be hot immediately after the at least 10 minutes before attempting to replace the bulb To replace the bulb 1 2 Important Turn off and unplug the instrument Turn the instrument over on its back and remove the screws holding on the top cover see Figure 3 in Chapter 2 Turn the instrument over again and lift the top cover off Use a Phillips head screwdriver to re
20. 2 064 F02 SMPQ012 2 3 214 G02 SMP0013 3 663 H02 SMP0014 1 045 A03 SMP0015 0 090 Figure 33 Samples with calls on Column Report Bio Tek Instruments Inc 167 Bio Tek Instruments Assay Open Assay 04 Date 03 16 04 Lot l Time 01 05 34PM Operator Wavelength 405 Temp Plate ID COMMENTS Interpretation of Results GREYZONE 0 20 20 200 CUTOFF 1 1 006 SMP SHP lt CUTOFF4 CUTOFFSGREYZONE AND SMP gt CUTOFF CUTOFF amp GREYZONE EQUIV SMP SHP gt CUTOFF CUTOFFEGREYZONE NEG SMP SMP lt CUTOFF CUTOFFAGREYZONE POS PC PC a Well 1D Calc0D Call Pred Conc RSLT Std Dev AQ PC J 0 108 VAL BOL PC 0 119 VAL AVE j 9 114 End of Report Figure 34 Column Report without samples ELx800 Operator s Manual 168 Appendix C Report Format Bio Tek Instruments Date 07 31 96 Lot Time 02 24 01PM Operator Temp Plate ID COMMENTS Speciaen Assay Well BIkdOD Calcd Call RSLT Std Dev CVS SMP0001 ASSAY 01 F01 006 008 ASSAY 02 BOS 0 000 0 000 SMP0002 ASSAY 01 60 0 002 0 002 ASSAY 02 C05 0 007 0 007 SMP0003 ASSAY O1 H01 0 005 0 005 ASSAY 02 D05 0 004 0 004 SMP0004 ASSAY 01 A02 0 002 0 002 ASSAY 02 E05 0 012 0 012 SWPO005 ASSAY 01 B02 002 002 ASSAY 02 F05 0 010 0 010 SWP0006 ASSAY 01 C02 0 005 0 005 ASSAY 02 G05 001 001 SMPQ007 ASSAY 01 D02 00i 001 ASSAY 02 HOS 0 002 0 002 SwP0008 ASSAY 1 E02 0 009 0 009 ASSAY 02 A06 0 004
21. 655 7941 Outside the USA 802 655 4740 E mail customercare biotek com www biotek com Creating Solutions for the Future of Science ELxs800 Absorbance Microplate Reader Operator s Manual October 2004 2004 Part Number 7331000 Revision N Bio Tek Instruments Inc ii Preface Notices Bio Tek Instruments Inc Highland Park P O Box 998 Winooski Vermont 05404 0998 USA All Rights Reserved 2004 Bio Tek 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 Bio Tek Instruments Inc Trademarks Bio Tek is a registered trademark and ELx800 KC4 KCjunior and EL800 are trademarks of Bio Tek Instruments Inc Microsoft Windows and the Windows logo are registered trademarks of Microsoft Corporation in the United States and other countries 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 Bio Tek Instruments Inc Changes made to the information in this document will be incorporated in new editions of the publication No responsibility is assumed by Bio Tek for the use or reliability of software or equipment that is not supplied by Bio Tek or i
22. Figure 13 Figure 14 Figure 15 Figure 16 Figure 17 Sheet 2 of 2 Figure 18 Figure 19 Figure 20 Figure 21 Figure 22 Figure 23 Figure 24 Figure 25 Figure 26 Figure 27 Figure 28 Figure 29 Figure 30 List of Figures ix List of Figures Removing the Power Supply and Shelf cccscecseseeseeeeseeeeess 15 Removing the Foam End Caps sosiusxeas asd va Siva da E UR b ERR 16 Removing the Top Cover Mounting Screws eeeeeeeees 17 Removing the Shipping BIOCK 4 posuikta edu axe Ra audax Y RE ARCU E ra dads 18 Global Configuration Options Available Under UTIL 21 Connectors for Printer Parallel Computer Serial and Power Mr mE 23 Filter Wheel Assembly uuasir anui ERE Aa M ACRI d Yu ERR s A E RR ca 26 Securing The Carrier as vasa sau ex uE da RA DA a d RE RM a x RR ED na Rt Valla 31 Hooking the Shipping Straps Around the Shaft 32 Installing the Shipping BlOCK i5 2552x222 33442 325 9oRKa uo naxRw YS KFRER E bu read Ea 23 Reinstalling the Top Cover oloiieeeeki nana ea nona aka aar aaa kan ka nana 34 Reattaching the End CaS eseoee erkennen basa kebve s etkase rre Gan Fekrsakes 35 Placing the Unit in the BOX esu uud A aaa nra VE Lr ape dE s 36 hicyzsis pem 40 Options Available from the Main Menu een 44 Sample Output for the System Test Sheet 1 of 2 108 Sample Output for the System Test Autocal Analysis
23. Help Last Modified f 0 5 2004 2 45 08 PM Protocol Description Hn m Min Max Limits Minimum Value Maximum Value Raw Data Limits 0D 3 E IV Discard Out of Range Values Blank Limits OD 3 g IV Discard Out of Range Values m Precision 3 Scientific Notation Raw Data Blanks Scientific Notation Transforms Concentrations Displayed Decimal Places Figure 24 Protocol Definition General Information dialog ELx800 Operator s Manual 158 Appendix B Using 384 Well Geometry 6 Define the Read Method parameters gt Set the Plate Geometry to 8 x 12 Protocol Definition Incubation emperature Rea C Pani DES Head Mode fe Standard Rapid Sweep Figure 25 Protocol Definition Read Method dialog 7 Click OK to read the plate and display the results Bio Tek Instruments Inc KC4 159 KC4 Bio Tek s KC4 data reduction package can control the ELx800 in 6 12 24 48 96 and 384 well formats as well as in 60 72 and 96 well Terasaki plate formats 1 Start KC4 2 AtKC4 s main menu choose System Readers sKC4 NewPlate created from New Test Protocol prt Data Edit Protocol Display X gt 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 74 Figure 26 KC4
24. Select a Read Method Type of Endpoint Kinetic or Multiwavelength 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 Print the raw data by selecting Plate Print Results gt To analyze or manipulate results a protocol should be defined Refer to KCjunior s Help system or User s Guide for instructions Bio Tek Instruments Inc Controlling the Reader With KC4 153 Controlling the Reader With KC4 The ELx800 can be operated using a computer running Bio Tek s KC4 software Follow the steps below 1 DU wg Power off the computer and the reader Connect the appropriate serial cable PN 75053 between the two machines Power up both machines Install KC4 on the computer s hard drive Once installed start KC4 Select System Readers Scroll through the list of Available Readers and select the appropriate ELx800 reader model Click the Port button and subsequent Setup button to define the following communications parameters Port COM1 2 3 or 4 the serial port used for the RS 232 cable connection gt Baud Rate
25. WELL ALIBRATION VALUE 5 Enter the values listed on the Test Plate Data Sheet After each entry press ENTER to advance to the next consecutive well location Repeat for the remaining filters 6 When all values have been entered press the Main Menu key ELx800 Operator s Manual 114 Chapter 4 Performance Verification and IQ OQ PQ Procedures To enter the calibration values into KCjunior 1 First define the Absorbance Test Plate to be used for the test To do this open the Utilities menu and select Diagnostics Universal Plate Test 2 Click New Data Sheet Click the Help button for guidance To enter the calibration values into KC4 1 First define the Absorbance Test Plate to be used for the test To do this open the System menu and select Diagnostics Define Universal Test Plate Add Plate Click the Help button for guidance Running the Absorbance Plate Test IMPORTANT Before running the Absorbance Plate Test 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 To run the Absorbance Plate Test from the instrument keypad 1 Start at the Main Menu and select UTIL gt TESTS gt CALPLATE he SELECT TEST menu will appear SELECT T EST SY ST EM CHEK SUM CALPLA T E 2 Select the appropriate wavelength at the CALIBRATION FILTER screen and pr
26. advances automatically gt To change 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 ELx800 Operator s Manual 98 Chapter 3 Operation Viewing Editing the Filter Table After installing new filters ensure that the ELx800 s filter table the reader s software reference for filter locations matches the physical location of the filters in the filter wheel To edit the filter table 1 From the Main Menu select UTIL gt SETUP The EDIT SETUP INFORMATION menu will appear SETUP INFORMATION T IME FILTER MORE 2 Select FILTER to display the wavelength for Filter 1 NTER ILTERIHI1 WAVELENGTH 3 Enter the correct wavelength for each of the five positions using the numeric keys The range is from 400 to 750 nm 340 to 750 nm for the ELx800NB 4 Continue to scroll through the five available filter positions pressing ENTER between each 5 When all five wavelengths have been defined or skipped the software will automatically return to the EDIT SETUP INFORMATION menu Bio Tek Instruments Inc Using the Utility Options 99 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
27. 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 xxii Preface A A e A an vo E Caution refer to accompanying documents Attention voir documents d accompanement Achtung siehe Begleitpapiere Atenci n vease los documentos incluidos Attenzione consultare la doc annessa Warning risk of electric shock Attention risque de choc lectrique Gef hrliche elektrische Shlag 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 hot surface Attention surface c
28. be greater than the mean of the negative controls The formula is PC x NC x Bio Tek Instruments Inc Define Method Map Formula and Curve 75 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 70 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 69 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 Formulas Simple transformation formulas are typically applied to all wells on the plate For example 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 TVAR can be defined for use within a transformation formula This variable defines the scope of the transformation whether to apply the transformation to all of the wells on the plate OD o
29. 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 The currently selected wavelength appears on the top line of the display and the available options appear on the bottom MEAS Selection The MEAS selection screen allows you to select the filter s for the assay 1 Select the wavelength 2 Use the right arrow key to move the cursor to REF and then select the Reference Filter 3 Press ENTER to move to the next screen Bio Tek Instruments Inc Define Method Map Formula and Curve 51 Plate Type For 6 to 384 well standard plates the plate types and sizes included in the software onboard the ELx800 are based on the brands listed below For best measurement results use these brands when operating the ELx800 via its onboard software Plate Type Brand 6 well Costar 12 well Corning 24 well Corning 48 well Costar 96 well Costar 384 well Nunc square 96 T Terasaki 72T Terasaki 60T Terasaki Used only on ELx800NB version 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 Note The NB version of the reader includes choices for 384 well or 72 and 96 well Terasaki plate formats An adapte
30. bulb left or right to get a full circular image on the carrier sheet metal Tighten the lamp retention screws and verify that the lamp has not moved After completing the alignment turn the instrument off and on again to return the carrier to the home position Warning The alignment procedure requires you to observe the light path while the bulb is turned on To prevent possible vision impairment avoid looking directly at the bulb while it is on To align bulbs on UV and NB model readers 1 Use the lead wires to position the bulb to avoid damaging the glass covering the bulb with fingertips Keep the tip of the bulb slightly away from the optics arm 1 aperture 1 mm Rotate and or swing the bulb left or right to get a good circular image on the carrier s sheet metal Tighten the lamp retention screws and verify that the lamp has not moved After completing the alignment turn the instrument off and on again to return the carrier to the home position Bio Tek Instruments Inc Decontamination Procedure 135 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 performin
31. checksum After a few moments a second screen will display 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 ELx800 Operator s Manual 110 Chapter 4 Performance Verification and IQ OQ PQ Procedures Absorbance Plate Test Note Before running the Absorbance Plate test ensure that the Calibration Plate values are entered See Entering the Absorbance Plate Specifications in this chapter for more information The Absorbance Test Plate or Test Plate can be used to conduct several useful tests in a short time period Mechanical Alignment Accuracy and Linearity and Repeatability The data and results for these tests can be reviewed on the Absorbance Plate Test report To run an Absorbance Plate Test you will need a Bio Tek Absorbance Test Plate PN 7260522 with its accompanying data sheet D Mechanical Alignment The Test Plate has 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 ELx800 may be out of alignment If the reader fails the alignment test review the following possible problems and solutions Ensure that the Test
32. filter or a blank plug Install all filters with the arrow denoting the light direction pointing downward Filter wheel cover Thumbscrew Optics arm Filter wheel Figure 7 Filter wheel assembly Bio Tek Instruments Inc Installing Additional Filters 27 If you wish to install additional filters or change the filter locations 10 11 12 13 14 Turn off the unit and disconnect the power cord Tape down the carrier in the home full out position to prevent movement while the instrument is being handled Carefully turn the instrument upside down on a level surface Using a slotted screwdriver remove the four screws from the bottom of the instrument see Figure 3 Hold on to both the instrument base and the top case to prevent the detachment of the case from the body of the instrument Carefully turn the instrument right side up and remove the top cover With the cover off the instrument s mechanics and optics are exposed The filter wheel is housed within the small black box filter wheel cover directly under the end of the optics arm Figure 7 Remove the four thumbscrews around the perimeter of the cover The filter wheel sits on a pin and can be lifted off Remove the filters by turning the wheel upside down over a cloth The filters which are labeled with the wavelength and light direction arrow should easily slide out When replacing the filters in the wheel handle them from th
33. 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 1 To specify data output and reporting options start at the Main Menu and select UTIL gt OUTPUT The REPORT OUTPUT screen will appear EPORT OUTPUT BOTH RINT COMPUTER BOTH 2 Thecurrent 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 Note These options have no effect on data output if the instrument is being controlled by software such as KCjunior or KC4 running on a host PC 3 Press ENTER to continue The SELECT PRINTER screen will appear SELECT PRINTER EPS ON H P The ELx800 supports printers using either HP s PCL3 language such as the HP DeskJet series or Epson s LQ language For the latest list of compatible printers consult the Bio Tek Web site www biotek com or call Bio Tek Instruments Technical Assistance Center refer to Chapter 1 for contact information 4 Select EPSON or HP as appropriate ELx800 Operator s Manual 100 Chapter 3 Operation 5 Press ENTER to continue The REPORT TYPE screen will appear REPORT TYPE COLUMN 6 The currently selected report type is displayed in the top line Select COLUMN to print information in a list columnar fo
34. lt 2 500 Or with two separate formulas PC gt 1 000 and PC 2 500 Negative Control mean must have an OD of less than 0 100 the formula is NC x 0 100 ELx800 Operator s Manual 74 Chapter 3 Operation 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 NUMBER O F 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 considered 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 MAP before creating assay validation formulas Examples If an assay protocol states that for the assay to be valid gt The mean of the negative controls must be less than 0 100 The formula is NC x 0 100 gt The mean of the positive controls must
35. microplate prepared above five times using Normal Read Mode dual wavelength at 450 nm with 630 nm as the blank Note Do not discard the plate you will use it for the Alignment Test 2 Print the raw data or export it to an Excel spreadsheet using KCjunior or KC4 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 Linearity Test 3 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 Select Tools 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 Bio Tek Instruments Inc Liquid Testing 123 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
36. 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 Bio Tek Instruments Inc Printing Reports 93 Printing Reports Reports are automatically generated after a plate has been read see Specifying Data Output and Reporting Options on page 99 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 e Note See Appendix C for sample reports 22 PM 01 23 04 IN IE REPORT UTI L e Select RESULT to print an exact copy of results from the plate reading the 8 e 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 formulas and curve fit parameters e Select LIST to print a list of all assays name and number currently programmed in the ELx800 ELx800 Operator s Manual 94 Chapter 3 Operation Results Report
37. reader has been exposed to potentially hazardous material decontaminate it to minimize the risk to 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 See Chapter 5 Maintenance and Decontamination for the Decontamination procedure 2 Once the reader is clean follow the instructions on the next few pages to repackage the instrument Bio Tek Instruments Inc Repackaging and Shipping the ELx800 31 Repackaging the ELx800 and Its Accessories Refer to Figures 8 through 13 when repackaging the ELx800 1 Move the carrier to the home position 2 Turn off the unit and unplug the power supply 3 Wrap the shipping straps around the carrier as shown in Figure 8 Carrier Shipping straps Figure 8 Securing the carrier ELx800 Operator s Manual 32 Chapter 2 Installation 4 While holding the carrier carefully turn the instrument over and hook the shipping straps around the shaft Figure 9 5 Remove the four top cover mounting screws 6 Lift the instrument off the cover and turn it over Shipping straps NN Top cover Figure 9 Hooking the shipping straps around the shaft Bio Tek Instruments Inc Repackaging and Shipping the ELx800 33 7 Install the shipping block Place th
38. 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 Selecting Read Options on page 101 for more information 4 Place the plate in the carrier and then press the READ key to continue Bio Tek Instruments Inc Reading a Microplate 89 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 SELECT ASSAY NUMBER NAME HBS 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 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 Bio Tek s Extensions Define Reader Protocol software e Prompts enabled via UTIL 3 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
