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ELx808 TM Absorbance Microplate Reader

1. Table 4 6 Instructions for Periodic Lubrication of ELx808 Robotic Units Component Location Procedure A Lubricate the underside of the bracket B Lightly lubricate the shaft indicated C Apply a heavy coating of lubricant to both sides of the motor shaft Run the motor shaft back and forth through the motor to ensure that the internal drive nut is heavily lubricated Use the robotic motor adjustment procedure described on the next page to work the lubricant into the internal drive nut D Lightly lubricate the surface of the roller E Lightly lubricate the underside of the hook in bracket 3 Reattach the top shroud Figure 4 4 Location of bracket motor shaft and main PCB motor assembly 4 30 Performance Verification and IQ OQ PQ Tests Figure 4 5 Roller surface and hook in bracket Robotic Motor Adjustment Procedure After lubrication reassembly the robotic door needs to be set to the open and closed positions Press the following keys to adjust the door At the Main Menu Press the UTIL soft key Press the SETUP soft key Press the hidden key between the Main Menu and Previous Screen keys You should see the Door Adjustment screen Press the CLOSE soft key Press the UP soft key until you just see the door move up You may have to press the key many times before the door moves When the door does move press the DOWN soft key 4 times This positions the door lift ram just behind
2. 0 4 0 2 if original solution is 2 0 at 200 yl Note The choice of dilutions and the absorbance of the original solution can be varied Use Table 4 4 as a model for calculating the expected absorbances of a series of dilutions given a different absorbance of the original solution PLATE PREPARATION 1 Pipette 200 ul of the concentrated solution from Tube 1 into each well of the first column A1 to H1 of the microplate 2 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 4 22 Performance Verification and IQ OQ PQ Tests LINEARITY TEST A 1 Read the microplate prepared on the previous page five times using Normal mode dual wavelength at 450 nm with 630 nm as the blank Note Do not discard the plate you will use it again for Test C Print the ten sets of raw data or export it to an Excel spreadsheet using KC4 or KCjunior The mathematical computations described below may then be performed and the template kept for future data reduction Calculations 3 Calculate the mean absorbance for each well and then average the means for each concentration 4 Perform a regression analysis on the data to determine if there is adequate linearity For example using Microsoft Excel e na spreadsheet create two columns labeled X and Y Enter the actual absorbance values in column X Enter the expecte
3. nein ic 3 9 Single or Dual Wavelength 5 see ei rece ee ce e beds 3 9 MEAS Selection viii re Tee ean ween ae ee 3 10 Preface Number of Kinetic Reads Kinetic Duration SelectiON oocccccccccononocnninnincnnnnn o 3 10 Kinetic InterVal C 3 11 Kinetic Number of Reads iieri enirn three nennt rine nnt 3 11 Kinetic Duration edite a ee e dee uz 3 12 Shake Mode Selection 5 eere ec ote e Ed ipee eo e eo Eee e e eas 3 12 Shake TIMES Lain E ued ii Rt nte Ma eed 3 13 Shake Speed 22 tone eie imi IE 3 13 Kinetic Data Analysis Selection eseee emm 3 14 Number of Kinetic Points Selection sseeeee 3 14 Onset OD Select curia ica 3 15 Linear Scanning ere oia 3 15 MAP it AA elles t Ta aa ia ele Ge 3 16 Map Generation a eene eene rec orte deett 3 17 Mapping A emen emm nennen nnne 3 18 Replication Direction errem rettet He rrr PL ener 3 18 Examples of Mapping Directions sessee mm 3 19 Start Mapping at Well Location sssen mme 3 20 Selecting a Blank Map occiso ad oe rociar 3 20 Blank Map Definitions oret rada 3 21 Constant Blank Value Entry sse emen nennen 3 22 Number of Blanks 91 2 rine che eet e Ue odere pipe ens 3 22 Selecting a Blank Location 11 reete ennt trente rie 3 23 Number of Standards
4. Important The pin indicating the intended voltage should protrude through the cover Do not power up the instrument until the voltage input range to be used is indicated correctly by the pin ELx808 Operator s Manual 2 7 Reconfiguring or Replacing Fuses Either USA or international style fuses are installed in the reader s power entry module The reader s fuses are configured at the factory prior to shipping Use the procedure described below if you need to reconfigure or replace fuses Note A failed fuse is usually an indication of another problem that a new fuse is not likely to repair Contact Bio Tek s Technical Assistance Center if the fuse replacement fails to rectify the problem See Chapter 1 page 1 7 for contact information The fuse module has two fuse configurations e 100 120 V USA A fused hot AG 1 5 A 250 V Slo Blow PN 46024 e 230 240 V international Both hot and neutral fused with a 5 x 20 mm 0 63 A 250 V Slo Blow PN 46038 To reconfigure fuses 1 Unplug the reader and remove the power cord 2 Insert the needle nose pliers into the two holes on the cover of the power entry module Lift the cover open and swing it back into the socket Note If the cover pops out of the instrument instead of swinging back and the fuse holder is attached to the cover your instrument has an older style power input module Turn to Appendix F now 3 Using the pliers gently pull the fuse holder out of the power
5. Appendix B B 7 Computer Control Command Set B 8 A command from the computer to the reader consists ofa single ASCII character and in some cases subsequent argument data Upon receipt of a valid command character the reader returns an lt ACK gt character Some commands also return response data to the host computer Upon completion of command processing the ELx808 transmits a status string to the computer When the reader sends data to the computer the data is sent first then the status string While awaiting a command the ELx808 responds to nulls or other unexpected characters by clearing its input buffer and transmitting a lt NAK gt Therefore if valid commands are preceded by invalid characters they may be missed Refer to Table B 2 for the ASCII Control Characters used in the computer control protocol All ASCII character strings representing numbers or names are transmitted most significant digit or letter first Data values not indicated as ASCII are treated as binary integers and are transmitted least significant byte first Some commands described here may not be available with older reader basecode versions prior to v3 00 indicated if the Check Feature command returns a lt NAK gt These commands are marked with a Table B 2 ASCII Control Characters Used in Computer Control Protocol ASCII Hex Decimal Control Reader Code Code Code Code lt gt NAK Negative 15 21 U gt acknowledge c o
6. D Important Be sure to replace the seven perimeter screws they increase the interference In addition the screws MUST be installed to hold the shroud in place if you plan to ship the instrument e To remove the filter wheel grasp the center hub of the filter wheel and pull it toward the lamp The wheel should easily disconnect from the hub Lift the wheel from the instrument e 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 ELx808 Operator s Manual Check Adjust the Line Input Voltage Range 2 6 The ELx808 is equipped with a four voltage range power entry module This power entry module located on the right side of the instrument can be adjusted for 100 VAC 120 VAC 230 VAC and 240 VAC voltage inputs The setting can be determined visually by observing which indicator hole on the power entry module has a pin protruding from it Check the pin now to make sure the setting is appropriate for your location The following instructions describe how to adjust the input voltage range if necessary The figure below illustrates the power input module Note Small needle nose pliers are required for this procedure ON OFF switch Voltage selector card Fuse holder Power cord Figure 2 3 Power entry module accessing the voltage selector card and fuse holder 1 Unplug the reader and remove th
7. e Select YES to exclude the replicate from curve fit calculations e Select NO to retain the replicate e Press ENTER to advance to the next replicate ELx808 Operator s Manual 3 55 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 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 D Note Each curve fit type requires a minimum number of standards for Printing Results After the assay 1s selected and standard outliers are edited 1f necessary the results report can be printed PRINT RESULTS 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 3 56 Operation Map e Select REPORT at the Main Menu and then select MAP SELECIT ASSAY NUMBER NAME HBS AG e Use the keyboard to type the assay number or the OPTION key to cycle through the list of available assays Press ENTER to enter the assay and begin printing the map of programmed well locations in the selected geometry Assay e Select REPORT at the Main Menu and then se
8. final status string 5 ELx status mode only Temperature response included only if requested Format is three ASCII bytes scaled up by ten 1 e 275 correponds to 27 5 degrees C See the Include Temperature Response command Data is returned in the order wells are specified If fewer than eight wells are to be read each well s unused row and column must be replaced with 00 Data is always returned for eight wells regardless of the actual number specified See the Read Plate command for details concerning results data format and error handling and status string information This command always returns data using the ELx format 4 GET BASECODE VERSION e B 10 This command returns a string containing the part number and version number for the instrument downloadable basecode software host valid limits response e no arguments lt ACK NAK gt reader response protocol part number 7 intermediate string 10 version code 7 status string 5 0000000 9999999 Version x xxyyy The version number is indicated by the first four characters of the version code x xx The remaining characters in the version code yyy may either be blank or represent a specific beta or prototype release code Computer Control INCLUDE TEMPERATURE RESPONSE i This command causes the current temperature to be included in the results response for each plate read using the
9. Added IQ OQ PQ procedures and updated liquid testing information Chapter 4 Revised lamp replacement procedure page 2 15 Preface Updated contact information in Notices page 111 Added Document Conventions page vi Updated Warnings section pages vii and viii Updated Electromagnetic Compatibility section pages ix and x Added the following safety symbols and text gt Consult instructions for use page xii gt In vitro diagnostic medical device page xii Separate collection for disposal of electrical and electronic equipment page xiii Preface Revision History Cont Rev Date J 11 03 K 4 05 ELx808 Operator s Manual Changes Preface Cont Expanded the Intended Use Statement page xiv Chapter 1 Updated contact information in Technical Support Removed About This Manual section page 1 5 Added Absorbance Test Plate to the Optional Accessories list page 1 8 Clarified lamp replacement procedure Chapter 2 pages 2 14 to 2 16 Clarified description of cutoff formulas Chapter 3 pages 3 42 to 3 44 Chapter 4 Changed title to Performance Verification and IQ PQ OQ Tests Added IQ PQ OQ test procedure information Clarified procedures for liquid tests Revised decontamination instructions and added cleaning procedure Appendix A Added KC4 startup information to Appendix B Added new Appendix E with two examples of assay kit instru
10. DEFINE Bold Text in COURIER font represents menu options as they appear on the display of the ELx808 Note Bold text is primarily used for emphasis D This icon calls attention to important information Preface Warnings e Operate the instrument on a flat surface away from excessive humidity 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 hazards 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 Hazards A Warning Power Rating The instrument s power supply or power 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 Warning Electrical Grounding Never use a two prong plug adapter to connect primary power to an instrument or to a power supply Use of a two prong adapter disconnects the utility ground creating a severe shock hazard Always connect the system power cord or power supply directly
11. Preface Qualification ProCedures occccccccccococococonononononononononononononononononenanononononononononononononononononaninonas 4 5 Initiating Tests Via the Utility Option seenemm mme 4 6 SYSTEM SystermTest cia dd dee 4 7 CHKSUM Checksum Test ena ceti eL e ede e e ee cd 4 9 CALPLTE Absorbance Plate Test eene 4 9 Absorbance Plate Test cis 2 unm D Dr REFER cede 4 10 Requirements un unen NB IER i 4 11 Entering the Test Plate Specifications eseee een 4 11 Running the Absorbance Plate Test eee 4 12 Repeated Test Fall res x ic iii siria 4 15 Empty Carrier Test 5 nin Ho dee te ue ERU Ep i edere cen beets 4 16 Liquid Testing cocos de eiie geteilt deed decide Eae de dede decre 4 17 Stock Solution Formulation esese eene een nennen 4 18 quid desd tte e E e E A aras 4 20 Liquid Tesla Ne o E TEA se 4 22 quid Test m latin caves e oie Dotes 4 25 Periodic Maintenance for the ELX808 ooonooccccnnocccocococcconononcnononcnnonnnnnnnn eene 4 28 Cleaning the Lamp Contacts ssssssssseeeeene nennen ener 4 28 Inspecting and Cleaning the Wavelength Filters ee 4 29 Periodic Maintenance for ELx808 Robotic Units senem 4 30 Replacing the Lamp eerte ta tae 4 32 Appendix A Decontamination Leeuee eese ueeeueeeueeeeeeseee A 1 Appendix B C
12. e Turn on the printer Figure 2 5 Serial and Parallel ports ELx808 Operator s Manual 2 11 Printers The ELx808 s parallel port LPT1 allows connection to compatible printers The reader 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 call B1o Tek s Technical Assistance Center or visit our website refer to Chapter 1 for contact information Cable A printer cable is supplied with the reader If this cable becomes lost or damaged Bio Tek offers a variety of printer cables Contact your authorized Bio Tek dealer for information on cable prices and availability Parallel Port Pin Definition The table that follows illustrates the pin definitions for the reader s 25 pin socket female D sub Parallel connector Table 2 1 Parallel Connector Pinouts Parallel Port Pinout Pin signal Pin z fo h b m CN m EC s m s s m EC s m n CON ta E m msy EC Signal so D D Uo GN GN 6 6 5 6 o Nc fs oo bape Installation Turn on the Reader and Run a System Test If all of the required steps preceding this one have been performed successfully locate the reader s power switch on the right side and turn on the reader The ELx808TM will automatically perform a System Test The System Test conducts a series of tests at each wavelength defined in the filter table to confirm adequ