39. sensor cannot be interrupted I X axis carrier in out motor did not find the opto home sensor Note In cases where a sensor is not functioning the motor will drive the axis to its mechanical stop and generate substantial noise Bio Tek Instruments Inc Error Codes 143 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 e Defective or broken optical sensor e Sensor not connected e Y axis movement is limited so that the optical sensor cannot be interrupted Filter wheel did not find opto home sensor Probable Cause e Failed lamp Light beam not found During the X axis movement the light beam saturation transition max light to no light was never found during autocalibration Probable Causes e Loose belt loose motor pulley or defective motor drive causing the carrier to ignore movement instructions e Failed lamp e Loose filter wheel drive gear or defective motor controller PCB Filter wheel failed positional verify lamp failed e A failed lamp generates a 0302 error on power up ELx800 Operator s Manual 144 Chapter 6 Troubleshooting and Error Codes General Errors Continued Carrier X axis movement failed positional verify Motor X axis failed to get to the same position when moved a known number of ste
40. specify how replicates are mapped on the plate The currently selected Replication Direction appears on the top line of the display and the available options appear on the bottom DIRECTION ACROSS 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 Examples of mapping directions are shown on the next page ELx800 Operator s Manual 56 Chapter 3 Operation Examples of Mapping Directions Map Direction DOWN Rep Direction DOWN 1 2 03 34 5 j6 7 8 9 10 n jd smi smb5 sme J p j j sii STD5 SMP J p sm2 PC sme p j S sm2 jec p J pe T sno T ST T T eee me EE E T E e a srs ne j ST j j j Sre p Je o doo Oo sp qo o qo stoa sme d j T wf 19 19 o slpoe 06 10 8 29 0 NC NC SMP SMP SMP SMP SMP se 2 0J3 Ja s J 7 Js j9 po jn O o a gr gp oo qp ee ee ee m2 sr2 h CT sms srs J j j p j fJ SUPE SPD ad ee ee ee RSS enc a Fa ey ee ee ees Cee BGs Cs duo E Wl p cap n pene p eRe Fe ee e ee sme se wf
41. t 1 076 X 0 010 OD from 0 000 to 2 000 OD 405 nm Linearity t 1 076 from 0 000 to 2 000 OD 405 nm 3 076 from 2 000 to 3 000 OD 450 nm Repeatability STD 0 576 0 005 OD from 0 000 to 2 000 OD 405 nm e Optical specifications for the 340 to 400 nm range 50 second read in normal read mode Absorbance Measurement Range 0 000 to 3 000 OD Accuracy 2 0 0 010 OD from 0 000 to 2 000 OD 340 nm Linearity 2 576 from 0 000 to 2 000 OD 340 nm Repeatability STD X 1 576 0 005 OD from 0 000 to 2 000 OD 340 nm Optical specifications for the 400 to 750 nm range 30 second read in rapid read mode Absorbance Measurement Range 0 000 to 3 000 OD Accuracy 2 0 0 020 OD from 0 000 to 2 000 OD 405 nm Linearity 2 076 from 0 000 to 2 000 OD 405 nm Repeatability STD t 1 076 0 010 OD from 0 000 to 2 000 OD 405 nm ELx800 Operator s Manual 8 Chapter 1 Introduction e Optical specifications for the 340 to 400 nm range 30 second read Absorbance Measurement Range Accuracy Linearity Repeatability STD Narrow Beam NB Model Filters 0 000 to 3 000 OD 2 5 0 020 OD from 0 000 to 2 000 OD 340 nm 2 5 from 0 000 to 2 000 OD 340 nm 2 076 0 010 OD from 0 000 to 2 000 OD 340 nm 10 nm half bandwidth interference filters User accessible filter wheel Up to 5 filters may be installed on the instrument at one time Filters supplie
42. the OD on the linear scale for each calibrator e Automatic Use appropriately validated software and the curve fit that best fits the data e Manual Using log linear graph paper draw a smooth curve through the points not a straight line or point to point Treatment of Anomalous Points If any one point does not lie on the curve it can be removed If the absence of this point means that the curve has a shape dissimilar to that of the sample calibration curve or more than one point appears to be anomalous then the assay should be repeated Calculation of Autoantibody Levels in Controls and Samples Read the level of the anticardiolipin autoantibody in the controls and diluted samples directly from the calibration curve The control values should fall within the range given on the QC Certificate Note The standard values have been adjusted by a factor of 100 to account for a 1 100 sample dilution No further correction is required Assay Calibration The assays are calibrated against the Louisville reference LAPL GM 100 One GPL unit is defined as the cardiolipin binding activity of 1 ug ml of an affinity purified IgG anticardiolipin preparation from a standard serum The Louisville reference center recommends the following positive discrimination criteria according to the recommendation of the 2 4 International Anticardiolipin Workshop Criteria Range GPLU ml High Positive 80 Medium Positiv
43. using the Sigma powder 2 Ina beaker mix 50 ml of the PBS solution prepared from the Sigma powder with 10 mg of the B NADH powder and mix thoroughly This is the Sigma PBS Solution Bio Tek Instruments Inc Procedure 1 Liquid Testing 127 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 B NADH powder until the solution is at least at the lower end of this range Do not adjust if slightly high Carefully prepare a 75 Test Solution by diluting 15 ml of the 100 Test Solution gt If using the Sigma PBS Solution use 5 ml as the diluent gt 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 Carefully prepare a 50 Test Solution by diluting 10 ml of the 100 Test Solution gt If using the Sigma PBS Solution use 10 ml as the diluent gt 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 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
44. washing can be performed manually e Plate Reader Capable of measuring optical densities at 450 nm referenced on air e Distilled or Deionized Water This should be of the highest quality available e Calibrated Micropipettes For dispensing 1000 100 and 10 ul e Multichannel Pipette Recommended for dispensing 100 ul volumes of conjugate substrate and stop solution e Glass Plastic Tubes For sample dilution Quality Control and Results Quality Control For an assay to be valid all the following criteria must be met e Standards and the positive and negative controls must be included in each run e The values obtained for all the controls should be in the ranges specified on the OC Certificate e The curve shape should be similar to the standard curve shown on the QC Certificate e If the above criteria are not met the assay is invalid and the test should be repeated Calculate Mean Optical Densities for assays run in duplicate only For each standard control and sample calculate the mean OD of the duplicate readings The user must verify that the percentage coefficient of variation 76 CV for each duplicate OD is less than 15 0 ELx800 Operator s Manual 182 Appendix E Instructions for Programming a New Assay Plot Calibration Curve The calibration curve can be plotted either automatically or manually as follows by plotting the anticardiolipin autoantibody concentration on the log scale against
45. wavelength gt 340 UV model only 405 415 450 490 515 540 550 562 570 590 595 600 620 630 650 660 690 750 e Terasaki plate adapter for 60 72 and 96 well Terasaki plates for ELx800NB and ELx800RNB PN 7330531 e Absorbance Test Plate PN 7260522 e Epson LX 300 printer or equivalent 110V only PN 97145 e Sheet feeder for Epson LX 300 PN 97144 e HP Deskjet printer PN 97152 e KC4 Software gt KC4 Reader Control and Data Analysis Software PN 5290512 gt KC4 Signature 21 CFR Part 11 Compliant Version PN 5290510 gt KC4 Signature Upgrade from v2 7 or lower PN 5290511 gt KC4 Signature Upgrade from v3 0 PN 5290508 gt KC4 Upgrade from v2 7 or lower PN 5290514 e KCjunior Software gt KCjunior Software Package PN 5270501 gt KCjunior Upgrade PN 5270508 e Delta Soft Macintosh Software PN 3070145 e ELx800 Qualification Package PN 7330538 ELx800 Operator s Manual 6 Chapter 1 Introduction Specifications Standard Model Wavelength Range 400 to 750 nm Filters 10 nm half bandwidth interference filters User accessible filter wheel Up to 5 filters may be installed on the instrument at one time Filters supplied 405 nm 450 nm 490 nm and 630 nm e The following specifications apply to 96 well flat or round bottom plates single wavelength measurements with a 50 second read normal read mode Absorbance Measurement 0 000 to 3 000 OD Range Accuracy
46. 153 Problems ue dida ue pd ox vx datu aab e ux tva au afta esee Uv Vnd 154 Getting Started With KC4 L uuo ssec x a RE Ie a n o S R at 154 150 Appendix A Controlling the Reader With KCjunior or KC4 Overview Using KCjunior or KC4 software provides the user even more power and flexibility For example complex assays using features not available through the front panel may be defined and run This Appendix provides instructions for controlling the reader with Bio Tek s KCjunior or KC4 software packages Bio Tek Instruments Inc Controlling the Reader With KCjunior 151 Controlling the Reader With KCjunior The ELx800 can be operated using a computer running Bio Tek s KCjunior software Follow the steps below 1 ee Power off the computer and the reader Connect the serial cable PN 75053 between the two machines Power up both machines Install KCjunior on the computer s hard drive Once installed start KCjunior Select Setup then Reader1 To select the reader and define the communications parameters choose the following setup parameters Reader ELx800 EL800 or ELx800UV EL800UV or ELx800NB EL800NB Com Port COMI or COM the serial port used for the RS 232 cable connection gt Baud Rate 2400 4800 or 9600 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 Clic
47. 23 Parallel port printer port Power supply jack Figure 6 Connectors for printer parallel computer serial and power supply ELx800 Operator s Manual 24 Chapter 2 Installation Setting Up the Serial Port for Communications With Other Devices Before serial communication can be initiated between the ELx800 and another device such as a host PC running Bio Tek s KCjunior or KC4 software the communication parameters must match between the devices The ELx800 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 ELx800 software Appendix A contains information on required protocols for computer control of the reader Attaching the Cable Power down the computer and the ELx800 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 Ensure that the ELx800 and the computer are operating with the same communications settings Setting Communication Parameters The reader s default communication parameters are 9600 Baud Rate 8 Data Bits 2 Stop Bits No Parity The baud rate can be changed to 1200 or 2400 bps if necessary The Data Bits Stop Bits a
48. 2400 4800 or 9600 must match the baud rate on the reader Data Bits 8 Parity None gt Stop Bits 2 Click the Current Reader button to attempt to establish communications with the reader using the currently defined communication parameters If the test passes click OK to save the settings and close the dialog box If the test fails KC4 will provide appropriate instructions for resolving any problems See also the Problems section below To learn more about KC4 and how to define assays and read plates refer to the KC4 User s Manual ELx800 Operator s Manual 154 Appendix A Controlling the Reader With KCjunior or KC4 Problems 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 and is not a Null cable If this is suspected add another Null and try again If an Incorrect Reader Model Connected dialog box is displayed click OK to clear the screen and select System Readers Available Readers Verify that the reader selected is correct 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 2 10 Select Data New Plate If prompted to select a p
49. 55 ASSAY 55 Figure 37 Assay Detail Report Sheet 2 of 2 Bio Tek Instruments Inc Appendix D Comparison of the ELx800 and the ELSOO The following chart compares the features on the ELx800 and EL800 microplate readers ingle and dual wavelength reading The EL800 version can be computer controlled and will function as an ELx800 O1 All ELx800 optical specifications apply to the ELx800 172 Appendix D Comparison of the ELx800 and the EL800 172 Bio Tek Instruments Inc Appendix E Instructions for Programming a New Assay This appendix provides two examples of assay kit instructions and step by step directions for programming each assay The appendix includes two sample assays one with a ratio transformation calculation and a POS NEG cutoff determination and another with a standard curve These examples are based on real assays however the results have not been verified on any clinical kits The kit instructions are provided so that users can see how it is possible to translate the kit wording into an ELx800 assay program For clarity only the user menu choices from the reader screens are shown Refer to Chapter 3 for details Important The ELx800 user is responsible for programming the reader properly according to their specific kit instructions and for verifying that the calculations are performed correctly Sample ANA Screen Enzyme Immunoassay Kit ssss 174 P
50. BIO TEK ELx800 late Reader e um Ps zt o Q V F a JT o c aa lt O Ge OK Creating Solutions for the Future of Science 7331000 Rev N2 July 2005 This Manual Update contains changes to the ELx800 Operator s Manual These changes will be incorporated in the next full revision Rev O of the Manual Page 43 System Startup Addition software installed Not all features of the software discussed D Important Some ELx800 readers have custom programmed in this Operator s Manual are available on custom instruments Please contact Bio Tek s Technical Assistance Center at 800 242 4685 if you have any questions about the software on your reader Page 119 Liquid Test 1 Recommendation After pipetting the diluted test solution into the microplate step 3 and before reading the plate we strongly recommend shaking the plate 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 GJSBIO TEK BIO TEK INSTRUMENTS INC P O BOX 998 HIGHLAND PARK WINOOSKI VERMONT 05404 USA COPYRIGHT 2005 TEL 888 451 5171 FAX 802
51. Calls on the next page for information on how calls are assigned Bio Tek Instruments Inc Define Method Map Formula and Curve 79 Positive Negative Calls for Cutoff After the greyzone is defined calls for the sample wells Positive NEGative EQUIVocal must be defined Select POS or NEG to select the call that will be assigned to samples greater than the cutoff value plus the greyzone 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 lt CUTOFF CUTOFF GREYZONE Examples 1 The cutoff between negative and positive calls should be calculated as the average of the negative controls plus the OD value of 0 500 Samples greater than the cutoff should be labeled 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 x40 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 POSifthe sample is greater than 1 500 NEGifthe sample is less than 1 500 ELx800 Operator s Manual 80 Chapter 3 Operation 2 Fora quantitative assay samples with OD values greater than the STD2 mean plus
52. 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 70 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 MAP before creating blank or control validation formulas Blank Control validation can be performed on individual replicates BLK PC oron the group mean BLK x NC x Examples If an assay protocol states that gt Each blank well on a plate should have an OD of less than 0 050 the formula is BLK lt 0 050 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 000ANDPC
53. 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 Directive 98 79 EC In Vitro Diagnostics e Product registration with competent authorities e Traceability to the U S National Institute of Standards and Technology NIST Optical density measurements and if equipped incubator temperature readings are traceable to NIST Specific data for a particular serial number is available on request from Bio Tek Instruments See page xi for contact information ELx800 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 equipment is operated in a commercial environment Like all similar equipment this equipment generates uses and can radiate radio frequency energy and if not installed and used in accordance with the instruction manual may cause harmful interference to radio communications Operation of this equipment in a residential area is likely to cause interferen
54. O DOWN DOWN A01 BLANK MAP AIR NUMBER STDS 05 NUMBER STD REPLICATES 01 CONCENTRATIONS STD1 6 25 STD2 12 5 STD3 25 STD4 50 STD5 100 NUMBER CTLS 02 CONTROL 1 PC CONTROL 2 NC NUMBER OF REPLICATES PC 01 NC 01 SAMPLES 89 SAMPLE REPLICATES 01 Bio Tek Instruments Inc STEP To define a cutoff formula for Positive and Negative calls Curve 4P 185 COMMENTS Kit instructions specify that samples with concentration values greater than 11 should appear as positive The ELx800 software calculates the cutoff based on absorbance value or transformed value see the previous example and cannot calculate based on concentration The technician must make the positive or negative determination visually based on the calculated concentration results As a general guideline choose linear if you expect a straight line result Choose 4P for all others unless otherwise specified by the kit instructions The reader will automatically calculate the concentrations of the samples when the assay is run ELx800 Operator s Manual 186 Appendix E Instructions for Programming a New Assay Bio Tek Instruments Inc
55. Plate is correctly seated in the microplate carrier with A1 in the upper left corner 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 Bio Tek representative or Bio Tek Technical Support 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 is shown that the device is accurate at these OD values then by definition it is 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 test review the following possible problems and solutions gt Check the 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 Bio Tek Instruments Inc Qualification Procedures 111 gt Ensure that the filter calibration values entered are the same as those on the Test Plate data sheet 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 Bio Te
56. RIEKRASVTURUEPA SURE UEEI QC E EdErge e 168 Assay Detail Report Sheet 1 of 2 csse nnne 169 Assay Detail Report Sheet 2 of 2 ccccccccsccssseeseeeeseseneseenaees 170 List of Tables Table 1 Recommended Test Schedule i eene r eterna tre e nre hr x nex annt 105 Table 2 Typical Enzyme Substrate Combinations and Stopping Solutions 116 Table 3 Test Tube DIIUEIDITS ecce te rape tem eee den detener ei rel ree nod eoe 121 Table 4 PBS 10X Concentrate SO ULEDIOFO roo rn erre nnm tn ne 126 Bio Tek Instruments Inc Contact Information xi Contact Information Bio Tek Instruments Inc Highland Park P O Box 998 Winooski Vermont 05404 0998 USA Customer Service and Sales Internet Phone Fax E Mail Service TAC Phone Fax E Mail www biotek com 888 451 5171 toll free in the U S 802 655 4040 outside the U S 802 655 7941 customercare biotek com 800 242 4685 toll free in the U S 802 655 4740 outside the U S 802 655 3399 tac biotek com European Coordination Center Bio Tek Instruments GmbH Kocherwaldstrasse 34 D 74177 Bad Friedrichshall Germany Internet Phone Fax E Mail www biotek de 49 0 7136 9680 49 0 7136 968 111 info biotek de ELx800 Operator s Manual xii Preface Document Conventions This manual uses the following typographic conventions Example Description A This icon calls attention to important safety
57. Ra d DRE CR Scd 24 Setting Communication Parareters ccees uoxaukx i nn nnne xn RR n RR X RR ER RR 24 Installing Additional Filters 25 are hirta Ra n E XEDRE REA ERA AX EX EIER EUER 26 Bio Tek Instruments Inc Contents v Repackaging and Shipping the ELX800 orit sin int hen tentant akku 30 Before Repackaging the Instrument sees nennen nnn 30 Repackaging the ELx800 and Its Accessories sssssrsserrrrrsrrrrrssrrrrrsnnrnn 31 Preparing the Shipping Container siaisiscesewiasewsadewrssdor cia cudusaanassaaveesnnnes 37 Chapter 3 ObDSFOLIOD aissxssimivansEn tiu mcFpE E pu ER Ein iD aa aaia E ad 39 ELX800 Front Panel ixssenisuasxecrot c Ue uk tex c a Ee biSe pe n NEA ER UC EA EIE Ded 40 e allo M 42 Recommendations for Achieving Optimum Performance eeeeeeeeee 42 System Startup useoxessax EA d RR UO QOO EO P AG b RA OL Rat e RH DV RON OR la 43 Mali MENU is cts Sees E Eee mE E b E iL UE EE ere ME 44 DENE cp 46 Programming INGUG P M 47 Selecting an Assay to Define sssssseesseeesseeeen enne nennen nna nnn nnne 48 Editing the Assay INGTUbos aspi daran p i ea ata e irat clc kh E Y eda 48 Define Method Map Formula and Curve iiis rcr ok ki 49 Defining METHOD cesa duvak os v ACEE Sa EE ExRENEF a ERAI FE ar oo dU REOR VADE
58. Reading from the main menu The Reading parameters dialog will appear Reading ht m Reading Type m Reader c EnB ELx800 on COM C Kinet diis Kinetic Scanning Filters Meas Filter 405 Pre Read Blank Plate C Refr Filter Monitor Wells Options Plate Type 96 WELL PLATE Size 8x12 First well fat Last well H12 Cancel Help Figure 30 Reading Parameters dialog Select a Reading Type of End Point or Kinetic Define the wavelength s at which the plate will be read Select a Plate Type Define other reading parameters as necessary Click the Help button for assistance When complete click OK Select Data Read Plate The Plate Reading dialog will appear Enter any comments 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 ELx800 Operator s Manual 162 Appendix B Using 384 Well Geometry Bio Tek Instruments Inc Appendix C Report Format This appendix contains examples of reports that can be generated and or printed from the ELx800 Refer to Printing Reports in Chapter 3 for details on how to print these reports In addition an Assay List Assay Definition Map and Resu
59. TI MEE A ERR Gi 10 PHONG SUP DOM yerigen seresa rT E usas ax PEVET VIQR MITES A LETEVVNE REA TOYA EM OS VAT EAR dose 10 Electronic COMMUDICATION ut iieovas eros d euo read deret ed suere Pear sa rrr Ju D RA 11 Facsimile Sup DOR estado c ar B lote ol P Vb EU E 11 Written Communication sirrini nri nU Pu rr Rae YA RADAR DX USER aai ne aa 11 Shipping Address oc sita x aaa a E RATA Fa A a a Cua YR aban 11 Chapter 2 Installation ssscccissanisnistienscssucsesstecesswecewsvcusstccsassnaueniweneuce 13 Unpacking and Inspecting the EEX8001 v cio ici dd OR PROC C ER CE EGRE EAE 14 Unpacking the Instrument and Its Accessories sssssrrrrrrrrrrrssrrrrrsrrrnne 15 Setting Up the ELXSOO c LassxtenvaseuniVads zu uar E rab Dua ea dea Epic xw cet du Ua d abr Dur valeas 19 Operating EDwvIEOnbmenE icine adaware vena eina C EOS de cx edo de Ma qe ea Fa n 19 Electrical Connections oo evene tous Krise diss de ievveosAdu ek onesies wenn Fei a ta nU ex tos 19 Power Up and System Test soie cave cedi aV e C an on EO ta kl c cal Vc e nai at 20 ELX800 Math Menu 2d eder ER cedex Ox x veneta siete dide hid sad te atlas he eames 20 Configuring Global Default Options ceeeseeeeeeeennnen nnne nnn 21 Connecting a Printer to the ELX800 usoosuset seuewekagas ave qusaue aaa xorkaxea saute qaas 22 Setting Up the Serial Port for Communications With Other Devices 24 Attaching the Gables uiv eor EoX ek tetti deni ces di v owe ed Va Ya ron ERU
60. TUA TR V VE VOX VA E E 49 Defining MAP sssrin en ed reda ebaxcd toes ob ad exu etd Eovaddatra da bn Rd D RUNI 53 Denning FORMULA srecne nena EEE E Mx ERN ERU E FERE A uM DAT paro d E ME 69 Defining GW c EE 81 RSS el MICEDDIES usa diuskadE ERE AM Rd CERA ODE T den adnan ch KADER CRM E ERR 88 Selecting am Assay to TUN usa dacesaca Vn ORAL SG GERA x Exclu dd pa ic n T nn OA RR 89 Printing Re POs iss scares rests divides vaga ea usn A RU ra E Qu TU EP UI AV KOnElda cad rue Cp edd lbid dau 93 Editing Standard Outliers iaxeuis sawtiaza iasburiubiwdakiwisgas odas d qax ndn nies Lessnanianaas 94 PMP RESHIES ciii eru expe caa pa ao Ead tac da nasce raa ake aa venia s Fa ipaa x waa V d arida 96 Using the Utility ODUOHS ied rete qua tele ayuu E dac raus E pakus uus v eC yat eu rEOp eae pb ERO E uds 97 Setting The Date and TIME usas censausx RE RR ERAS d aiu NY GRE EA EAR ER a DWE 97 Viewing Editing the Filter Tabl iisiciswcssccvset cuts rrr rry hh rta hara raa 98 Specifying Data Output and Reporting Options ccccccceeseeeeeeeeeeeeeeeee ees 99 Selecting Read RE Le 1 TTC TTL UU IIT 101 ELx800 Operator s Manual vi Preface Chapter 4 Performance Verification and IQ OQ PQ Procedures 103 Recommendations for Achieving Optimum Performance eeseeeee 104 Recommended Test Schedule rre ny nr e ro re 105 Qualification Procedures iioii nain e Ra ANE ERR YRARR RETE RR ARIRET NERA
61. Test 1 Preparation of Dilutions 1 Set up a rack containing 10 tubes numbered consecutively 2 Ifyou have not already done so prepare concentrated stock Test Solution A or B Refer to Table 3 when executing steps 3 and 4 Table 3 Test Tube Dilutions Tube Number HORE MN ae EIU Volume of Concentrated 20 18 16 14 12 10 Solution ml Volume of 0 05 Tween 10 12 14 Ed Solution ml Absorbance expected if x E 1 2 1 0 0 a concentrated solution is 2 0 at 200 ul 3 Create a percentage dilution series beginning with 100 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 3 ELx800 Operator s Manual 122 Chapter 4 Performance Verification and IQ OQ PQ Procedures 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 3 as a model for calculating the expected absorbances of a series of dilutions given a different absorbance of the original solution Read Plate 1 Read the
62. VR DA 106 System TSE chou stairs ein terc WR PI ER xa EE rV eR LOL VIP MN a EET E 107 Checksum TSE a T 109 ADSOFDANCE Plate SE s resumes enirn et a santas PA ee cesar cx bac onda ions 110 Entering the Absorbance Plate Specifications csse 113 Test Falliles xcoidtusc ERE UD RE Ferd ids cra OD AREE UD NER FCU dd 115 Liquid Testing suisse ce oic ae Y ear acd d E dolar OL EE C OR Vr T C RO a 116 Stock Solution FOrrmulatiOll 22v ioa kao xen a neo nadie Ee ERE Er IEEE pP e e uei ArEd 117 Liguid Test 1 ivive cksus ib eaden Save VERTES uA SE VE Fa EVARRR VITIS A ZA ERA AA E CREMA NA RR 119 Lice Test 2 ato aeea 2d dade EEA Qvae sane lera eae ema eee Paus ea vo Vll aoo 121 Liquid Test 3 for UV Models ONLY sinwisusesarwere aan c b ae aD EROGO dc 125 Chapter 5 Maintenance and Decontamination 129 Haisi is as tee eM 130 Routine Cleaning Procedure use teda ceni ien ce pan e bx wae rel CO e Re er edd 130 PUBPOSG m D EET 130 Tools and Supplies scies Iu Yd vue kn RTI E aks Gaul ovn nk ERR e Rd ORO E ON B d EROR RT 131 Procede iii a Decree ER 131 Replacing the Bulb 30 sackeevewekaw v ped na ove en oe wc Orta Ex xo ies aa e potions lv ina 132 Decontamination Procedure s ssssssssssssrrsnrrrrnnnsnsnenunrnnnnnrnnnnnnnnnnnunnnnnnnnnnn 135 LOGS and SUPphe Sisena A EA EA E eei dd 136 lise MPH RE 136 Bio Tek Instruments Inc Contents vii Chapter 6 Troubleshooting and Error Codes
63. 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 gt POS if the sample is greater than 2 200 gt NEG if the sample is less than 1 800 Bio Tek Instruments Inc Define Method Map Formula and Curve 81 Defining CURVE Note This feature is not available in the EL800 version see Appendix D to compare the EL800 with the ELx800 To define curve fitting parameters for an assay 1 Start at the Main Menu and select DEFINE 2 Select the assay and then press ENTER The DEFINE option screen will appear 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 Note These screens are displayed on the ELx800 in the order in which they appear in the assay If a closed variable i e an element of the assay definition that you cannot access or modify is being used in the assay the entry screen is omitted Curve Fit Type The EL
64. age Contents ree Ee t cusa eU opesducta isa abd ee ep Av D Optional Accessories GL Jigs alee oic eck da tad S Een AR ROVER DE Ca AD Ad SPEGITIGATIONS eret osd eee sod iea cov edes sr to eee ns eot as TOU EY Ese Standard Models uode sere ree inner da ee rea nea OE vede tua Ultraviolet UV Model c sese ememem enn nn Narrow Beam NB Model ccsessse meme nnn technical SUPPO tunni verre n oat e poa a cans e o ROC ERR 8 LER Rc a 2 Chapter 1 Introduction Introducing the ELx800 Automated Microplate Reader The ELx800 Automated Microplate Reader is a single channel reader assay system designed to automatically perform endpoint analysis for a variety of ELISA based applications The reader can measure the optical density of solutions in 6 12 24 48 or 96 well microplates The NB reader models can measure 384 well microplates and 60 72 96 well Terasaki trays as well as 6 12 24 48 or 96 well plates The reader features superior optical specifications with an extended dynamic range of up to 3 000 absorbance units in some read modes The wavelength range is from 400 nm to 750 nm UV instruments have an extended range from 340 nm to 750 nm Kinetic analysis can be performed using computer control e g via KC4 or KCjunior running ona host PC 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 charac
65. ance to the next replicate gt If 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 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 ELx800 Operator s Manual 96 Chapter 3 Operation Printing Results After the assay is selected and standard outliers are edited if necessary the results report can be printed PRINT RESULT SIS Y ES N 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 ASSAY NUMBER NAME HBS G 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 ASSAY NUMBER NAME HBS G e Press Options to cyc
66. ances to the next screen in a series Pressing Previous Screen generally saves the current screen settings and returns control to the screen most previously viewed 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 Press a number to enter it in a data entry screen Press Shift a number for example Shift 7c to enter the letter then press Options to advance through the alphabet ELx800 Operator s Manual 42 Chapter 3 Operation Overview IMPORTANT Do not turn on the instrument until the carrier shipping block has been removed The ELx800 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 ELx800 Recommendations for Achieving Optimum Performance Microplates should be perfectly clean and free of dust or bottom scratches Use new microplates from sealed packages Do n
67. ap 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 ELx800 Operator s Manual 60 Chapter 3 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 ENTER BLANKING 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 e Press ENTER to continue Number of Blanks The NUMBER OF BLANKS field allows you to enter the number of blank wells in the assay This entry screen is only displayed when Full Column or Row blank maps is selected Any previously defined value is displayed ENTER NUMBER O F BLANKS e Use the numeric keys to enter the number of blanks The range is 0 to 48 e Use the CLEAR key to clear the NUMBER OF BLANKS value from the display e Press ENTER to continue Bio Tek Instruments Inc Define Method Map Formula and Curve 61 Blank Location The BLANK LOCATION screen allows you to define where the blank well or wells occur on the microplate
68. avelength assay 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 When selections are completed the display returns to the SELECT UTILITY OPTION menu ELx800 Operator s Manual 102 Chapter 3 Operation Bio Tek Instruments Inc Chapter 4 Performance Verification and IQ OQ PQ Procedures This chapter discusses the tasks and procedures necessary for verifying and qualifying instrument performance on an ongoing basis A convenient Recommended Test Schedule arranges tasks into Installation Operation and Performance Qualification categories Recommendations for Achieving Optimum Performance Recommended Test Schedule ccccsccssseesseeeeeeeseeeesseeeeeenseneeas Qualification Procedubes oen tu eve ei eoa Cr c vie Pe ERRARE COR a System TeSUL oco e dra vdune T EE RE vds aul detvves TV VY Ud a eset CHECKSUM DOSE cosas csis ek vae ce exacta hae ated eap n Ea oZ nad Nas Absorbance Plate Peskosucuscenexxcor siavavadsansaeweene dws sinus dax Fu EE M Entering the Absorbance Test Plate Specifications Test Fallill G5 veri icut duae ovile trae Cava a o doy sod e D PR xr LT OREA BIG RES CIN G ead omae rant xg Dr e aca a RC d dar daa e e d Stock Solution Formulation sseeseeeee mn INTu EUIS BII aA epe Pr CETT LIGUIG TESU P
69. ay STEP SAMPLES 91 SAMPLE REPLICATES 01 Use the MAP and MATH keys to create the control validation formula FORMULA VAL CONTROL PC x gt 1 200 NO OF REPLICATES 01 NO x lt 0 300 NO OF REPLICATES 01 To create the plate transformation for CALC divide the ANA result by a cutoff standard FORMULA MORE TRANS VAR SCOPE VARIABLE SMP or OD FORMULA CTL1 X TRANS FORMULA OD TVAR 10 To define a cutoff formula for Positive and Negative calls FORMULA CUTOFF 10 0 GREYZONE 00 SAMPLE gt CUTOFF 00 POS COMMENTS User defined recommendation is to fill the entire plate When defining control validation formulas use PC to indicate the criterion for each of the PC replicates and PC x to indicate the average of the Positive Control replicates Select OD to advance to the formula definition screen and define the transformation variable Tvar Defining TRANS VAR CTL1 X isolates the OD value for CTL1 X for use in transformation Converts all OD values on plate to ANA Result per kit insert instructions Bio Tek Instruments Inc 179 Sample Human Anticardiolipin IgG Enzyme Immunoassay Kit Standard Curve and Cutoff Intended Use This assay is intended for the in vitro measurement of IgG anticardiolipin antibodies in human serum as an aid in the diagnosis of antiphospholipid syndrome APS Sufficient materials are supplied to allow a maximum o
70. azard 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 contact the Bio Tek Service Department Do not soak the keypad this will cause damage Important Turn off and unplug the instrument for all decontamination and cleaning operations 1 Turn off and unplug the instrument 2 Prepare an aqueous solution of 0 5 sodium hypochlorite NaClO or bleach As an alternative 70 isopropyl alcohol may be used if the effects of bleach are a concern gt Be sure to check the percent NaClO of the bleach you are using this information is printed on the side of the bottle Commercial bleach is typically 10 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 Bio Tek Instruments Inc Decontamination Procedure 137 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 d
71. 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 Note Uses linear axis type only 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 Bio Tek Instruments Inc Define Method Map Formula and Curve 83 Edit Standard Outliers This screen allows you to enable or suppress the editing of standard outliers 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 is displayed EDIT S T D OUTLIERS MANUAL NONE MANUAL e Select NONE to suppress the EDIT STANDARD OUTLIERS capability for this assay e 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 After the assay is run and reports are generated press REPORT from the Main Me
72. ce in which case the user will be required to correct the interference at his her 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 emet 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 e UL 61010A 1 ist Edition Safety Requirements for Electrical Equipment for Measurement Control and Laboratory Use Part 1 General Requirements International EN 61010 1 2001 Safety Requirements for Electrical Equipment for Measurement Control and Laboratory Use Part 1 General Requirements Bio Tek Instruments Inc Safety Symbols Safety Symbols xxi Some of the following symbols will appear on the instrument ONS d ELx800 Operator s Manual Alternating current Courant alternatif Wechselstrom Corriente
73. cted performance of the batch Cardiolipin Coated Wells 12 break apart 8 well strips coated with bovine cardiolipin antigen The plate is packaged in a re sealable foil bag containing two desiccant pouches Type II Sample Diluent 2 bottles containing 50 ml of buffer for sample dilution Colored yellow ready to use Type II Wash Buffer 20x Concentrate 1 bottle containing 50 ml of a 20 fold concentrated buffer for washing the wells Cardiolipin IgG Standards 5 bottles each containing 1 2 ml of diluted human serum with the following concentrations of anticardiolipin autoantibody 100 50 25 12 5 6 25 GPL U ml Ready to use The standard set is calibrated against the Louisville APL reference preparation Cardiolipin IgG Positive Control 1 bottle containing 1 2 ml of diluted human serum The expected value is given on the QC certificate Ready to use Cardiolipin Negative Control 1 bottle containing 1 2 ml of diluted human serum The expected value is given on the QC certificate Ready to use Cardiolipin IgG Conjugate 1 bottle containing 12 ml of purified peroxidase labeled antibody Colored red ready to use TMB Substrate 1 bottle containing 14 ml TMB substrate Ready to use Stop Solution 1 bottle containing 14 ml of 3M phosphoric acid Ready to use Bio Tek Instruments Inc 181 Additional Materials and Equipment Not Supplied e Automatic Microplate Plate Washer This is recommended however plate