13. ATOD INIT ERR 1C00 A D calib STBY transition not detected RESULTS DATA ERR 1D00 results data error CLOCK ERR 1E00 error in clock communications OVERLAP ERR 1F00 bandpass overlap in filterset BARCODE ERR 2000 no valid barcode detected INVALID PARAM ERR 2100 invalid parameter value selected PMT ERR 2200 PMT test signal too high test type LAMP ERR 2300 lamp control failure test type SENSOR POS ERR 2400 test sensor position incorrect lt motor gt FLASH MISS ERR 2500 motor went by flash location too soon XY LIMIT ERR 2600 physical limit exceeded for area scan request PANEL METHOD ERR 2700 lt assay gt method doesn t match first panel assay MOTOR TIMER ERR 2800 lt motor gt timer not available Test Type Codes lowest digit in returned error code FAIL POWER 5V el 5V power failed FAIL POWER 24V 2 24V power failed Motor Codes lowest digit in returned error code Carrier X Axis 0 Filter Wheel Y Robotic Door 2 R instruments only Incubator Codes second lowest digit in returned error code Range Error 0 Thermistor Error Y A D Error PA Affected zones are encoded in the lowest digit returned one bit per zone Data Flash Codes 2 lowest digit in returned error code Readback Error 0 data read back didn t match data written Copy Error Y final data readback didn
14. PRINT CURVE FIT N O e The current selection is displayed on the top line of the screen To change the report option press the soft key under YES or NO The display updates to reflect the selection e Choose YES if there are quantitative assays defined on board and you wish to print the curve e Press ENTER to return to the SELECT UTILITY OPTION screen ELx808 Operator s Manual 2 17 READ Options Read Speed 2 18 At the SELECT UTILITY OPTION screen press the soft key beneath READ to set up Reader Prompt preferences Select YES to prompt the operator to enter identifications and sample counts before a microplate 1s read The prompts are shown below Press ENTER after each selection to advance the display Prompt for Plate ID allows the user to enter an alphanumeric name of up to 10 characters Select YES if to present this prompt at run time Prompt for Sample ID allows the user to identify samples with a 4 character alphanumeric name The starting ID 1s entered and automatically incremented by the software Select YES to present this prompt at run time PROMP T SAMPLE COUNT NO PROMPT SAMPLE COUNT allows the user to enter the total number of samples on the plate at runtime only those sample results will be printed Select YES to present this prompt at run time RAPID MODE reads a 96 well plate at 6 second intervals with wavelengths higher than 400 nm Selecting NO means a plate with single wavel
15. 608 690 693 590 614 620 553 636 623 Delta 26359 51004 48565 53714 48193 45715 51326 49018 44906 Filter 490 Gain 3 12 Channel Ref 1 2 3 4 5 6 7 8 Air 27535 51422 49253 54406 48932 46038 51523 49924 46583 Dark 512 579 581 497 517 522 467 535 525 Delta 27023 50843 48672 53909 48415 45516 51056 49389 46058 Filter 550 Gain 3 20 Channel Ref 1 2 3 4 5 6 E 8 Air 27407 51254 49152 54366 48956 45632 51410 49895 47098 Dark 522 591 593 506 527 532 476 546 535 Delta 26885 50663 48559 53860 48429 45100 50934 49349 46563 Filter 630 Gain 10 24 Channel Ref 1 2 3 4 5 6 7 8 Air 27677 50381 48935 53521 48494 45416 51083 49030 46366 Dark 1497 1718 T25 1448 1515 1529 1349 1574 1539 Delta 26180 48663 47210 52073 46979 43887 49734 47456 44827 Filter 340 Gain 51 20 Channel Ref 1 2 3 4 5 6 7 8 Air 27559 49117 43614 49246 43079 44183 46268 45204 36638 Dark 7268 8371 8403 7023 7353 7428 6534 7652 7473 Delta 20291 40746 35211 42223 35726 36755 39734 37552 29165 Channel Ref 1 2 3 4 5 6 y 8 Noise Max 5206 5993 6018 5029 5265 5320 4680 5480 5354 Noise Min 5204 5992 6014 5028 5263 5318 4678 5479 5350 Delta 2 1 4 1 2 2 2 I 4 01 14PM 01 01 04 INCUBATOR SELF TEST Temperature Setpoint 37 0 Current Average 37 0 A D Test PASS Zone 1 37 1 Min 37 1 Max 37 1 Range PASS Thermistor PASS Zone 2 37 1 Min 36 9 Max 37 1 Range PASS Thermistor PASS Zone 3 37 0 Min 36 9 Max 37 0 Range PASS Thermistor PASS
16. ELx808 Operator s Manual 4 9 Absorbance Plate Test The Absorbance Plate Test also referred to as the Calibration Plate Test can be used to conduct several useful tests in a short period These tests are Mechanical Alignment Accuracy and Linearity Reproducibility and Channel to Channel uniformity To run the test you will need Bio Tek s Absorbance Test Plate PN 7260522 or 9000547 with its accompanying Data Sheet The Test Plate allows for the comparison of the reader s optical density measurements and mechanical alignment to NIST traceable values Specific standard calibration values shown on the Data Sheet must be entered for each wavelength to be tested Note If a Test Plate is not available an alternative method that may be used to determine relative accuracy repeatability and linearity is Liquid Test 2 described on page 4 22 The Absorbance Plate Test 1s comprised of the following subtests the data and results of which can be reviewed on the Absorbance Plate Test report see page 4 13 for a sample report e Mechanical Alignment The Test Plate has several groups of precisely machined holes to confirm the mechanical alignment of different microplate readers The amount of light that shines through these holes is an indication that 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 reader may be out of alignment
17. Zone 4 36 9 Min 36 9 Max 36 9 Range PASS Thermistor PASS AUTOCAL ANALYSIS Upper Left Corner x 4968 y 0 Lower Left Corner x 5754 y 0 Lower Right Corner x 14440 y Upper Right Corner x 13652 y Delta 1 4968 5754 786 Delta 2 13652 14440 788 SYSTEM TEST PASS Figure 4 1 Sample System Test Report Performance Verification and IQ OQ PQ Tests CHKSUM Checksum Test The Checksum Test compares the basecode software to the internally recorded checksum values to ensure that the programming has not been corrupted To verify the checksum revisions and version of software currently loaded onto your reader select CHKSUM from the TESTS submenu If an error is detected the display will show an error code Note This test also runs automatically when the reader is turned on When the CHKSUM Test is selected the software versions and the software s checksum appear on the display as shown in the example below Software P N Software Version 7330203 Version 3 14 Code Checksum 6B87 The second screen shows Configuration P N Configuration Version 7330203 Version 2 82 CALPLATE Absorbance Plate Test The Absorbance Plate Test also referred to as the Calibration Plate Test confirms the alignment accuracy linearity and repeatability of the ELx808TM To initiate this test select CALPLATE from the TESTS submenu More information on the test is provided in the next section Absorbance Plate Test