74. ctions for Programming a New Assay 173 Sample ANA Screen Enzyme Immunoassay Kit s ssssssssssrrrrrrrrrressssrrrrrrrrns 174 Intended USE nersini aaia Ek XE REX KR KR ERN RERK KNEE EEYEERNA EAE ONE REDE 174 Background cci oW a VERI YEAR EREIEXWEXA REN IRAE adie woe 174 Principle Of the AsSay scii er rea Ra Dey WEAR E ERRER RR EXRRE ERE NAR AREIR RA LAN 174 Pra CE rile e nU 175 Quality Control and Results 2 over cxi rna yan rnt ree cnra ke n Eco ree ce 176 Expected Val eSa dicta si cedi kv i paria died chinese E o za Sa OE Y e ER v Ca Or a D eeuas 176 Programming the ANA Screen Enzyme Immunoassay Kit sssssss 177 Sample Human Anticardiolipin IgG Enzyme Immunoassay Kit 179 Intended USC iccirco ke an Eu x oc ors e IMS LA Schaal 179 Background eisernen peka RARE DI d RE RR E A Sa IRE ani gexaupdLulabtS adds Paid eva DpEs bass 179 Principle of the ASSAY asia vsawerkut es sioe 9x e Exo rix pd Pone rapit kenn aa dE a FR Yd aa 179 Materials s T M 180 Ouality Control and Results ier adnan s dia d e a i Ox obe o e vale ka Ge n 181 Expected Values iere FExa ER ERRARE e ERARARWRREA TED CY REYER KRRRR EE TAA wees 183 Programming the Human Anticardiolipin IgG Enzyme Immunoassay Kit 184 Bio Tek Instruments Inc Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Supply Figure 7 Figure 8 Figure 9 Figure 10 Figure 11 Figure 12
75. curve was originally generated on e g the curve for Assay 53 cannot be applied to samples on a plate to be run in Assay 51 To reuse a standard curve you must first program an assay in positions 31 through 55 and then run the assay During the defining process you will be prompted to enter the number of standards the number of standard replicates and the standard concentrations The following screen will appear after these prompts REUSIE STANDARD CURVE YES Y ES N O After the assay has been run the results have been calculated and the reports have been generated the reader will prompt if this standard curve should be stored in memory The following display will appear SAVE STANDARD CURVE Y E Y ES N O Select YES to store the curve for use at a later time The next time a plate is to be read using this assay the instrument will prompt if there are standards on the plate Select NO to discard the curve Bio Tek Instruments Inc Define Method Map Formula and Curve 65 STANDARDS If YES is chosen a new standard curve will be generated The plate map is not changed If Prompt for Sample ID is enabled in the UTIL section you will be prompted to enter the number of samples See Other Utility Options in Chapter 3 for more information on the UTIL options If NO is chosen the stored standard curve will be used If Auto mapping had been used to originally map the standards blanks controls and sa
76. d 405 nm 450 nm 490 nm and 630 nm The following specifications apply to 96 well flat or round bottom plates with a 50 second read normal read mode Absorbance Measurement Range Accuracy Linearity Repeatability STD 0 000 to 3 000 OD 1 0 0 010 OD from 0 000 to 2 000 OD 405 nm 1 076 from 0 000 to 2 000 OD 405 nm 3 076 from 2 000 to 3 000 OD 450 nm 0 576 0 005 OD from 0 to 2 000 OD 405 nm Bio Tek Instruments Inc Specifications 9 The following specifications apply to 96 well flat or round bottom plates with a 30 second read rapid read mode Absorbance Measurement Range 0 000 to 3 000 OD Accuracy 2 0 0 020 OD from 0 000 to 2 000 OD 405 nm Linearity 2 0 from 0 000 to 2 000 OD 405 nm Repeatability STD 1 0 0 010 OD from 0 000 to 2 000 OD 405 nm The following specifications apply to a 384 well plate The minimum volume of fluid in a well to obtain the specified performance is 80 microliters The specifications apply to the dual wavelength mode of read only Absorbance Measurement Range 0 000 to 3 000 OD Normal Read Mode Dual wavelength 4 minutes 45 seconds Accuracy 2 0 0 020 OD from 0 000 to 2 000 OD 405 nm Linearity 2 5 from 0 000 to 2 000 OD 405 nm Repeatability STD t 1 576 0 010 OD from 0 000 to 2 000 OD 405 nm Rapid Read Mode Dual wavelength 3 minutes 25 seconds Accuracy 2 576 0 020 OD from 0
77. d accessories that were furnished with the standard models THIS WARRANTY CONTAINS THE ENTIRE OBLIGATION OF BIO TEK INSTRUMENTS INC AND NO OTHER WARRANTIES EXPRESSED IMPLIED OR STATUTORY ARE GIVEN PURCHASER AGREES TO ASSUME ALL LIABILITY FOR ANY DAMAGES AND OR BODILY INJURY OR DEATH THAT MAY RESULT FROM THE USE OR MISUSE OF ANY EQUIPMENT OR INSTRUMENT BY THE PURCHASER HIS EMPLOYEES AGENTS OR CUSTOMERS OTHER THAN THE EXPRESS WARRANTY CONTAINED HEREIN WE SHALL NOT BE RESPONSIBLE FOR ANY DIRECT OR CONSEQUENTIAL DAMAGES OF ANY KIND THIS WARRANTY SHALL NOT BE CHANGED OR MODIFIED IN ANY WAY WITHOUT THE EXPRESS WRITTEN PERMISSION OF AN OFFICER OF BIO TEK INSTRUMENTS INC ELx800 Operator s Manual XXiv Preface Registration Card Once the instrument has been set up and is running successfully please take a moment to fill out and mail the postage paid Warranty Registration card By sending in the registration card you will be assured of receiving prompt information on product enhancements Bio Tek Instruments Inc Chapter 1 Introduction This chapter introduces the ELx800 Automated Microplate Reader and describes its hardware and software features Also included is contact information if technical assistance is needed Introducing the ELx800 Automated Microplate Reader Hardware Features aori ese pe nee veers EATE aa de i A TA NETERA Software Features es viu Ere opbr eR EA DENE AALER AARAA LAARA Pack
78. data acquisition and reduction parameters for a new assay or modify previously defined assays stored in memory I Start at the Main Menu and select DEFINE to display the SELECT ASSAY NUMBER screen SELECT ASSAY NUMBER NAME HBS AG 1 Select an assay to define or modify and then press ENTER See Selecting an Assay to Define on page 48 for detailed instructions The EDIT ASSAY NAME screen will appear If you are modifying selecting a PANEL assay 99 see page 85 for instructions NAME HBS AG 1 Optional Edit the assay name and then press ENTER See Editing the Assay Name on page 48 for detailed instructions The DEFINE menu will appear FORMULA CURVE The following options are available within the DEFINE menu METHOD Define the wavelength type single or dual wavelength s and plate geometry page 49 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 53 FORMULA Define cutoff transformation and validation formulas Create variables to be used within formulas page 69 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 81 Bio Tek Instruments Inc Define 47 Programming Note Assays with certain c
79. e gt 20 80 Low Positive 2 10 lt 20 Bio Tek Instruments Inc Results Interpretation The association between low positive levels of anticardiolipin antibodies and clinical findings is unclear 183 Normal population studies indicate that there is a higher prevalence of IgM positives in the normal population than IgG 9 4 and 6 5 respectively In normal pregnancy the levels are higher still at 17 0 IgM and 10 6 IgG Expected Values The normal range was determined on serum from 102 normal adult blood donors The ranges below are provided as a guide only ELISA assays are very sensitive and capable of detecting small differences in sample populations It is recommended that each laboratory determine its own normal range based on the population techniques and equipment employed IgG Anticardiolipin 11 GPLU ml Negative result 11 GPLU ml Positive result ELx800 Operator s Manual 184 Appendix E Instructions for Programming a New Assay Programming the Human Anticardiolipin IgG Enzyme Immunoassay Kit Standard Curve and Cutoff From the Main Menu press DEFINE Select the assay name and edit the name if desired At the define menu follow steps below STEP COMMENTS To program the reading method press METHOD WAVELENGTH Dual or Single Single MEASURE Wavelength s to use 450 PLATE TYPE 96 To program the plate map press MAP AUT
80. e If running a custom assay such as one that was created using Extensions software typical prompts might include gt The number of samples Standard concentrations gt Assay ID Fill pattern gt Blank method First well location gt Replicate count for each well type Wavelength mode Report preferences etc ELx800 Operator s Manual 90 Chapter 3 Operation Enter Number of Samples If the ENTER NUMBER OF SAMPLES prompt is presented indicate the number of sample groups on the plate The number of sample replicates is typically predefined in the assay but if this is a custom assay you may also be prompted to enter the replicate count ENTER NUMBER O F 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 than 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 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 ov
81. e The ELx800 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 ELx800 Front Panel in Chapter 3 for additional instructions Bio Tek Instruments Inc Setting Up the ELx800 21 Configuring Global Default Options The ELx800 contains several global configurable options such as date and time report output and plate reading preferences These options are accessed via the Select Utility Option menu Figure 5 below and include e SETUP Set the current date and time as well as the date and time formats e OUTPUT Specify where plate data should be sent to a printer a computer or to both Additional options include report format Column and or Matrix and whether or not to print standard curves e READ Enable or disable read time prompting for Plate ID Sample ID and READY 09 14AM 09 08 04 READ DEFINE REPORT UTIL SELECT UTILITY OPTION TESTS SETUP OUTPUT READ PROMPT FOR PLATE ID YES REPORT OUTPUT PRINT PRINT COMPUTER BOTH EDIT SETUP INFORMATION DATE TIME FILTER MORE Figure 5 Global configuration options available under UTIL Sample Count Specify whether or not to read in Rapid mode Note Refer to Using the Utility Options in Chapter 3 for more specific information on setting up the ELx800 ELx800 Operato
82. e and contact information of a person who may be contacted if questions arise 4 Close the box and tape it shut Write RMA and the RMA number in large clear letters on the outside of the shipping container and ship the instrument to the Bio Tek address provided in the Technical Support section of Chapter 1 ELx800 Operator s Manual 38 Chapter 2 Installation Bio Tek Instruments Inc Chapter 3 Operation This chapter includes instructions for operating the ELx800 and its software ELX8 00 Front Pali usciti leu eaaet tan Fats i a E rb aeu on den eames 40 OVOrFVIGW Sueciae toss erras beri durro eM e E d LC LPS ua ERE oO curl 42 Recommendations for Achieving Optimum Performance 42 System Startiupussscsvens ag quan cerae aco RR ASQ vedas van OR V A E ORO E 43 Mai MENU cc oo e idv ivdexzes os dx en e wists Rr de erede Qaa E A i 44 BI kde P EPI 46 Define Method Map Formula and Curve eeeeeeeeeee 49 Defining METHOD errare ana xa re yia X Rr n a nsa a aa 49 Defining MAP xcci isss cce coxa te ce VIS EE EX va idee Eua Publ uc tuus ut us ft 53 Defining FORMU LAG wast exe vx vcrb a X TER Rae o sa a Y EY es 69 Defining CURVE Vaer erano r ut cake duran EE ag wariita RAI PIENE 81 Reading a Microplate comes ene xau ecran dana cea CPUS au kB ER PER aa ERU E AD E 88 Selecting an Assay to RUN 1 cerebri erae nna i n e na ne 89 Printing RepOFLs sn itsen E ER XEYXEXRNX EX KEEN ERR ev ce
83. e edges Do not touch the glass portion of the filter Clean any filters that appear dirty with lens paper and isopropyl alcohol Once the filters are installed in the filter wheel place the wheel back on the pin in the base of the instrument making sure the filter wheel is sitting flat and that it meshes with the filter wheel drive gear Install the filter wheel cover with the four thumbscrews Note The cover can only be installed one way Place the top cover on the base and carefully turn the instrument upside down Reinstall the 4 screws with washers to hold the top cover in place Connect the power supply and cable to the rear of the instrument Important Store unused filters in a cool dry place away from direct sunlight The filters can be wrapped in a piece of lens paper to protect them from scratches and dust accumulation ELx800 Operator s Manual 28 Chapter 2 Installation Checking the Reader s Filter Table Setting After installing new filters ensure that the ELx800 s filter table the reader s software reference for filter locations matches the physical location of the filters To check or change the software filter table 1 Power up the reader At the Main Menu Screen press UTIL to display the SELECT UTILITY OPTION menu 2 From the SELECT UTILITY OPTION menu press SETUP The Edit Setup Information screen appears on the display SETUP INFORMATION T IME FILTER MOR E
84. e groove in the shipping block over the shaft and screw it down Figure 10 Shipping block Figure 10 Installing the shipping block ELx800 Operator s Manual 34 Chapter 2 Installation 8 Turn the instrument over and install the top cover that was removed in step 6 Figure 11 ae Top cover Figure 11 Reinstalling the top cover Bio Tek Instruments Inc Repackaging and Shipping the ELx800 35 9 Turn the instrument right side up and put the end caps on the unit Figure 12 Left end cap 26 x 32 2 Mil poly bag Figure 12 Reattaching the end caps 10 Place the unit with the end caps into the box Figure 13 ELx800 Operator s Manual 36 Chapter 2 Installation 11 Place the shelf into the notched out area in the end cap and the power supply into an 8 x 11 bubble bag Place the power supply on the shelf Figure 13 Figure 13 Placing the unit in the box Bio Tek Instruments Inc Repackaging and Shipping the ELx800 37 Preparing the Shipping Container 1 Obtain a Return Materials Authorization RMA number from Bio Tek s Technical Assistance Center through Bio Tek s Web site fax or e mail address listed in Chapter 1 2 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 3 Provide Bio Tek with the nam
85. e 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 ELx800 Operator s Manual 68 Chapter 3 Operation Number of Sample Replicates After the number of sample groups is entered the NUMBER OF SAMPLE REPLICATES entry screen is presented T ER NUMBER O F MP LIE 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 RE P 1 C0 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 Bio Tek Instruments Inc Define Method Map Formula and Curve 69 Defining FORMULA The ELx800 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 For
86. ear ELx800 Operator s Manual 86 Chapter 3 Operation 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 WN ACROSS This option ensures that all assays will be mapped in the same direction e Select DOWN or ACROSS The original mapping directions for the predefined assays are overridden 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 SELECT ASSAY NUMBER NAME H BS 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 Bio Tek Instruments Inc Define Method Map Formula and Curve 87 START MAPPING A T WELL LOCATION e Use the alpha and numeric keys to choose the well location to begin the assay Wells A01 through H01 are valid for ACROSS mapping A01 throu gh H12 are valid for DOWN e 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 e For example to include Assays 1 8 and 22 in the Pan
87. ed Warranty must be made in writing to Bio Tek Attention Service Department Purchaser must ship the Product to Bio Tek postage pre paid Bio Tek shall either repair or replace with new or like new at its option and without cost to the Purchaser any Product which in Bio Tek s sole judgment is defective by reason of defects in the materials or workmanship This Warranty is VOID if the Product has been damaged by accident or misuse or has been damaged by abuse or negligence in the operation or maintenance of the Product including without limitation unsafe operation operation by untrained personnel and failure to perform routine maintenance This Warranty is VOID if the Product has been repaired or altered by persons not authorized by Bio Tek or if the Product has had the serial number altered effaced or removed This Warranty is VOID if the Product has not been connected installed or adjusted strictly in accordance with written directions furnished by Bio Tek Batteries fuses light bulbs and other consumable items used in any of the Products are not covered by this Warranty Software utilized in conjunction with any of the Products is not covered by the terms of this Warranty but may be covered under a separate Bio Tek software warranty We will continue to stock parts for a minimum period of five 5 years after the manufacture of any equipment has been discontinued Parts shall include all materials charts instructions diagrams an
88. ed SERERE EXXNAS ERXXRAXEEYY 93 Editing Standard OUbllers issus depre enki shannon iter RY RYE 94 Printing ResllES irr rxY era ru serene sete YE EIS Sw E EQETA Sev d a EA tev 96 Using the Utility Options 1csixoekaue s odiis FUE Xa SR T ERE Sad veh luda vag e eu ETRd gas 97 Setting the Date and Time eia ree xk eia ea kw xa 97 Viewing Editing the Filter Table ccccccscseesseeessenseeeeseeeeees 98 Specifying Data Output and Reporting Options 99 Selecting Read Option si osevesso suns ve eaa ova eda S nc xS iD X i S T ese 101 40 Chapter 3 Operation ELx800 Front Panel READY 9 45AM 09 12 04 READ DEFINE REPORT UTIL CLEAR ENTER Previous Screen C Figure 14 Keyboard Bio Tek Instruments Inc ENTER Previous Screen CLEAR OOHE EE YW O N 9 Shift 7c ELx800 Front Panel 41 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 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 Pressing ENTER generally saves the current screen settings and adv
89. editable field and advances automatically The numeric range depends on the number of assays stored in the reader s memory The ELx800 has 55 open assays available the EL800 has only 10 2 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 below gt CLEAR Clears the reader s display gt MAIN MENU Returns the display to the Main Menu screen gt PREVIOUS SCREEN Returns the display to the previous screen gt ENTER Saves the current settings and advances to the next screen 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 Bio Tek Instruments Inc Define Method Map Formula and Curve
90. een 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 One cutoff formula may be defined per assay If Transformation Formulas are defined cutoffs are based on the transformed results Refer to Defining Formula on page 69 for the order in which formulas are processed 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 A greyzone around the cutoff value can be defined to indicate equivocal or indeterminate results Do not use the lt or gt mathematical symbols in a cutoff formula Tip Choose to print a Column Report to see the greyzone and cutoff values 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 0 of the cutoff value could be considered equivocal EQUIV EN TER GRE YZ2ONE 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
91. el assay DOWN mapping is selected for the Panel 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 Bio Tek Note The interpretation of Results reports for each assay in the Panel will print first and then the Sample results will print ELx800 Operator s Manual 88 Chapter 3 Operation 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 2 Note Before reading a plate ensure that the reporting options are set correctly under UTIL 3 OUTPUT To read a plate 1 Start at the Main Menu and
92. electronic noise were detected when blocking the light and increasing the gain to maximum Probable Causes e External signals may be penetrating the measurement chamber Ensure that the bottom and top shrouds as well as the filter wheel cover are correctly installed A faulty motherboard PN 7330400 7330410 or 7330415 or daughter board PN 7330404 or 7330414 Photodiode or power supply may be too close to the instrument Failure indicates excessive variation in the dark current background noise levels of the measurement circuit Reader failed offset 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 There may be an ambient light leak Ensure that the plate carrier door is properly closed Faulty motherboard PN 7330400 7330410 or 7330415 or daughter board PN 7330404 or 7330414 Photodiode or power supply may be too close to the instrument ELx800 Operator s Manual 146 Chapter 6 Troubleshooting and Error Codes General Errors Continued 0900 Read time dark value out of range e The dark current value taken during the current read is significantly different from the same reading taken during the power up self check Probable Cause e The measurement electronics background noise has changed since the last power up self check gt Ambient light may have increased si
93. ended Use a new microplate any fingerprints or scratches may cause variations in readings IMPORTANT Before running the liquid tests ensure that the reader is not running in Rapid Mode To check this select UTIL 99 READ and then cycle through the prompts until READ IN RAPID MODE is displayed Choose NO for an accurate result 1 Using freshly prepared stock solution A or B the concentrated solution prepare a 1 2 dilution using DI water one part stock one part DI water the resulting solution is a 1 2 dilution 2 Pipette 200 ul of the concentrated solution into Column 1 of the plate Pipette 200 ul of the diluted solution into Column 2 of the plate 4 Read the microplate five times at 405 nm using the Normal Read Mode Chapter 3 single wavelength no blanking 5 Print the five sets of raw data or export it to an Excel spreadsheet using KCjunior or KC4ATM If you are exporting the data to your own Excel spreadsheet perform the calculations described below and keep the spreadsheet for future tests Calculations 6 Calculate the mean and standard deviation for each well location in columns 1 and 2 for the five plates read Only one data set needs to be analyzed for each concentration The well that shows the most variation for each concentration is selected for data reduction 7 For each mean below 2 000 OD calculate the allowed deviation using the repeatability specification for a 96 well format o
94. entifier 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 0 Carrier X Axis 0 Reference 1 Carrier Y Axis 1 Measurement e A indicates a more 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 148 for more information Contact Bio Tek s Technical Assistance Center for further assistance if any of error codes are displayed Refer to Chapter 1 for contact information ELx800 Operator s Manual 142 Chapter 6 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 e User aborted from another serial interface A motor was unable to move to its home position as registered by feedback from an optical sensor Probable Causes Defective or broken optical sensor Flex cable is disconnected or damaged X axis movement is limited so that the optical
95. erformed following assay definition download Motor not homed successfully If the motor is not correctly homed the instrument sends an error and exits the function Probable Causes e Errors 0200 or 0300 have been ignored Refer to Probable Causes for these errors on pages 142 and 143 Fatal Errors Fatal Errors indicate conditions that require immediate attention If a fatal error is displayed contact Bio Tek s Technical Assistance Center for further instructions Refer to Chapter 1 for contact information Description and Probable Causes A100 Task control block not available A200 Read already in progress A300 Device not available Printer device not available A400 Failed code checksum on power up Code flash readback timed out A600 Quick flash configuration timed out Bio Tek Instruments Inc Appendix A Controlling the Reader With KCjunior or KC4 The ELx800 can be controlled either from Bio Tek s KCjunior or KC4 PC Software This Appendix provides instructions for programming the computer to control the reader OVERVIEW ucssteden uou tes vasa su ola Raa dd Ves bee awa OR EPOD mae miM went 150 Controlling the Reader With KCjunior s sssssssssssserrrrrrrrrrsrrreress 151 Problems i ud urcesrc turo oO nad a rbd ba VIR ACRI au RES Feed aE 152 Getting Started With KCjunior esee nenne 152 Controlling the Reader With KC4 sssssssssssssseeeen nnn
96. erwrite the first Plate ID stored in memory Bio Tek Instruments Inc Reading a Microplate 91 e Use up to 10 alphanumeric characters See page 40 for instructions on using the keypad e 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 RE P 1 G01 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 ELx800 Operator s Manual 92 Chapter 3 Operation 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 R E AD 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
97. ess ENTER to save the value and continue 3 When prompted insert the Test Plate into the plate carrier and press the READ Key to begin the test The Calibration Plate Analysis Report Figure 19 will be sent to a printer when the test is run Bio Tek Instruments Inc Qualification Procedures 115 To run the Absorbance Plate Test in KC4 1 Open the System menu and select Diagnostics Run Universal Plate Test 2 Select the Universal Plate and then click Run Test To run the Absorbance Plate Test in KCjunior 1 Open the Utilities menu and select Diagnostics Universal Plate Test 2 Select the Universal Plate ID and then click Run Test Test Failures If any of the test parameters report as FAIL confirm that the standard values on the test plate data sheet match the values on the printout If not correct and retest If the test still fails contact Bio Tek 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 ELx800 Operator s Manual 116 Chapter 4 Performance Verification and IQ OQ PQ Procedures Liquid Testing Conducting Liquid Tests confirms the ELx800 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 opt
98. f x 0 596 0 005 OD ELx800 Operator s Manual 120 Chapter 4 Performance Verification and IQ OQ PQ Procedures 8 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 0 5 1 951 0 005 0 0098 which when added to the 0 005 0 0098 0 005 0 0148 OD which is the allowable deviation Since the standard deviation is less than this value the reader meets the test criteria Repeatability Specification For comparison in this test the following repeatability specification is applied using the Normal Read Mode and a 96 well microplate 0 5 0 005 OD from 0 000 to 2 000 OD 405 nm Bio Tek Instruments Inc Liquid Testing 121 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
99. f 41 samples to be tested in duplicate or 89 in single with a standard curve and positive and negative controls Background Anticardiolipin antibodies are found in a wide range of conditions either transiently in some infectious diseases or more persistently in autoimmune diseases such as systemic lupus erythematosus SLE and antiphospholipid syndrome APS Anticardiolipin antibodies have also been associated with a range of clinical conditions including fetal loss endocarditis stroke heart attack and autoimmune haemolytic Principle of the Assay Microwells are precoated with cardiolipin and cofactor Standards controls and patient samples are added to the wells and autoantibodies recognizing cardiolipin bind during the first incubation After washing the wells to remove all unbound proteins conjugate is added The conjugate binds to the captured human antibody and the excess unbound conjugate is removed by a further wash step Substrate is added that causes a blue reaction thereby exposing the bound conjugate and producing an intensity proportional to the concentration of autoantibody in the sample Phosphoric acid is added to each well to stop the reaction This produces a yellow end point color which is read at 450 nm ELx800 Operator s Manual 180 Appendix E Instructions for Programming a New Assay Materials Materials Supplied Instruction Leaflet Giving full assay details QC Certificate Indicating the expe
100. filter table see Chapter 2 Filter signal out of range The filter 1 5 has a signal that is out of range Probable Causes e This indicates a light level problem with one or more filters e A UV filter may have been installed in a non UV instrument e A blank filter may be incorrectly installed in a filter position Verify that filters in the filter wheel match settings in the filter table see Chapter 2 Configuration data missing Required reader configuration data was missing at the beginning of the self test Possible Causes e Necessary configuration data was not downloaded or was downloaded incorrectly ELx800 Operator s Manual 148 Chapter 6 Troubleshooting and Error Codes General Errors Continued Failed configuration checksum test The stored checksum value from the configuration data does not match the actual checksum of the current configuration data Probable Cause e The configuration data has changed and the checksum stored is no longer valid The error is produced when outdated versions of assay definition software are used to create an assay configuration file This file is incompatible with the operation code within the instrument s memory Recreate the assay definition using the correct version of assay definition software and re download the software Autocalibration data missing No Autocal data exists for the read location Probable Cause e Autocalibration was not p
101. for the next control Number of Control Replicates The NUMBER OF CONTROL REPLICATES entry screen is displayed if the number of control groups is greater than 0 EN TER NUMBER REPLICATES 0 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 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 Bio Tek Instruments Inc Define Method Map Formula and Curve 67 Location of Controls Use this option to enter the location of controls in the assay CONTROL 1 LOCATION TY PE PC RE P 1 A0 2 The displayed location field can only be edited if manual mapping was selected see page 54 This screen is not accessible in auto map 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 READ PROMPT FOR SAMPLE COUNT is set to YES See Selecting Read Options on page 101 for more information e Use the numeric keys to enter the number of sample groups on the plate e The rang
102. formation and Formula calculations for more complex mathematical operations including validations e Up to 75 assays can be programmed into memory and recalled instantly e Up to 10 results can be stored in memory ELx800 Operator s Manual 4 Chapter 1 Introduction Package Contents The contents of the ELx800 package include Microplate Reader Power Cord part number varies according to country of use Power Supply Filter wheel with 4 standard filters 405 nm 450 nm 490 nm 630 nm and one blank filter The UV model includes a 340 nm filter Operator s Manual PN 7331000 and Registration Card Printer Cable PN 71072 Dust Cover PN 7332040 RS 232 Serial Cable PN 75053 Unpacking instructions PN 7271003 packing instructions PN 7271012 Shipping document kit PN 94075 including Warranty Registration Card Certificate of Compliance and Calibration and FDA Certification Form Set of shipping materials PN 7273000 Declaration of Conformity PN 7271004 Sample UV transparent plates and literature Bio Tek Instruments Inc Optional Accessories 5 Optional Accessories e Patented Bio Cell Quartz vessel to obtain 1 cm fixed pathlength absorbance measurements PN 7272051 e Bio Cell adapter plate assembly can contain up to 8 Bio Cells PN 7270512 e Lamp for ELx800NB ELx800RNB ELx800UV ELx800RUV PN 7330516 e Lamp for ELx800 and ELx800R PN 7330513 e ELx800 filters PN 7334 plus
103. g the decontamination process must be familiar with the basic setup and operation of the instrument Bio Tek 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 Bio Tek nor the 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 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 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 ELx800 Operator s Manual 136 Chapter 5 Maintenance and Decontamination Tools and Supplies e Sodium hypochlorite NaClO 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 Bioh
104. hanged printer compatible description Changed the number of samples necessary if there aren t controls standards or blanks defined Corrected the number of stored plates to 8 Corrected the positions for re use of standard curves Removed the M command from Appendix B Computer Control Added Liquid Test 3 to test 340 nm filters Updated screens in Chapter 3 Updated chart in Appendix F Preface Updated contact information in Notices page iii Added Document Conventions page v Corrected errors in Safety Symbol text page ix Removed screwdriver from Package Contents and added Bio Tek QC Check Solutions for 405 nm to Optional Accessories list page 1 7 Removed About This Manual section page 1 5 Updated contact information in Technical Support Chapter 1 Modified appearance of display screens throughout Revised lamp alignment section in Chapter 2 xiv Preface Revision History Continued L 5 03 Chapter 4 Cont Changed title to Performance Verification and IO PO OO Tests Added IQ PO OQ test procedure information Clarified procedures for liquid tests Replaced outdated Appendix E with instructions for using KCjunior to read 384 well microplates Added new Appendix G with two examples of assay kit instructions and directions for programming an assay Made editorial and formatting changes throughout M 9 03 Preface Updated Warnings section pages vii and viii Updated Electromagnetic Com
105. haude Warnen hei e Oberfl che 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 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 Bio Tek Instruments Inc Warranty xxiii Warranty This Warranty is limited and applies only to new products except for computer based software which is covered under a separate Warranty Policy manufactured by Bio Tek Instruments Inc Bio Tek Bio Tek makes no warranty whatsoever regarding the condition of used products Bio Tek warrants the instrument hereinafter collectively referred to as Products or Product for a period of one 1 year from the original purchase date against defective materials or workmanship This Warranty is limited to the original purchaser the Purchaser and cannot be assigned or transferred All claims under this Limit
106. ics 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 e Liquid Test 1 confirms repeatability and consistency of the reader when a solution is used in the microplate If these tests pass then the lens placement and system cleanliness are proven e 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 121 e Liquid Test 3 is an optional test and can only be performed with the UV model It is offered for those sites that must have proof of linearity at wavelengths lower than those attainable with the Absorbance Test Plate This test is optional since the ELx800 has good front end linearity throughout the specified wavelength range see page 125 e Bio Tek 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 3 page 121 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 e Alternatively any solution that gives a stable color will suffice This includes subs
107. ied 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 STANDARD S Y E Y ES 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 Bio Tek Instruments Inc Define Method Map Formula and Curve 63 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 R E P 1 A0 1 e Use the numeric and alpha keys and the decimal point key to enter standard concentration values The range is 0 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
108. internal self checks e Excessive ambient light Bright sunlight or strong incandescent light can reduce the linear performance range of the instrument e Dust Optical density readings may be affected by extraneous particles such as dust in the microplate wells A clean work area is necessary to ensure accurate readings Electrical Connections Caution Power Supply Only use the specified power supply to ensure proper operation of the unit The ELx800 has a universal 24 VDC 2 Amp power supply that functions from 100 to 240 V 10 0 50 to 60 Hz without external switching Never use a two prong plug adapter to connect primary power to the ELx800 power supply Use of a two prong adapter disconnects the utility ground creating a severe shock hazard Always connect the supply power cord directly to a three prong receptacle with a functional ground 1 Connect the power cord to the external power supply 2 Plug the rounded end of the power supply line cord into the power supply jack on the rear of the instrument 3 Tighten the knurled nut on the power supply outlet to ensure that the plug 4 Plug the does not pull out 3 prong end of the power cord into an appropriate power receptacle Warning Power Rating The ELx800 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
109. istilled water and wipe all surfaces of the instrument that have been cleaned with the bleach solution or alcohol 7 Usea clean dry cloth to dry all wet surfaces 8 Discard the used gloves and cloths using a Biohazard trash bag and an approved Biohazard container ELx800 Operator s Manual 138 Chapter 5 Maintenance and Decontamination Bio Tek Instruments Inc Chapter 6 Troubleshooting and Error Codes This appendix describes potential error codes that may be displayed on the ELx800 and suggests possible solutions for these problems OVERVIEW itisid ri aniu sence E a cR MR a E LIV Ma tad AC Dln 140 System Test DeScrIDLIOT assis xxiii ca e RE ROGER ER OCDE EN Ra Ra seen Dac 140 Glossary of Ters asturias ne dO koe t ot ic Ea A de 140 EFPOF CCOCGS neueste ius dla RR LU ERE OR AAE EREET ONEA NEE inde RD E RaT 141 General cago AC EET 142 E talEELOES 33 2 acc dicere eec ex expo kA E in baa rave dadas 148 140 Chapter 6 Troubleshooting and Error Codes Overview If an error is displayed 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 Bio Tek 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 five set wavelengths which confirm adequate ligh