18. coito aer 3 23 Number of Standard Replicates oonooccccnnonicinnccccnnnonnncnonancnn nono nn iien naar nnnrnnnnnno 3 24 Average Standar uti 3 24 Standard Concentration ieni etario 3 25 Reuse of Standard CUIVES comicios 3 26 Number of Controls 2 2 reete id 3 27 Control Ty Pe C A A aa 3 28 Number of Control Replicates oooooononccnnnociconoccccnnnoncnononancnn non cnn emen 3 28 Location OF COS ect ER RETRO Set Pe ent 3 29 Valid Well Eocations eite tenete tati 3 29 Number Of esr ecc 3 29 Number of Sample Replicates sseeeeeeeeeme enm 3 30 Sample Location ib da 3 30 FORMULA 200 A bina ee eee eins 3 31 Validation Formula Examples seen eme nene 3 31 ELx808 Operator s Manual xvii xviii Formula Pype Sci ii 3 32 Validation Type Selectlon eene nene te tentent entendi nnn rines 3 33 Formula Entire e AUI E R a eei lads tut 3 34 Number of Required Controls Blanks eeee eee 3 36 Cutoff Formulas 2 es we ad e ee nes 3 37 Greyzone EDI nze iine PE ROI UH DH DE aa nes 3 38 Positive Negative Calls for Cutoff sse 3 38 Examples iati ici 3 39 Transformation Formulas sesseeee eene eene nnne 3 40 Transformation Formula Definition eeeeeeee enm 3 41 Transformation Scope Variable sseeeseeeene enne 3 42 CURVE M 3 43 Curve FIL eere ni eee
19. soft keys located below the on screen menu options READ DEFINE REPORT and UTIL are activated and may be selected 01 30PM 10 09 0 DEFINE REPORT UTI Figure 3 2 Main Menu screen 01 30PM DEFINE REPORT Figure 3 3 Main Menu screen in I Incubator instruments Note The temperature indicated on the display of incubated models is the actual averaged temperature of the incubator s four zones The applied setpoint of the last assay is used To adjust the temperature a new setpoint value must be assigned to the assay before running ELx808 Operator s Manual 3 3 Press the soft key that corresponds to a displayed menu option to activate that option e READ option Initiate a plate read Or press the key labeled READ on the keyboard for plate reading prompts You will be prompted to select from a list of available assays e DEFINE option Create a reading and data reduction protocol You will be prompted to select edit an assay and then define its various parameters e REPORT option Print results reports and protocol descriptions For results reports you will be prompted to select a previously run assay with valid data e UTIL option Access various onboard utilities used for configuring and testing the reader and simple assay 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 wi
20. water to adjust the stock solution Note that too small a well volume may result in increased pipetting related errors ELx808 Operator s Manual 4 19 Liquid Test 1 Note A 96 well flat bottom microplate is required for this test Corning Costar 3590 is recommended Use a new microplate any fingerprints or scratches may cause variations in the turnaround reading running in Rapid mode To check this select UTIL gt READ and then cycle D Important Before running the liquid tests ensure that the reader is not through the prompts until READ IN RAPID MODE is displayed Set to NO 1 Using a freshly prepared stock solution A or B see page 4 18 prepare a 1 2 dilution using deionized water one part stock one part deionized water the resulting solution is a 1 2 dilution gt The concentrated stock solution should have an optical density of approximately 2 000 OD or lower 2 Pipette 200 ul of the concentrated solution into the first column of wells of the microplate 3 Pipette 200 ul of the diluted solution into the second column of wells 4 Readthe microplate five times at 405 nm using Normal mode single wavelength no blanking Normal plate position 5 Without delay rotate the microplate 180 so that well A1 is now in the H12 position Read the plate five more times Turnaround plate position 6 Print the ten sets of raw data or export to an Excel spreadsheet using KC4 or KCjunior M The m
21. 1 General Requirements gt EC Directive 73 23 EEC e EN 61010 1 1990 Safety requirement for electrical equipment for measurement control and laboratory use Part 1 General requirements Including Amendment No 1 ELx808 Operator s Manual ix 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 EN 50081 1 1992 and IEC 61326 1 1998 EN 55022 1995 Class A Immunity EN 50082 1 1992 and IEC 61326 1 1998 IEC 801 2 Electrostatic Discharge EN 61000 4 3 Radiated EM Fields EC 801 4 Electrical Fast Transient Burst IEC 1000 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 Safety requirement for electrical equipment for measurement control and laboratory use Part 1 General requirements Including Amendment No 1 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 gt 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 iii for contact information Directive 2002 96 EC Waste Electrical and Electronic E
22. 1007 high speed sweep reads 1008 partial plate reads 1009 incubation 1010 sensitivity response 1011 multi wavelength read format 1012 serial read mode selection 1013 ELx format status response 1014 serial only instrument 1015 basecode checksum 1016 barcode scanning lt ETX gt reader response protocol feature available 1 miscellaneous additional data 16 status string 5 response lt ACK NAK gt associated commands legacy V not available legacy M legacy V legacy not available Te legacy I readers only P i not available Pr If the z is acknowledged ACK the instrument shall then return 1 if the feature is available 0 if not If the z command is not recognized indicated by a NAK response to the z it means the instrument basecode version was released prior to implementation of this command along with the other commands marked as such with a In this case if the feature is marked in the table above as legacy the indicated commands and functions are and always have been available anyway Computer Control Using the Stop Key to Halt Plate Reads Pressing the STOP key on the reader while a computer control initiated plate read is in progress causes the following to occur 1 The read process is halted and all axes ar
23. 5 respectively In normal pregnancy the levels are higher still at 17 096 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 lt 11 GPL U ml Negative result gt 11 GPL U ml Positive result Appendix E E 11 Programming the Anticardiolipin IgG Enzyme Immunoassay Kit Standard Curve and Cutoff D Important The ELx808 user is responsible for programming the reader properly according to their specific kit instructions and for verifying that the calculations are performed correctly If you are not familiar with programming formulas onboard the ELx808 refer to Formula Entry in Chapter 3 From the Main Menu press DEFINE Enter the assay number and edit the name if desired At the define menu follow the steps below l STEP To program the reading method press METHOD READ TYPE Endpoint DELAY FIRST READ IF INCUBATOR MODEL INCUBATION TEMP Ambient or Temperature DELAY FIRST READ WAVELENGTH Single or Dual Single MEASURE Wavelength s to use 450 SHAKE First Every or None To program the plate map press
24. KEYS 1 2 or 3 to select the BLANK MAP type above the soft key The display updates to reflect the selection e Press SOFT KEY 4 to access MORE options ROW or COLUMN and P ACROSS or P DOWN e Press ENTER to save the well location and advance to the next screen BLANK MAP DEFINITIONS e 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 e FULL enables a single blank well or an average of blank wells to be subtracted from the whole plate e CONST Constant allows entry of a user specified absorbance value This value will be subtracted from each well on the plate Use a blank value from the first plate or a blanking plate to save space on subsequent assay plates e ROW enables a single blank well or an average of blank wells to be selected for each row The blank or average will be subtracted from each well in the row Use manual mapping to position blanks controls standards and samples e COLUMN enables a single blank well or an average of blank wells to be selected for each column The blank or average will be subtracted from each well in the column Use manual mapping to position blanks controls standards and samples e P DOWN enables a blank in every even numbered column to be subtracted from the well to the left of it in every odd column Manual mapping is recommended to set up the appropriate map by pl
25. Note If extrapolation is chosen for the Point to Point curve fit unknown 3 46 Operation Panel Assays A Panel assay is a collection ofup to 8 assays to be run on one plate e The most common reason to use a Panel assay is for confirmatory tests based on a screening test in clinical applications e Only one panel assay can be defined on the reader at any time e The assays specified within the Panel must be predefined in any of the assay positions 1 55 e The assays specified within the panel must all use the Endpoint read method e The assays specified within the panel must all read at the same wavelength s e Any curve fit type formulas or standard concentrations previously defined for each assay will be used when the assay is selected for a Panel e Panelassays cannot reuse standard curves e The type and number of controls blanks standards and replicates in the assays chosen for the Panel will be copied into the Panel definition To change any of the map or assay parameters in the Panel they must be changed in the predefined assay first e Consider printing a Map Report for each assay that will be included in the panel for use with mapping the Panel To create a panel assay start at the Main Menu select DEFINE then choose assay number 99 Enter the panel assay name e The default name is PANEL e Use the ALPHA and NUMERIC keys to update the Assay name if desired e Press ENTER to continue T
26. OD from 0 000 to 2 000 OD 9340 nm 3 For each well compare the standard deviation calculation in step 1 with the allowed deviation calculation in step 2 The standard deviation should be less than the allowed deviation For example Five readings in well A1 of 0 802 0 802 0 799 0 798 and 0 801 result in a mean of 0 8004 and a standard deviation of 0 0018 To calculate the allowed deviation for well A1 multiply the mean by 1 0 and add 0 005 0 8004 0 010 0 005 to get 0 013 Since the standard deviation for well A1 is less than 0 013 the well meets the test criteria Repeatability Specification 1 0 0 005 OD from 0 000 to 2 000 OD 9340 nm LINEARITY TEST B 1 For each of the three Test Solutions 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 gt Data Analysis gt 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 Note If the Data Analysis command is not available on the Tools menu you may need to install the Analysis ToolPak in Mic
27. OF CONTROLS Use this option to enter the location of controls in the assay The displayed location field can only be edited if manual map generation was selected CONTROLIH1 LOCATION TY PE PC REP 1 A0 1 e Press the CLEAR key to clear the value on the display e Use the NUMERIC and ALPHA keys to enter values for well locations VALID WELL LOCATIONS For all prompts of a well location only the ALPHA keys are active for the first character and NUMERIC for the second and third characters e Press the MAIN MENU and PREVIOUS SCREEN keys to move back and advance through the menu structure NUMBER OF SAMPLES This option allows you to enter the number of sample groups that will be used in the assay Any previously defined value appears on the display If the number of samples is altered the number of replicates for the sample reverts to a value of 1 ENTER NUMBER O F SAMPLES e Use the NUMERIC keys to enter the number of samples in the assay The range is 0 to the number of undefined well locations remaining on the plate 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 e Press the CLEAR key to clear values on the display e Press ENTER to save the displayed value and advance to the next screen ELx808 Operator s Manual 3 29 NUMBER OF SAMPLE REPLICATES Use this option to enter the number of replicates per sample that
28. OPEN PLATE ACCESS DOOR This command opens the robotic plate access door B 20 Computer Control host t valid limits no arguments CLOSE PLATE ACCESS DOOR This command closes the robotic plate access door host y valid limits no arguments GET INSTRUMENT CONFIGURATION 51 This command returns a 16 bit binary encoded word defining the instrument configuration host 2 valid limits no arguments Instrument configuration data is returned low byte first code 0x0000 0x0001 0x0008 0x0009 0x0080 0x0081 0x0088 0x0089 Appendix B description ELx808 UV ELx808 ELx808 UV w Incubator ELx808 w Incubator ELx808 UV w Robot Access Door ELx808 w Robot Access Door ELx808 UV w Incubator and Robotic Access Door ELx808 w Incubator and Robotic Access Door response lt ACK NAK gt status string 5 response lt ACK NAK gt status string 5 response lt ACK NAK gt configuration 2 no status string Encoding is defined as follows B 21 4 CHECK FEATURE 2 This command answers queries regarding features which may have been added after the initial release of an instrument host valid limits Zi no arguments data bytes 1 4 updated feature 1001 well set reads 1002 reads using two wavelength sets 1003 area scanning 1004 serial wavelength configuration 1005 shaking 1006 absorbance reads
29. SMP SMP SMP SMP SMP SMP SMP SMP SMP C STD2 SMP SMP SMP SMP SMP SMP SMP SMP SMP D STD2 SMP SMP SMP SMP SMP SMP SMP SMP SMP E PC SMP SMP SMP SMP SMP SMP SMP SMP SMP F PC SMP SMP SMP SMP SMP SMP SMP SMP G NC SMP SMP SMP SMP SMP SMP SMP SMP H NC SMP SMP SMP SMP SMP SMP SMP SMP Appendix D Figure D 6 Assay Detail Report D 8 Assay List Version 2 82 1 _Quick Read 2 ASSAY 02 3 ASSAY 03 4 ASSAY 04 5 ASSAY 05 6 ASSAY 06 7 ASSAY 07 8 ASSAY 08 9 ASSAY 09 10 ASSAY 10 11 ASSAY 11 12 ASSAY 12 13 ASSAY 13 14 ASSAY 14 15 ASSAY 15 16 ASSAY 16 17 ASSAY 17 18 ASSAY 18 19 ASSAY 19 20 ASSAY 20 21 ASSAY 21 22 ASSAY 22 23 ASSAY 23 24 ASSAY 24 25 ASSAY 25 26 ASSAY 26 27 ASSAY 27 28 ASSAY 28 29 ASSAY 29 30 ASSAY 30 31 ASSAY 31 32 ASSAY 32 33 ASSAY 33 34 ASSAY 34 35 ASSAY 35 36 ASSAY 36 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 55 ASSAY 55 Figure D 7 Assay List Report Format 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 trans formation calculation and a POS NEG cutoff determination and another with a standard curve possible to translate the kit