110. k Instruments Inc Define Method Map Formula and Curve 59 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 column 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 m
111. k Instruments Inc Define Method Map Formula and Curve 85 Panel Assays A Panel assay is a collection of up to 8 assays to be run on one plate The most common reason to use a Panel assay is for confirmatory tests based on a screening test in clinical applications Only one panel can be defined on the reader at any time The assays specified within the Panel must be predefined in any of the assay positions 1 55 The assays specified within the Panel must all use the Endpoint read method The assays specified within the Panel must all read at the same wavelength s Any curve fit type formulas or standard concentrations previously defined for each assay will be used when the assay is selected for a Panel Panel assays cannot reuse standard curves 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 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 The default name is PANEL Use the alpha and numeric keys to update the assay name if desired Press ENTER to continue The NUMBER OF ASSAYS screen will app
112. k representative or Bio Tek Technical Support 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 filters on the test plate to ensure that there is no debris that may have shifted between readings and caused changes gt Check the microplate carrier to ensure that it is clear of debris gt If these remedies are ineffective call your Bio Tek representative or Bio Tek Technical Support ELx800 Operator s Manual 112 Chapter 4 Performance Verification and IQ OQ PQ Procedures Requirements To run the Absorbance Plate Test you need Bio Tek s Absorbance 7 Filter Test Plate Part Number 7260522 with its accompanying Data Sheet shown in Figure 18 Figure 19 shows a sample Test Plate Analysis Report This test plate can be used for testing the reproducibility linearity and alignment of your Bio Tek autoreader The following calibration data has been recorded by a N I S T traceable spectrophotometer WAVELENGTH nm wen 405mm 450nm 49onm ssonm 620mm esonm f Boni zoom 91r ono ors et o1 ore os et EH ERG q
113. k the Test Communications button to attempt to establish communications with the reader using the currently defined communication parameters If a Serial Write Error dialog box is displayed an incorrect Com Port may have been selected Select a different port and then repeat this step If the test passes click OK to save the settings and close the dialog box If the test fails follow the directions provided by KCjunior then click Test Communications again To learn more about KCjunior and how to define assays and read plates refer to the KCjunior User s Manual ELx800 Operator s Manual 152 Appendix A Controlling the Reader With KCjunior or KC4 Problems If KCjunior fails to communicate with the reader and displays a serial communications error check the cable plug in location to make sure it matches the setup choices and is not a Null cable If this is suspected add another Null and try again Getting Started With KCjunior The following instructions briefly describe how to read a plate using KCjunior Refer to KCjunior s Help system and User s Guide early and often to learn how to create protocols assign well identifiers read plates print reports and more To read a plate using KCjunior 1 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
114. le 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 Bio Tek Instruments Inc Using the Utility Options 97 Using the Utility Options 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 EDIT SETUP INFORMATION DATE T IME FILTER MO 2 Select DATE The DATE entry screen will appear 01 23 04 DDMMYY 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 To change the current time and or the time format select TIME from the EDIT SETUP INFORMATION menu The TIME entry screen will appear gt Enter the time using the numeric keys on the keypad The cursor is positioned under the first editable field and
115. 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 Warranty Failure to follow preventive maintenance protocols may void the warranty 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 Bio Tek Instruments Inc Hazards and Precautions xix C Based on the testing described below and information contained herein this instrument bears the CE mark Directive 89 336 EEC Electromagnetic Compatibility Emissions CLASS A EC 61326 1 2002 EN 55022 2000 Class A CISPR 16 1 1993 CISPR 16 2 1999 Immunity IEC 61326 1 2000 EN 61000 4 2 Electrostatic Discharge EN 61000 4 3 Radiated EM Fields EN 61000 4 4 Electrical Fast Transient Burst EN 61000 4 5 Surge Immunity EN 61000 4 6 Conducted Disturbances EN 61000 4 11 Voltage Dips Short Interruptions and Variations Directive 73 23 EEC Low Voltage EN 61010 1 2001 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
116. lt can be printed by choosing Report from the Main Menu screen 164 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 Figure 31 Samples with calls on Matrix Report Bio Tek Instruments Inc 165 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 A B 0 R B A N C E CONCENTRATION Curve y Ax 8 Coefficients A 0 035 Bz 132 R Sqr 1 0000 Figure 32 Curve Fit Report ELx800 Operator s Manual 166 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 GREYZONE 0 20 0 200 CUTOFF 1 1 000 SMP SMP lt CUTOFF CUTOFF GREYZONE AND SMP z CUTOFF CUTOFF GREYZONE EQUIV SMP SMP gt CUTOFF CUTOFFSGREYZONE NEG SMP SMP CUTOFF CUTOFF GREYZONE POS PC PC gt 01 Well 1D CalcoD Call Pred Conc RSLT Std Dev MEI PC 0 109 VAL B01 PC 0 119 VAL 0 114 C01 SMPOO01 0 290 POS DOI SMP0002 0 301 POS E01 SMP0003 2 205 NEG F0i MP0004 2 811 NEG G0 SMPO005 3 499 HO1 SMPO006 3 225 A02 SMP0007 0 085 B02 MPO008 1 054 C02 SMPO009 1 092 D02 SMP0010 2 075 E02 SMP0011
117. main menu System Readers 3 The Reader Selection dialog will appear Select the ELx800 reader and define the communications parameters Click on Close to return to the main menu Reader Selection X Current Reader ELx800 on COM2 Close Available Readers Help ELx808 on COM2 ELx808 on COM2 B FL600 on COM2 FL 800 on COM2 FLx800 1 on COM2 Powerwave 200 on COM2 PowerwW ave 200 1 on COM2 Figure 27 Reader Selection dialog ELx800 Operator s Manual 160 Appendix B Using 384 Well Geometry 4 Select Data New Plate itekc4 Data Edit Protocol Display System New Plate CtriN Open Ctr Clase Gaye Gres Save As Read Plate crite Sign Export Append To Kinetic File Zi z Bepo options Biogrsph Control Printer Ctrl P Print Preview Print Setup Plate Description Samples Audit Trail Exit ELx800 on COM2 System Administrator NUM Figure 28 Data New Plate dialog 5 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 New Data File x Select a Protocol New Test NewProtocol Test Protocol Qualit Protocol Path Cancel Browse Hep Figure 29 New Data File dialog Bio Tek Instruments Inc 6 10 11 12 13 kc4 161 Select Protocol
118. 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 ELx800 Operator s Manual 72 Chapter 3 Operation MATH The following mathematical symbols can be used in formulas Equal to Greater than Greater than or equal to Addition ey l Subtraction 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 Less than or equal to FUNCTION The following functions can be used in formulas LOG10 Log Base 10 ALOG ALOGIO 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 ALOG10 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 Bio Tek Instruments Inc Define Method Map Formula and Curve 73 Validation
119. ment may be used for clinical research and development or other nonclinical purposes If there is no such label then the instrument may only be used for research and development or for other nonclinical purposes Quality Control It is considered good laboratory practice to run laboratory samples according to instructions and specific recommendations included in the assay package insert for the test to be conducted Failure to conduct Quality Control checks could result in erroneous test data Repackaging and Shipping or repair contact Bio Tek 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 Bio Tek for replacement packing If you need to ship the instrument to Bio Tek for service ELx800 Operator s Manual xvi Preface Warnings General e Operate the instrument on a flat surface away from excessive humidity Readers e Excessive Ambient Light Bright sunlight or strong incandescent light can reduce the linear performance range of the instrument e Dust Measurement values may be affected by extraneous particles such as dust in the microplate wells A clean work area is necessary to ensure accurate readings e When operated in a safe environment according to the instructions in this document there are no known ha
120. move the optics arm cover Loosen the hex screws that hold the lamp in place Do not remove the screws Slide the bulb out backward and detach it from the connector at the back of the optics arm Insert the new bulb into the bulb retention springs Plug in and power up the unit The lamp should light although an error message will appear on the instrument s display because the lamp is not aligned Press any key to return to the Main Menu To align the bulb 1 From the Main Menu press UTIL to access the SELECT UTILITY OPTION screen At the SELECT UTILITY OPTION screen press SETUP gt READ gt MORE until BULB ALIGN is displayed The carrier will move to the bulb alignment position Bio Tek Instruments Inc Replacing the Bulb 133 Bulb assembly Optics arm Filter wheel cover Lamp retention screws p __ Lead wires 1 aperture Optics arm cover a Optics arm _ A Y Lamp retention screws Figure 20 Lamp assembly ELx800 Operator s Manual 134 Chapter 5 Maintenance and Decontamination To align bulbs on a standard model reader AN A Warning The lamp will be hot immediately after the reader has been shut down Allow the lamp to cool down for at least 10 minutes before attempting to replace the bulb Use the lead wires to position the bulb to avoid damaging the glass covering the bulb with fingertips Push the bulb forward until it stops Rotate and or swing the
121. mples defined for this assay the map will be automatically regenerated without the standards beginning in well xxx where xxx was chosen as the starting well in the map usually well A01 If manual mapping was used to map the plate the map is not regenerated the reader will not produce results for the well positions that originally were standards Auto mapping is recommended if the standard s curves will be routinely reused 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 locations on the plate that are undefined The maximum number of control groups is 8 ELx800 Operator s Manual 66 Chapter 3 Operation Control Type This option allows you to enter the type of control used in the assay Any previously defined Control Type will be displayed on the screen 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
122. mula definition is not available in the EL800 version To define formulas 1 Start at the Main Menu and select DEFINE 2 Select the assay then press ENTER to display the DEFINE options screen FORMULA CURVE 3 Select FORMULA The SELECT FORMULA TYPE screen will appear described on the next page 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 Blank Validation 0 1 Control Validation 0 4 Assay Validation 0 4 Transformations 0 1 Cutoff Formulas 0 1 Curve Fit Analysis if a curve fit method is defined To capture and manipulate the raw data using 384 well microplates with the ELx800NB you must use KCjunior or KC4 KCjunior and KC4 are PC based software programs you can use to set up your assay communicate with the ELx800 to run the assay and then manipulate the raw data that is automatically retrieved from the reader See Refer to Appendix B for additional information ELx800 Operator s Manual 70 Chapter 3 Operation Formula Type The ELx800 supports three types of formulas as well as the ability to define variables for use within Transformation formulas Note GENERAL formulas are not used 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
123. n the assay If the assay has a closed variable the screen for this variable is omitted FORMULA CURVE 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 GENERATION 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 manual map Bio Tek Instruments Inc Define Method Map Formula and Curve 55 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 e Select DOWN to map down the column e Select ACROSS to map across the row Press ENTER to save the selection and continue Replication Direction This option allows you to
124. nce Center 800 242 4685 Bio Tek Instruments Inc Technical Support 11 Electronic Communication Electronic communication is available via the following E Mail tac biotek com Internet Site www biotek com Facsimile Support You may send a fax with your questions or requests for help 24 hours a day to the following number Technical Assistance Center 802 655 3399 Written Communication If you prefer you may write a letter with your comments and send it to Bio Tek Instruments Inc Technical Assistance Center P O Box 998 Highland Park Winooski Vermont 05404 0998 USA Shipping Address Ship instruments that need repair or service to Bio Tek at the following address Bio Tek Instruments Inc Technical Assistance Center 100 Tigan Street Highland Park Winooski Vermont 05404 USA ELx800 Operator s Manual 12 Chapter 1 Introduction Bio Tek Instruments Inc Chapter 2 Installation This chapter includes instructions for unpacking and setting up the ELx800 and instructions for connecting printers and or serial devices Unpacking and Inspecting the ELX800 cccceceeeeeeeeeeeeeeeeeeees Unpacking the Instrument and Its Accessories Setting Up the ELX8QO wes cred evr vate eda ev XR X dod ea Va DE RR tas Operating Environment ssseessssssssen e nnne n nnn Electrical Connections oss fcc utc eor RETENEN c eese Power Up and System Test
125. nce power up 0AO01 Read time Air Blank out of range ano The blank full signal reading taken during the current read has changed significantly from the same reading taken during the power up self check Probable Cause e Lamp is unstable Reboot system and or install new lamp Turn unit OFF when not in use to conserve lamp life OBOO Invalid assay An assay number that is not programmed was selected 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 Bio Tek Instruments Inc Error Codes 147 General Errors Continued Errors ODOO through 1300 indicate that the reader has failed one or more of a number of self tests Contact Bio Tek s Technical Assistance Center for instructions Refer to Chapter 1 for contact information Calibration checksum error Reader failed calibration checksum test Probable Cause e The stored checksum value for the calibration data does not match the actual checksum 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 e Assay wavelength does not match the filter table gt Run system test Verify that wavelengths in the filter wheel match settings in the
126. nd Parity settings cannot be changed Bio Tek Instruments Inc Setting Up the Serial Port for Communications With Other Devices 25 To change the baud rate from the instrument keypad 1 Atthe Main Menu press UTIL 12 45 PM 01 23 DEFINE REPORT U 2 Atthe SELECT UTILITY OPTION screen press SETUP ELECT UTILITY OPTION ESTS SETUP OUTPUT R E 3 Atthe EDIT SETUP INFORMATION screen press MORE and then RS232 to continue 4 The SELECT BAUD RATE screen will appear showing the currently defined Baud Rate ELECT BAUD RATE 2 0 0 2400 9600 Select the desired baud rate Select VIEW if you wish to see the reader s other communication settings RS 232 SETTING 2 STOP BITS To change the baud rate or other communications settings in KC4 or KCjunior refer to their respective user guides or to Appendix A Computer Control ELx800 Operator s Manual 26 Chapter 2 Installation Installing Additional Filters Installed in the internal five position filter wheel are the filters that come standard with the ELx800 standard models have 405 450 490 630 nm filters the UV model s filter set is 405 450 490 630 and 340 nm Important Keep track of all filter locations The physical location of the filters must match the filter locations mapped in the reader s software filter table The filter wheel must have no empty locations all locations must be filled with either a
127. ndows can control the ELx800 in 6 12 24 48 96 and 384 well formats as well as in 60 72 and 96 well Terasaki plate formats 1 Start KCjunior 2 At KCjunior s main menu choose Setup Reader 1 KCjunior Results Protocols Utilities Setup CurrentReader View Help Reader 1 mmm Reader 2 Configuration Figure 21 KCjunior main menu Setup Reader 1 dialog 3 The Reader Setup 1 dialog will appear Select ELx800 EL800 at Reader Type and define the communication parameters Click OK to return to the main menu screen Reader Setup 1 X Reader Type ELx800 EL800 IM f Cancel Communications Settings Help come Com Port Baud Rate 9600 Get Comm Defaults Data Bits e E Test Communications j asec y j Parity None z Get Basecode Version Stop Bits 2 EOT Character 26 0014 Figure 22 Reader Setup 1 dialog Bio Tek Instruments Inc KCjunior 157 4 Click Read Plate to enter the Results ID and Plate Description Read Plate Dialog Feader Read Plate JEL 800 E L800 Simulation Data Protocol Name Read From File SAM PLE Cytotoxicity Il Cancel Results ID Help Plate Description Figure 23 Read Plate dialog 5 Choose New Protocol to define the protocol Protocol Definition o xl General Information Read Method Template Curve Cutoffs Reports Protocol Name SAMPLE Cytotoxicity Ill
128. 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 modifying software parameters and establishing reading or wash methods using the instrument s onboard software Failure to conduct quality control checks could result in erroneous test data Warning Reader Data Reduction Protocol For readers with onboard assay software the 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 For readers 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 ELx800 Operator s Manual xviii Preface Precautions The following precautions are provided to help avoid damage to the instrument should service the instrument Only qualified technical personnel should perform troubleshooting and service procedures on internal components Caution Service Only Bio Tek authorized service personnel Caution Environmental Conditions Do not expose the system to temperature extremes For proper operation ambient temperatures should remain between 15 40 C Performance may be adversely affected if temperatures fluctuate above or below this range Storage temperature