30. Test Plate wells C01 D04 E02 F05 G03 and H06 compared to known standard values contained in the Test Plate Data Sheet If the test fails Examine the glass filters on the test plate to see if they are dirty If needed clean them with lens paper Note Do not remove the filters from the test plate and do not use alcohol or other cleaning agents to clean them gt Check the filter calibration values entered in the ELx808 software to make sure they match the values on the Test Plate Data Sheet gt Check the Calibration Due Date on the Test Plate if it has expired the plate must be re calibrated Contact Bio Tek Repeatability This test ensures that the reader meets its repeatability specification by reading each Test Plate 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 instrument If the test fails gt Examine the glass filters on the test plate to see if there are any loose particles that may have shifted between readings and caused changes If needed clean the filters with lens paper Note Do not remove the filters from the test plate and do not use alcohol or other cleaning agents to clean them gt Check the microplate carrier to ensure that it is clear of debris Regarding Linearity Linearity of the Optical Density readings is confirmed by default 1f the readings are accu
31. Use the RIGHT ARROW and LEFT ARROW keys to move to the next or previous editable field e Press the CLEAR key to clear the Standard Concentration value from the display e Press the ENTER key to save the value on the display and move to the next screen ELx808 Operator s Manual 3 25 REUSE OF STANDARD CURVES The ELx808 has the ability to reuse a standard curve that has already been established Limitations of the Reuse of Standard Curves e The reuse of standard curves can only be done in assay positions 31 through 55 Each of these positions can only store one standard curve e Standard curves cannot be reused on panels see page 3 47 for Panel Definition e Standard curves will be stored in memory with the Assay Name Standard Concentrations Replicate Counts and Optical Densities for each standard replicate e Stored standard curves can only be reused for the assay that the curve was originally generated on i e the curve for assay 53 cannot be applied to samples on a plate to be run in assay 51 e To reuse a standard curve an assay must first be programmed in positions 31 through 55 and run 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 has been added after these prompts REUSE STANDARD CURVE YES O e After the assay has been run the results have been calculated and the reports have
32. are assigned to sample wells as follows gt EQUIV if the sample equals 1 500 gt POS if the sample is greater than 1 500 gt NEG ifthe sample is less than 1 500 2 Fora quantitative assay samples with OD values greater than the STD2 mean plus a 10 greyzone should be labeled as positive samples with OD values less than the STD2 mean minus the 10 greyzone should be labeled as negative All other samples should be considered equivocal e For this example STD2 x the mean of the STD2 wells equals 2 000 OD e The cutoff formula is simply STD2 x e The greyzone is 10 e POS is selected for SAMP gt CUTOFF 10 e Calls are assigned to sample wells as follows gt 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 ifthe sample is less than 1 800 ELx808 Operator s Manual 3 39 3 40 TRANSFORMATION FORMULAS Transformation formulas change the absorbance data of all wells defined in the Map to another format in preparation for further data reduction TRANSFORMATION FORMULA DEFINITION e From the assay Define Menu press the arrow corresponding to Formula FORMULA CURVE e This will bring you to a screen asking to Select Formula Type At this screen select TRANS and then enter the formula using the Math Other Map and Function keys ECT FORMULA TYPE OF F TRANS A L Example Divide all ODs on the plate by 2 and m
33. been generated the reader will prompt if this standard curve should be stored in memory The following display will appear SAVE STANDAROD CURVE Y E O e 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 3 26 Operation STANDARDS O N e 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 page 2 15 for more information on the UTIL options e 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 samples 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 standards curves will be routinely reused NUMBER OF CONTROLS The Number of Controls screen allows you to enter the number of controls that will be used in the assay Any previously defined value will appear on the display ENTER NUMBER O F CONTROLS e Use
34. care 60 61 kinetic interval seconds 0 9999 62 63 kinetic read count 2 9999 64 66 don t care 67 69 first wavelength 000 340 900 70 84 remaining wavelengths 000 340 900 85 162 don t care Appendix B B 15 163 164 shake time seconds 0 999 165 shake speed 0 slow 1 med 2 fast 3 variable 166 167 total kinetic read time minutes 1 9999 168 delay before first read 0 no 1 yes 169 170 delay seconds 0 999 status string 5 The multi wavelength mode returns a plate of data for each wavelength specified To select the multi wavelength mode the measurement wavelength must be set to 000 and wavelengths specified starting with first wavelength Up to three wavelengths may be specified in this mode However if dual wavelength subtraction is desired the measurement and reference wavelengths must be specified A single wavelength read may use either mode If a wavelength is not to be used it must be replaced by 000 If the multi wavelength mode is selected the ELx format for data response will be used until a new assay definition table is specified A kinetic interval of zero will cause the plate to be read repeatedly with no pauses between reads The assay definition table must be downloaded before other plate specific commands are sent such as Set Row Range and Set Column Range To minimize confusion all don t cares sho
35. eel eie radere ee in peine eec redi esl 3 43 Edit Standard Outliers ae erae eeoa aar aoa karper nc nn nono eara AO EEE E eNA OAA ESEESE Eni 3 45 Axis Selectlon ce eee eerte eir ete dt is 3 46 Extrapolation of UNKNOWNS ooooocococcccnonoccnonononcnnnnnnccnnno nn eene eene ennemis 3 46 Panel ASSAYS M 3 47 Reading a Microplate nice nan aee Ta a eE a ANa a AAEE A TR EAEAN ATRE 3 50 Select ASSAY E E ees bee E 3 51 Run Time Prompts 3 e aii 3 51 Enter Number of Samples a a e e 3 52 Enter Plate 1D 000 iaa 3 52 Enter Sample ID 5 2 1 tide EEUU ei ipn 3 53 Prompts for Well Locatiori 1 iei tii dete ette ex ee bo t EDUC dee e Le een 3 53 Beginning the Plate Read toti tette ede ip cete et pP Decide 3 53 Printing Reports and Assay Lists ooooooocccnnnncccnnnoccconoconnnonorn cc nano nnnnnnnn enm eene ennemis 3 54 Resulta 3 55 Editing Standard Outliers 5 ii EX dene 3 55 Printing Results 1 cione tied A Ee nieder deci 3 56 MD deret bet t RU ER d rb EE 3 57 CC EU 3 57 Nr 3 57 Chapter 4 Performance Verification and IQ PQ OQ Tests 4 1 Recommendations for Achieving Optimum Performance seen 4 2 Installation Qualification IQ riter a 4 3 Recommended Test and Maintenance Schedule OQ PQ PM see 4 4
36. entry module and turn it over Changing to the e Remove the two 630 mA fuses pears Ys e Install one AG 1 5 A 1 5 A 250 V fuse PN 46024 on the fuse onfiguration holder on the side opposite the original 630 mA fuse location e Reinstall the fuse holder with the fuse toward the back of the power entry module e Replace the cover and verify that the white plastic indicator pin lines up with the correct power entry module indicator hole Changing to the e Remove the single AG 1 5 A fuse MNA LAM e Install two 0 63 A 630 mA 250 V fuses PN 46038 on the fuse onliguration holder on the side opposite the original AG 1 5 A fuse location e Reinstall the fuse holder with the fuses toward the back of the power entry module e Replace the cover and verify that the white plastic indicator pin lines up with the correct power entry module indicator hole 2 8 Installation To replace defective fuses 1 Follow steps 1 3 on the previous page to access the fuse holder 2 Remove the defective fuse and replace it with the correct new one 3 Reinstall the fuse holder with the fuse toward the back of the power entry module 4 Replace the cover and verify that the white plastic indicator pin lines up with the correct power entry module indicator hole ELx808 Operator s Manual 2 9 Connect Power 2 10 1 Locate the power inlet on the right side of the ELx808TM 2 Plug the rounded end of the power cord into t
37. formulation is to create a solution that absorbs light at 2 000 OD full strength when dispensed at 200 pl well in a flat bottom microplate Alternatively any solution that gives a stable color will suffice This includes substrates incubated with an enzyme preparation and then stopped with an acidic or basic solution Some enzyme substrate combinations that may be used as alternates to the described dye are shown in the table below Table 4 2 Typical Enzyme Substrate Combinations and Stopping Solutions Alkaline Phosphate o nitrophenyl phosphate 3N sodium hydroxide beta Galactosidase o nitrophenyl beta D galactopyranoside 1M sodium carbonate Peroxidase Peroxidase 2 2 Azino di ethylbenzothiazoline sulfonic citrate phosphate buffer pH 2 8 acid ABTS o phenylenediamine 0 03N sulfuric acid ELx808 Operator s Manual 4 17 Stock Solution Formulation The stock solution for Liquid Test 1 and 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 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 Deionized water e Precision balance with readability of 0 001 g e Weigh boat e 1 liter volumetric flask Table 4 3 Stock Solution Formulation for Liquid Tests 1 and 2 FD amp C Yellow No 5 powder 0 092
38. in the filter wheel To check and or adjust the filter table e Turn on the reader if it is not already on At the Main Menu screen shown below press the soft key under the UTIL menu option M 10 09 04 EPORT UTIL e The Select Utility Option menu appears SELECIT UTILIT TESTS SETUP O e From the Select Utility Option menu press the soft key beneath SETUP The Edit Setup Information screen appears EDIT SETUP INFORMATION DATE TIME FILTER MOR E e From this menu press the soft key beneath FILTER The wavelength for Filter 1 appears ENTER FILTER 1 WAVELENGTH e Ifyou need to change the filter wavelength number use the NUMERIC keypad to enter a number at the cursor location To save the entry and move to the next filter on the filter table press the ENTER key e When the last filter has been entered the software exits the filter routine and displays the following screen EDIT SETUP INFORMATION DATE TIME FILTER MOR E e Press the MAIN MENU key to return to the Main Menu 2 14 Installation Configure Utility Options The ELx808 may be configured a number of ways depending on user preference Configuration options are accessed via the Utility Options menu and include e SETUP Set the date and time e OUTPUT Select whether reports will be output to a Printer the Computer or both choose the Column or Matrix report format and determine if Curves will be printed e READ Indic
39. 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 beep 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 Panels Map Report to verify the map before reading the plate Choose Report from the Main Menu Map and 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 ELx808 Operator s Manual 3 49 Reading a Microplate Press the READ option found at the Main Menu to read a microplate e From the MAIN MENU screen press the soft key beneath the READ menu option to access the SELECT ASSAY NUMBER screen e Alternately press the red READ key on the lower right of the keyboard SELECT ASSAY At the Select Assay Number screens e Use the NUMERIC keys to enter th
40. 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 Choose to blank on AIR if no blank BLANK MAP AIR wells are required Appendix E E 5 STEP NUMBER STDS 00 REUSE STANDARD CURVE NUMBER CTLS 03 CONTROL 1 PC CONTROL 2 CTL1 CONTROL 3 NC NUMBER OF REPLICATES PC 01 CTL1 03 NC 01 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 NC x 0 300 NO OF REPLICATES 01 E 6 COMMENTS Can only be programmed in assay positions 31 55 and cannot be reused on panels Suggested choice for the cutoff control 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 Instructions for Programming a New Assay STEP COMMENTS 3 To create the plate transformation for CALC divide the ANA result by a cutoff standard FORMULA MORE TRANS VAR SCOPE VARIABLE SMP or OD Select OD to advance to the formula definition screen and define the trans
41. of results from the plate reading The form in which the results are presented is determined by the report settings Matrix Column Curve Fit specified in the UTILITIES menu option e Select MAP to print a matrix showing the locations of the Blanks Standards Controls and Samples for a selected 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 ELx808 Note The reader stores measurement values for the last 8 plates in memory 3 54 Operation Result 07 17 09 5 e Use the OPTION key to select the appropriate Plate ID and Report Note that the Assay ID will change if the selected Plate ID was read with a different assay Once you have found the correct Plate ID press ENTER 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 EDIT STD 1 RE P 1 YES
42. on cuv ul s Map Direction ACROSS Rep Direction DOWN p 0 AL A quc A sso pes Ee aa STDI STD2 STD3 STD4 STD5 rc NC SMP sme STDI STD2 STD3 STD4 STD5 PC NC SMP SMP Lo p Lo LLL m npn E A ie t Ep ees Lo po p lll y Eq rii Lx lw Ea ru A AE A R MES D ELx808 Operator s Manual 3 19 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 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 NUMERIC and ALPHA keys to enter a letter or number at the cursor location For all prompts of a well location only the ALPHA keys are active for the first character and NUMERIC for the second and third characters e Press ENTER to save the well location and advance to the next screen SELECTING A BLANK MAP This option allows you to select which blanking method to apply to the assay The currently selected Blank Map value appears on the top line of the display and the available options appear on the bottom The blanking options AIR FULL and CONSTANT ROW and COLUMN and P ACROSS and P DOWN are displayed on three screens 3 20 Operation e Press SOFT