129. notes Warning A Warning indicates the potential for bodily harm and tells you how to avoid the problem Caution A Caution indicates potential damage to the instrument and tells you how to avoid the problem DEFINE Text in COURIER font represents 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 Bio Tek Instruments Inc Revision History xiii Revision History Revision A muUo w ri ELx800 Operator s Manual Date 3 95 7 95 9 95 10 95 11 96 2 97 03 98 09 98 2 99 9 99 1 00 5 08 Changes First Release Generic Add Errata sheet Incorporate Errata cal plate PN cover text to UV Added Panel Assay info Added TVar parameter Added Appendix D to include all reports Added Notes including info on 384 well format Updated Reuse of Standard Curves and Panel sections Added Appendix E KcJr for 384 well format Added Liquid Test 1 and 2 to Chapter 4 Performance Verification Added Appendix F Comparison to show the differences between the ELx800 and the EL800 Added printer information Changed plate type information to include 384 well and 72 96 well Terasaki plate formats Added reference to Quick Read on display Modified Appendix B Computer Control Updated EIx800 and EL800 model comparison table in Appendix F Incorporated Manual Updates Changed European addresses C
130. nu Press RESULT select the assay and then press ENTER The EDIT STD OUTLIERS YES NO prompt will appear See Editing Standard Outliers on page 94 for further instructions ELx800 Operator s Manual 84 Chapter 3 Operation X Y Axis Type Note LIN LIN axes are recommended when using the Cubic Spline fit After the curve fit type is selected select the X Y Axis Type AXIS T YP E LIN LIN LOG LOG LOG LIN 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 EX TRAPOLAT E 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 44 425 If extrapolation is chosen for the Point to Point curve fit unknown concentrations will be extrapolated linearly from the nearest segment of the curve If the plot includes both increasing and decreasing segments the curve printout will be labeled Ambiguous The resulting values which actually are extrapolated may not be indicated as such All calculated results for an Ambiguous curve should be considered unreliable Bio Te
131. ot 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 readers support standard flat U bottom and V bottom microplates optimum performance is achieved with optically clear flat bottomed wells Non uniformity in the optical density of the well bottoms can cause loss of accuracy especially with U and V bottom polyvinyl microplates Check for this by reading an empty microplate Dual wavelength readings can eliminate this problem or bring the variation in density readings to within acceptable limits for most measurements 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 ina 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 limits Use Tween 20 if possible or some other wetting agent to normalize the meniscus Some
132. ove the instrument from the plastic bag Unpacking instructions Left end cap Unit inside 26 x 32 2 Mil poly bag Right end cap Figure 2 Removing the foam end caps Bio Tek Instruments Inc Unpacking and Inspecting the ELx800 17 4 Carefully turn the ELx800 upside down on a level surface Use the slotted screwdriver to remove the four screws from the top cover mounting Figure 3 and shipping straps from the bottom of the instrument Set aside these screws Shipping straps hoN Top cover Figure 3 Removing the top cover mounting screws ELx800 Operator s Manual 18 Chapter 2 Installation 6 Turn the instrument right side up 7 Remove the shipping block Figure 4 Figure 4 Removing the shipping block 8 Replace the top cover Turn the instrument upside down and replace the mounting screws 9 Remove the shipping straps from the top of the carrier Save these for repackaging shipping Bio Tek Instruments Inc Setting Up the ELx800 19 Setting Up the ELx800 Operating Environment The ELx800 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 e Excessive humidity Condensation directly on the sensitive electronic circuitry can cause the instrument to fail
133. patibility section page x Added Consult instructions for use and In vitro diagnostic medical device safety symbols page xii Expanded Intended Use Statement page xiii Added Absorbance Test Plate to Optional Accessories list Chapter 1 page 1 8 Changed callout in Figure 2 2 from Place unit in bag to Unit in plastic bag Chapter 2 page 2 2 Added KC4 startup information to Appendix E Removed references to ELx800UV and EL800UV from Appendix F Standardized the presentation of significant digits throughout Changed Abs to OD throughout N 10 04 Restructured manual according to new template Removed references to General Formula throughout Updated Chapter 4 Performance Verification and IO OQ PO Procedures Added new Chapter 5 Maintenance and Decontamination Added new Chapter 6 Troubleshooting and Error Codes Removed Computer Control portion of Appendix A renamed Appendix to Controlling the Reader With KCjunior or KC4 Added new Appendix B Using 384 Well Geometry Updated sample reports in Appendix C Report Format formerly Appendix D Removed previous Appendix A Decontamination Removed previous Appendix C Error Codes Added new Appendix E Instructions for Programming a New Assay Bio Tek Instruments Inc Intended Use Statement xv Intended Use Statement The intended use of this instrument is dependent on the instrument s rear panel label If there is an IVD label then the instru
134. ps from the home position and back Probable Causes e The belt has slipped because of incorrect tension a loose motor pulley or a loose belt clamp e Defective motor drive Carrier Y 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 belt has slipped because of incorrect tension a loose motor pulley or a loose belt clamp e Defective motor drive Filter wheel failed positional verify lamp failed Probable Causes e Optical trigger flag has moved or is loose e Filter wheel is binding against the motor gear e Defective motor drive e Bearings caused motors to fail Filter not installed Probable Causes e Empty filter location on the filter wheel all filter locations must have either a filter or a filter blank Bio Tek PN 2872086 Entire filter wheel is not installed Bio Tek Instruments Inc Error Codes 145 General Errors Continued oso Ferat Gain outofrenge OOOO 0601 Filer 2 Gainoutofrange ooo 0603 Filter 3 Gain outofrange o ooo Dee These errors indicate that the gain for the filter in question is out of the range necessary to ensure performance to specifications Probable Causes e Defective interference filter e Defective lamp providing reduced signal causing the error e Misaligned optics Reader failed noise test Significant variations in background
135. r P N 7330531 is available from Bio Tek if Terasaki plates are to be used with the NB reader ELx800 Operator s Manual 52 Chapter 3 Operation Press MORE to cycle through the available options 6 6 well 2 x 3 96 96 well 8 x 12 12 12 well 3 x 4 96H 96 well Hellma Quartz 8 x 12 24 24 well 4 x 6 96M 96 well Metric 8 x 12 9 mm well 48 48 well 6 x 8 spacing Bio Tek Instruments Inc Define Method Map Formula and Curve 53 Defining MAP The MAP GENERATION 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 The NB reader using 384 well geometry does not have access to map formula or curve options The 384 well plate type is preset for the map to have 384 samples This does not include any blanking controls or standards Selecting 6 12 24 48 or 96 well geometries enables data reduction capabilities ELx800 Operator s Manual 54 Chapter 3 Operation MAP screens appear in the order that they were defined i
136. r just to the sample wells SMP COP E VARIABLE D ELx800 Operator s Manual 76 Chapter 3 Operation 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 transformation formula The TVAR will be available as a MAP option when building the transformation formula Example The assay plate map has 2 blanks 1 control well in duplicate CTL1 1 negative control well in triplicate NC and 5 standards in duplicate STD1 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 Bio Tek Instr
137. r s Manual 22 Chapter 2 Installation Connecting a Printer to the ELx800 Connect the printer to the ELx800 only if you are running the instrument in standalone mode If you are using Bio Tek s KCjunior or KC4 software skip this step and go to page 24 Setting Up the Serial Port for Communications The ELx800 has a parallel port LPT1 to allow connection to Epson compatible printers or HP Deskjet printers See Specifying Data Output and Reporting Options in Chapter 3 for more information The port is illustrated in Figure 6 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 ELx800 To attach a printer to the ELx800 Turn the reader off Place the printer in a location adjacent to the ELx800 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 ELx800 Tighten the securing screws on both ends of the cable pe UL ae cum he 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 Note For the latest list of compatible printers consult the Bio Tek Web site www biotek com or call Bio Tek Instruments Technical Assistance Center refer to Chapter 1 for contact information Bio Tek Instruments Inc Setting Up the ELx800
138. red 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 Precision balance with readability to 0 01 g e Buffer solution A or B Solution A 10x Concentrate Phosphate Buffered Saline PBS Required Materials e Deionized water e Ingredients shown in Table 4 e p 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 ELx800 Operator s Manual 126 Chapter 4 Performance Verification and IQ OQ PQ Procedures 1 Prepare the stock buffer solution using the ingredients in Table 4 Table 4 PBS 10X Concentrate Solution NaCl KCI qp sog RO wm 2 Mix5 ml of the concentrated PBS solution with 45 ml of DI water 3 Add 10 mg of the B NADH powder and mix thoroughly This is the 10x Concentrate PBS Solution Solution B Sigma PBS Required Materials e Deionized water e Tween 20 polyoxyethylene 20 sorbitan monolaurate e Sigma P 3563 packets e B NADH Powder B 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 1 Prepare a PBS solution
139. riteria may lock up during the Calculating Results phase of data reduction see Beginning the Plate Read on page 92 This can be corrected by modifying assay criteria as follows e Specify at least one 01 sample when mapping the plate and or e Change PROMPT SAMPLE COUNT to YES e Set CURVE FIT to NONE if you do not have a standard curve as part of the assay 1 From the Main Menu press DEFINE gt MAP and continue to press ENTER until ENTER NUMBER OF SAMPLES is displayed Enter at least one sample The software will lock up if zero 00 samples are selected ENTER NUMBER O F SAMPLES 2 From the Main Menu press UTIL 9 READ and continue to press ENTER until PROMPT SAMPLE COUNT is displayed Select YES PROMPT SAMPLE COUNT Y N O 3 From the Main Menu press DEFINE SELECT ASSAY NUMBER gt ENTER At the DEFINE menu press CURVE to display CURVE FIT TYPE and select NONE CURVE FIT TYPE NONE NONE LINEAR QUAD MORE ELx800 Operator s Manual 48 Chapter 3 Operation 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 HBS AG 1 1 Use the numeric keys to enter the number of any predefined Assay Definition Files stored in the reader s memory or the Options key to advance one assay at a time The cursor is positioned at the first
140. rm the mechanical alignment optical accuracy linearity and repeatability of the instrument e Liquid Testing page 116 uses liquid solutions in a microplate to confirm mechanical alignment optical accuracy linearity and repeatability of the instrument Set the ELx800 to Normal Read Mode when running all verification procedures Chapter 3 Bio Tek Instruments Inc Qualification Procedures 107 System Test IMPORTANT Do not turn on the instrument until the carrier shipping block has been removed Refer to Chapter 2 The System Test is performed automatically whenever the instrument is turned on It can also be performed manually through the ELx800 Main Menu 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 Bio Tek 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 ELx800 in Chapter 2 for instructions 2 From the Main Menu select UTIL The SELECT UTILITY OPTION screen will appear ELECT UTILITY OPTION ESTS SETUP OUTPUT READ 3 From the SELECT UTILITY OPTION screen select TESTS The SELECT TEST screen will appear ELECT TEST YS TEM CHEK SUM CAL PLAT IE 4 Select SYSTEM to run the System Test
141. rmal limit or a cutoff level of 10 U ml The ranges are provided as a guide only ELISA assays are very sensitive and capable of detecting small differences in sample populations It is recommended that each laboratory determine its own normal range based on the population techniques and equipment employed ANA Result Interpretation lt 10 0 Negative gt 10 0 Positive Bio Tek Instruments Inc 177 Programming the ANA Screen Enzyme Immunoassay Kit Transformation and Cutoff From the Main Menu press DEFINE Select the assay name and edit the name if desired At the DEFINE menu STEP COMMENTS To program the reading method press METHOD WAVELENGTH Dual or Single Single MEASURE Wavelength 450 PLATE TYPE 96 To program the plate map press MAP AUTO Auto mapping is normally preferred because it fills in the well IDs logically and automatically after determining which direction to map and how many wells to fill DOWN Maps the wells down the column DOWN Locates replicates in a vertical orientation down the column A01 Begins mapping at well location A01 BLANK MAP AIR Choose to blank on AIR if no blank wells are required NUMBER STDS 00 NUMBER CTLS 03 CONTROL 1 PC CONTROL 2 CTL1 Suggested choice for the cutoff control CONTROL 3 NC NUMBER OF REPLICATES PC 01 CTL1 03 ELx800 Operator s Manual 178 Appendix E Instructions for Programming a New Ass
142. rmat MATRIX to print in a format that resembles the plate type ex 8 x 12 matrix or BOTH e See Appendix C for examples of Reports 7 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 10 Press ENTER to continue The PRINT CURVE FIT screen will appear RINT CURVE FI T9 Y ES N O 11 Select YES to print the standard curve only applies to quantitative assays 12 Press ENTER to return to the SELECT UTILITY OPTION screen Bio Tek Instruments Inc Using the Utility Options 101 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 1 To specify various read time options start at the Main Menu and select UTIL gt READ 2 The PROMPT FOR PLATE ID screen will appear Press ENTER to cycle through the prompt screens If selected at read time gt gt gt gt 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 Rapid Mode or Normal Mode RAPID MODE reads a 96 well plate in approximately 30 seconds single w
143. rogramming the ANA Screen Enzyme Immunoassay Kit 177 Sample Human Anticardiolipin IgG Enzyme Immunoassay Kit 179 Programming the Human Anticardiolipin IgG Enzyme Immunoassay Kit 555220 rci Frau vet erate 184 174 Appendix E Instructions for Programming a New Assay Sample ANA Screen Enzyme Immunoassay Kit _ Transformation and Cutoff Intended Use This assay is designed for the in vitro screening of human serum for the presence of specific IgG antinuclear antibodies ANAs to aid in diagnosing certain systemic rheumatic diseases Sufficient materials are supplied to allow a maximum of 93 samples to be screened in single with a positive cutoff and negative controls Background Antinuclear antibodies occur in a large number of patients with systemic rheumatic diseases These diseases are characterized by the presence of one or more ANAs Sera positive on this ANA kit should be tested for the specific autoantibodies indicative of the various systemic rheumatic diseases Principle of the Assay Microwells are pre coated with purified antigens The prediluted controls and diluted patient samples are added to the wells and autoantibodies recognizing one or a combination of antigens bind during the first incubation After washing the wells to remove all unbound proteins IgG conjugate is added The conjugate binds to the captured human autoantibody and the excess unbound conjugate is removed by a further wash s
144. rotocol 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 of Endpoint or Kinetic Define the wavelength s at which the plate will be read Select a Plate Type from the drop down list Define other reading parameters as necessary Click the Help button for assistance When complete click OK Select Data Read Plate The Plate Reading dialog will appear Enter any comments 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 Bio Tek Instruments Inc Appendix B Using 384 Well Geometry The following describes how to manipulate data received from a plate with 384 well geometry To capture and manipulate the raw data using 384 well geometry you must use KCjunior or KC4 The Optical Density values will be downloaded via the RS 232 port and added to the data logging program KCjUNIOr oa ie ree vaipsarem eade ale dear wu Wan an x ae oa sean Dia Re dasa RE ax 156 Kc4 159 156 Appendix B Using 384 Well Geometry KCjunior KCjunior Bio Tek s data reduction package for Wi
145. shock and fire hazards ELx800 Operator s Manual 20 Chapter 2 Installation Power Up and System Test After you have installed the ELx800 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 6 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 start at the Main Menu and press UTIL gt TESTS gt SYSTEM See System Test and Checksum Test in Chapter 4 or Connecting a Printer to the ELx800 on page 22 for more information ELx800 Main Menu Following successful power up of the ELx800 the Main Menu appears 01 22 PM 01 23 04 DEFINE REPORT UTI L e The Main Menu permits access to all onboard functions See Main Menu in Chapter 3 for more information including a diagram showing the flow of functionality Figure 14
146. 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 Bio Tek Instruments Inc System Startup 43 System Startup To turn on the ELx800 press the on off switch on the right side of the reader s base The ELx800 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 4 for more information e Refer to Chapter 6 Troubleshooting and Error Codes to interpret error codes and Chapter 1 Introduction for information on contacting Bio Tek Instruments Technical Assistance Center TAC ELx800 Operator s Manual 44 Chapter 3 Operation Main Menu Following successful power up of the ELx800 the Main Menu appears 45 AM 01 31 03 DEFINE REPORT UTI L The Main Menu permits access to all reader functions Figure 15 e READ option or press the key labeled READ on the keyboard for plate reading prompts You will be asked to select from a list of preprogrammed assays e DEFINE option Allows the creation of a reading and data reduction protocol You will be prompted to select an assay from an assay defini
147. t levels low electronic noise adequate photodiode sensitivity and overall system cleanliness The testing is designed to ensure that the ELx800 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 internally coded Failure Mode Effects Analysis FMEA criteria established by Bio Tek 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 Bio Tek Instruments Inc Error Codes 141 Error Codes An error code is displayed on the instrument as a four digit id
148. ta 10077 24136 Channel Ref al Noise Max 9878 9980 Noise Min 9877 9978 Delta T 2 SYSTEM SELF TEST Resets Resets Resets Resets Resets Resets 1 Figure 16 Sample output for the System Test sheet 1 of 2 Bio Tek Instruments Inc Qualification Procedures 109 AUTOCAL ANALYSIS Upper Left Corner x 9090 y 10760 ower Left Corner x 9082 y 16280 Lower Right Corner x 412 16282 Upper Right Corner x 416 10762 Delta 1 9090 9082 8 Delta 2 416 412 4 Delta 3 10762 10760 2 Delta 4 16282 16280 2 SYSTEM TEST PASS Figure 17 Sample output for the System Test Autocal Analysis sheet 2 of 2 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 and displays the 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 Press UTIL at the Main Menu At the SELECT UTILITY OPTION screen press TESTS and then CHKSUM at the SELECT TEST screen to run the Checksum Test The information displayed will resemble the following 330202 VERSION OD E CHECKSUM gt The initial checksum test display will show the onboard base code software part number version number and
149. te 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 position 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 specification is applied using Normal Mode and a 96 well microplate t 1 076 0 010 OD from 0 000 to 2 000 OD 405 nm Bio Tek Instruments Inc Liquid Testing 125 Liquid Test 3 for UV Models Only 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 ELx800 at 340 nm and is optional because the reader has good front end linearity throughout its wavelength range Requi
150. tep Substrate is added that causes a blue reaction thereby exposing the bound conjugate and producing an intensity proportional to the concentration of autoantibody in the sample Phosphoric acid is added to each well to stop the reaction This produces a yellow end point color which is read at 450 nm Bio Tek Instruments Inc Materials 175 Materials Supplied Instruction Leaflet Giving full assay details QC Certificate Indicating the expected performance of the batch ANA Coated Wells 12 x 8 well strips coated with purified antigens Type III Wash Buffer 20x Concentrate 1 bottle containing 50 ml of a concentrated buffer for washing the wells Type III Sample Diluent 2 bottles containing 50 ml of buffer for sample dilution Ready to use ANA Positive Control 1 bottle containing 1 8 ml of diluted stabilized human serum Ready to use ANA Cutoff Control 1 bottle containing 1 8 ml of diluted stabilized human serum Ready to use ANA Negative Control 1 bottle containing 1 8 ml of diluted stabilized human serum Ready to use ANA Conjugate 1 bottle containing 12 ml of peroxidase labeled antibody to human IgG Colored red ready to use TMB Substrate 1 bottle containing 14 ml TMB substrate Ready to use Stop Solution 1 bottle containing 14 ml of 3M Phosphoric acid Ready to use Additional Materials and Equipment Not Supplied Automatic Microplate Plate Washer This is recommended however plate
151. ter LCD screen and membrane switch Data can be stored onboard printed and or uploaded to controlling software on a host PC The ELx800 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 The ELx800 is backed by a superior support staff If the ELx800 ever fails to work perfectly please call the Technical Assistance Center or visit Bio Tek s Web site Refer to the end of this chapter for contact information Bio Tek Instruments Inc Hardware Features 3 Hardware Features The ELx800 s hardware features include e Single optics channel e Wavelength range of 400 750 nm ELx800UV model provides extended wavelength range of 340 to 750 nm e A 5 position filter wheel e A 2 line x 24 character LCD display e A membrane keypad with alphanumeric keys e X Y carrier movement e Capability of reading 6 12 24 48 and 96 well microplates ELx800NB model extends read capability to include 384 well and 60 72 96 well Terasaki microplates e External 24 volt power supply which runs on 100 to 240 V 10 0 50 to 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 Trans
152. tion for well A1 is less than 0 017 the well meets the test criteria Linearity Test 1 Foreach ofthe 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 2 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 39 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 e 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 Bio Tek Instruments Inc Chapter 5 Maintenance and Decontamination This chapter contains procedures for maintaining and decontaminating the ELx800 Maintenaricess sso stia sce Ea ir dfe er a EEN pele nics ues ee Dat Roe eee Routine Cleaning ProCe Cure ccccccssccceceecsseessceesseeeeseeeeseetseeeeanes Replacing the BUlb sucseuvexeuvksbuy xia deve vi
153. tion list e REPORT option Allows printing of results and protocol descriptions You will be prompted for the name of a previously run assay with valid data e UTIL option You will be prompted to select options from the list of onboard utilities READY 09 14AM 09 08 04 READ DEFINE REPORT UTIL SELECT UTILITY OPTION TESTS SETUP OUTPUT READ PRINT REPORT RESULT MAP ASSAY LIST SELECT ASSAY NUMBER 53 NAME 53 Bordetella M SELECT ASSAY NUMBER 53 NAME 53 Bordetella M Figure 15 Options available from the Main Menu Bio Tek Instruments Inc Main Menu 45 Important On some Universal Microplate Readers Assay 01 has been designed to allow for quick and simple programming It appears as _Quick Read on the display Most of the options available in Assays 2 55 and described in this section are unavailable for programming within Quick Read You can quickly access the Quick Read assay by selecting READ from the main menu The Quick Read assay default DEFINE settings are shown below and cannot be edited except where noted Method e Single Wavelength 405 nm editable Map e 96 well plate geometry e Blank on Air e Automap e Map starting location A1 e Samples only no blanks standards or controls e Sample count prompted at runtime can be turned off in UTIL READ options ELx800 Operator s Manual 46 Chapter 3 Operation Define The Main Menu option DEFINE allows you to define the
154. trates 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 2 Table 2 Typical Enzyme Substrate Combinations and Stopping Solutions Substrate Stopping Solution Alkaline o nitrophenyl phosphate 3N sodium hydroxide Phosphate Peroxidase 2 2 Azino di ethylbenzothiazoline sulfonic citrate phosphate acid ABTS buffer pH 2 8 o phenylenediamine 0 03N sulfuric acid beta o nitrophenyl beta D galactopyranoside 1M sodium carbonate Galactosidase Bio Tek Instruments Inc Liquid Testing 117 Stock Solution Formulation The stock solution for Liquid Tests No 1 and No 2 may be formulated from the ingredients listed below Solution A or by diluting a dye solution available from Bio Tek 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 Bio Tek wetting agent PN 7773002 e Precision balance with readability of 0 001 g e Weigh boat e 1 liter volumetric flask Preparation of Stock Solution 1 Weigh out 0 092 gram of FD amp C No 5 yellow dye powder into a weigh boat 2 Rinse the contents into a 1 liter volumetric flask Add 0 5 ml of Tween 20 or 5 ml of Bio Tek s wetting agent 4 Make up to 1 liter with DI water cap and shake well e
155. ts affiliated dealers Bio Tek Instruments Inc Contents iii Contents NOEICGS ciues URN NUN a a DAMEN EUR DEN RN CERA DA DN REND AD ii All Rights Reserved uisi eive pa a aka de E XR da e Pag Ra tenant eas Eb a E ad paa ad Rad ii d icis RR CETT TTE LU T Tt NNE OE E ii Restrictions and Liabilities Mm ii Contact InfOrrmalblOri oasis ce dv dh id n nl Od RR CER NU GERD RE EA dn PRO HR D ER EU NE xi Customer Service and Sales xc sn n rd E ADR BL GEGOCE OUR ER EO OR CET TAN xi crJauieJAP ie LEE UMETM xi European Coordination Center essicccadannaniontvassnanserdudadiasiaadel AER FPAADRYTAZA YQ ENRE xi DOCUIMNENE CONVENUIONS sexs ci vce decent eee aic abe eau Eure avian dadeaves csv uda Exe RL Er KE MEE EE xii R vision SEDE V sessen aaa x C3 aso cota dt FR ER DE E Duk PEE Gs de di baa da E aul y cep Ra xiii Intended USe Statement assentiri ER XE EX KEY ERE Xu E RR ER NAE V E RU IO REA DTE RE REA UA XV Quality CONTrOl Mc XV Repackaging and SHIPPING aiias uos iiia 3a YE Rak Ea ue Eras Au Va Va OE AR SE 2R E VAR RES Ea RN VA Ra XV b quiecqe DE xvi Cile e xvi POAC Sy cid Ux a D E RDUM d s OE ODD CE DN CO xo ACUERDA xvi Hazards and PrecautiOlis osi ovxtont a EY UI MO EA O EBORE RR EK RE UA AO HU NO xvii uro A MCEI p eva dius ae dane dette xvii Precautii me xviii Directive 89 336 EEC Electromagnetic Compatibility c sesseeeesse
156. uee eee eee es rus 1089 SAMPLE Los oss ors ue trt vem ss tsm te om ree m a Se re zzr9 202 tee 19s ses ses 159 ware PEER 200 essc osi poder ee Set 2453 Serial 161259 Figure 18 Sample Absorbance Test Plate data sheet Note The Test Plate Analysis report may vary slightly if KC4 or KCjunior are used Absorbance Test Plate Analysis Wavelength 405 Alignment Results B2 0 000 PASS B12 PASS G1 0 000 PASS G11 0 000 PASS Accuracy Results Col F05 G03 STANDARD 0 147 2 279 1 133 DATA 0 139 2 265 1 128 RESULT PASS PASS PASS Repeatability Results Col D04 READ1 0 139 22914 READ2 O l4l 2 9T3 RESULT PASS PASS Figure 19 Sample Test Plate Analysis Report Bio Tek Instruments Inc Qualification Procedures 113 Entering the Absorbance Test Plate Specifications Using the Data Sheet provided with the Absorbance Test Plate enter the calibration values To enter the values into the reader from the instrument keypad 1 Start at the Main Menu and select UTIL 3 SELECT UTILITY OPTION SETUP The EDIT SETUP INFORMATION menu will appear SETUP INFORMATION T IME FILTER MORE 2 Press MORE to display the second EDIT SETUP menu Select CALPLATE to access the CALIBRATION FILTER menu 3 Using the insert packaged with the Test Plate select the filter wavelength 4 Press ENTER The WELL LOCATION CALIBRATION VALUE screen appears AVELENGT Hi 0 5
157. uments Inc Define Method Map Formula and Curve 77 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 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 At the SCOPE VARIABLE selection screen choose OD and 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 TVAR is available as a MAP option ELx800 Operator s Manual 78 Chapter 3 Operation Cutoff Formulas A cutoff formula calculates a cutoff value that is used for classifying samples See Formula Type on page 70 for instructions on selecting an assay and accessing the Cutoff formula definition scr
158. ures Qualification Procedures You may use the tests outlined in this section to confirm initial and ongoing performance of the ELx800 Set up the ELx800 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 e Note An Installation Operational Performance Qualification IQ OQ PQ package PN 7330538 for the ELx800 is available for purchase Contact your local dealer for more information Your ELx800 reader was fully tested at Bio Tek 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 confirms that the light levels at all installed filter wavelengths and electronic noise meet factory acceptance criteria The Checksum Test page 109 compares the onboard software with internally recorded checksum values to ensure that no corruption has occurred e The Absorbance Plate Test page 110 uses Bio Tek s Absorbance Test Plate to confi
159. 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 limits Use Tween 20 if possible or some other wetting agent to normalize the meniscus Some solutions develop menisci over a period of several minutes This effect varies with the brand of microplate and the solution composition As the center of the meniscus drops and shortens the light path the density readings change The meniscus shape will stabilize over time Bio Tek Instruments Inc Recommended Test Schedule 105 Recommended Test Schedule The schedule shown in Table 1 defines the factory recommended intervals for performance testing for a microplate reader used two to five days a week Note The risk factors associated with your tests may require that the Performance and Operational Qualification procedures be performed more or less frequently than shown below Table 1 Recommended Test Schedule Installation Performance Performance Operational Qualification Qualification Qualification Qualification Monthly Semiannually Initially and Annually Run Liquid Test 2 ONLY if you do not have an Absorbance Test Plate If you run Liquid Test 2 you do NOT have to also run Liquid Test 1 ELx800 Operator s Manual 106 Chapter 4 Performance Verification and IQ OQ PQ Proced
160. value entry for the next standard e f 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 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 Range 6 Well A01 B03 12 Well A01 C04 24 Well A01 D06 48 Well A01 F08 96 Well A01 H12 96H A01 H12 96M A01 H12 ELx800 Operator s Manual 64 Chapter 3 Operation Reuse of Standard Curves The ELx800 has the ability to reuse a standard curve that has already been established Limitations of the Reuse of Standard Curves Standard curves can only be reused in assay positions 31 through 55 Each of these positions can only store one standard curve Standard curves cannot be reused on panels see page 85 for Panel Definition Standard curves will be stored in memory with the Assay Name Standard Concentrations Replicate Counts and Optical Density values for each standard replicate Stored standard curves can only be reused for the assay that the
161. washing can be performed manually Plate Reader Capable of measuring optical densities at 450 nm referenced on air Distilled or Deionized Water This should be of the highest quality available Calibrated Micropipettes For dispensing 1000 100 and 10 ul Multichannel Pipette Recommended for dispensing 100 ul volumes of conjugate substrate and stop solution Glass Plastic Tubes For sample dilution ELx800 Operator s Manual 176 Appendix E Instructions for Programming a New Assay Quality Control and Results Quality Control For an assay to be valid all the following criteria must be met e Cutoff as well as Positive and Negative controls must be included in each run e The OD of the cutoff and the ANA result of the Negative and Positive Controls should be in the ranges specified on the QC Certificate e For example The absorbance of the Positive Control must be greater than 1 200 OD The absorbance of the Negative Control must be less than 0 300 OD Note If the above criteria are not met the assay is invalid and the test should be repeated Calculation of the Sample Results Use the following formula to calculate the ANA result for each sample Control or sample OD x 10 Control or sample value U ml Cutoff control OD Expected Values The normal range was determined on serum from 200 normal adult blood donors The cutoff control has been set at a point equivalent to the upper no
162. x E VENTER AER V eria A EY nce Rat Decontamination Procedure csssssssesee nennen 130 Chapter 5 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 disconnect the reader from the power supply for all cleaning operations Warning Internal Voltage Always turn off and wet cloth Do not allow the cleaning solution to run into the interior of the instrument If this happens contact the Bio Tek Service Department Do not immerse the instrument spray it with liquid or use a 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 Purpose Exposed surfaces may be cleaned not decontaminated with a cloth moistened not soaked with water or water and a mild detergent Bio Tek Instruments Inc Routine Cleaning Procedure
163. x800 supports seven different curve fitting methods linear quadratic cubic 4 P 2 P cubic spline and point to point ELx800 Operator s Manual 82 Chapter 3 Operation Linear curve fit A simple best fit straight line is plotted using the values of the standards 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 Cubic curve fit A curve fit that uses the equation ax bx2 cx 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 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 logistic equation is algebraically transformed to a simpler form in which experimentally determined values are used for the responses at concentrations of zero and infinity Note Uses linear axis type only 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 4 Parameter Logistic or 4 P A curve fitted to the standard values which is characterized
164. xix Directive 73 23 EEC Low Voltage eccle ntes rnt anra pre rn nah re aacH ER aaS EE xix Directive 2002 96 EC Waste Electrical and Electronic Equipment xix Directive 98 79 EC In Vitro Diagnostics sexe oh yx Su ExRen eu xe Ex CER EAPERRERFEAEAYR xix Electromagnetic Interference and Susceptibility sees XX User Safety de e cc RE EE XX MISIT AESLARD Pe PME HER nC xxi Warrahty e c xxiii Registration C abt uenis aui cca RE RA du de duda Ve EM Edad dd Lap CX LR ORA RARE IR Ra E ON xxiv ELx800 Operator s Manual iv Preface Chapter 1 Introduction acisscoruer uuu ERA DESMREREAKEFEMSENEUREASRESNRER ER E RC RENNEEE 1 Introducing the ELx800 Automated Microplate Reader eseeeeeesee 2 Hardware Features cise ere e nr err eR E E EREEX NA REXEEXXXEREEFARREREENEERKESEEIRNE 3 Software Fedt resss cse IRI rex E PE Iva E UP E QUE 3 Package Contents inso c Ea a CURAR IMMER ahd nanan EE SR E ESATA REIR a aA 4 Optional ACCOSSOFIBSa D stereregvx ori eivasve VITIS Pha E DERART TUER E Rd NER 5 SPECHICATIONS essasuvanacs eva Fia ora rk olo e ael r eR TF p Za a 3 Y Eit aD EL Rn kc a 6 Standard Models seit vds eu tee ex xuR EE E CD REND EX REDE UN UAT UIN CU ES TN DE CEN 6 Ultraviolet UV Model v dcus noe ecu d E e e Ix Or vox va EC REDE DUE Tu 7 Narrow Beam NB Model ssssssseene memes eee ena aaa na 8 Technical SUpport zuussssaspeiezsesa soya Veka SU R ox EREEVERVE AVES DRA E SEX TOR ERE
165. zards associated with the instrument However the operator should be aware of certain situations that could result in serious injury these may vary depending on the instrument model Bio Tek Instruments Inc Hazards and Precautions xvii Hazards and Precautions Hazards cord 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 A Warning Power Rating The instrument s power supply or power 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 Liquids Avoid spilling liquids on the instrument fluid seepage into internal components creates a potential for shock hazard Wipe up all spills immediately Do

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