43. pursuant to Part 15 of the FCC Rules These limits are designed to provide reasonable protection against harmful interference when the equipment is operated in a commercial environment Like all similar equipment this equipment generates uses and can radiate radio frequency energy and if not installed and used in accordance with the instruction manual may cause harmful interference to radio communications Operation of this equipment in a residential area is likely to cause interference in which case the user will be required to correct the interference at his own expense Canadian Department of Communications Class A This digital apparatus does not exceed Class A limits for radio emissions from digital apparatus set out in the Radio Interference Regulations of the Canadian Department of Communications Le present appareil numerique n met pas du bruits radioelectriques depassant les limites applicables aux appareils numerique de la Class A prescrites dans le Reglement sur le brouillage radioelectrique edicte par le ministere des Communications du Canada Safety This device has been type tested by an independent laboratory and found to meet the requirements of e Underwriters Laboratories Standard UL UL3101 1 Electrical Equipment for Laboratory Use Part 1 General Requirements e Canadian Standards Association CAN CSA C22 2 No 1010 1 1992 Safety Requirements for Electrical Equipment for Measurement Control and Laboratory Use Part
44. rear quadrant inside the reader Remove the filter wheel by pulling it off its magnetic hub 3 Inspect the glass filters for speckled surfaces or a halo effect This may indicate deterioration due to moisture exposure over a long period of time If you have any concerns about the quality of the filters contact your Bio Tek representative 4 Clean the filters using lens cleaning tissue moistened with a small amount of isopropyl ethyl or methyl alcohol Ensure that the filters remain in their current locations 5 Replace the filter wheel on its magnetic hub Ensure that the peg on the wheel lines up with the notch on the hub and that the filter wheel is positioned flat against the hub and rotates freely 6 Close the top shroud and replace the seven perimeter screws Notes e Store filters in a dry environment such as a desiccator if they will not be used for an extended period e If the reader continues to fail tests in this chapter after you have cleaned the filters and bulb contacts or if one or more filters appear to have deteriorated contact your Bio Tek Representative or Bio Tek ELx808 Operator s Manual 4 29 Periodic Maintenance for ELx808 Robotic Units 1 Remove the top shroud of the instrument see instructions in Chapter 2 2 Lubricate the components as shown in Figures 4 4 and 4 5 every six months or after 10 000 cycles according to the instructions in the table below Note Use Lubricant Bio Tek PN 66039
45. rising out of breach of Warranty or otherwise The implied warranties of merchantability and of fitness for any particular purpose are expressly excluded and do not apply to the instrument software This warranty shall not be changed or modified in any way without the express written permission of an officer of Bio Tek Instruments Hardware Warranty Bio Tek warrants the hardware instrument for a period of one 1 year from the date of the product shipment from Bio Tek against defective materials and workmanship This warranty is limited to the original purchaser and cannot be assigned or transferred All claims under this limited warranty must be made in writing to Bio Tek Attention Service Department or to Bio Tek s authorized representative Purchaser must ship the product to Bio Tek postage pre paid Bio Tek shall either repair or replace with new like new or equipment of similar condition at its option and without cost to the purchaser any product that in Bio Tek s sole judgment is defective by reason of defects in the material or workmanship This warranty is void if the instrument 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 1s void if the instrument has been repaired or altered by persons not authorized by Bio T
46. runs an internal System Test each time it is powered on The reader will beep 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 via the keypad KC4 or KCjunior to try to retrieve an error code from the reader To obtain a printout of the System Test values for either periodic testing and documentation or troubleshooting select SYSTEM from the TESTS submenu The results are sent to the printer via the reader s parallel port and a printout in pass fail format similar to Figure 4 1 is produced falling below 30 000 for filter wavelengths of 405 nm or higher Liquid Test 1 should be performed This test is applicable to the eight reading channels only not the reference channel D Note If a System Test shows one or more channels that have air readings Note The incubation portion of the System Test is only performed on incubation equipped instruments T version ELx808 Operator s Manual 4 7 01 14PM 01 01 04 SYSTEM SELF TEST Filter 405 Gain 10 67 Channel Ref 1 2 3 4 5 6 7 8 Air 26296 51089 46917 52883 46941 45790 50010 48537 42439 Dark 1555 1789 1796 1507 1576 1590 1403 1637 1600 Delta 24741 49300 45121 51376 45365 44200 48607 46900 40839 Filter 450 Gain 3 82 Channel Ref 1 2 3 4 5 6 7 8 Air 26967 51694 49258 54304 48807 46335 51879 49654 45529 Dark
47. t match original passed in B 26 Computer Control Appendix C Error Codes This appendix provides a list of Error Codes that may appear on the ELx808 and their probable causes If an error code appears on the display or in KCjunior K C4 look up the code in the Troubleshooting tables in this appendix and attempt to solve the problem If you are unable to solve the problem contact Bio Tek s Technical Assistance Center See page 1 7 for contact information Glossary of Terms gt Air Blank A full light reading through a filter with no plate in the light path DarkCurrent 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 gt 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 Axis Refers to a motor that positions the filter wheel or plate carrier gt Offset A numerical limit usually a range For example if the gain fails an offset test it may be too high or too low Error Codes Error Codes Displayed Error Potential Cause ERROR 0200 Plate carrier did not find the home sensor ERROR 0201 Filter wheel motor did not find the home sensor filter wheel not installed or detached from hu
48. the NUMERIC keys to enter the Number of Controls The range depends on the number of locations on the plate that are undefined The maximum number of controls is 8 e Press CLEAR to clear the value on the display e Press ENTER to save the value and advance to the next screen ELx808 Operator s Manual 3 27 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 CONTROL 1 PC Cc e Press the soft keys under the displayed Control Type to select the option Positive Control Negative Control High Positive Control Low Positive Control CTL1 CTL2 CTL3 CTLA e Press CLEAR to clear the Control Type from the display e Press ENTER to save the displayed Control Type and advance to the next screen NUMBER OF CONTROL REPLICATES The Number of Control Replicates screen allows you to enter the number of replicates per control group in the assay Any previously defined number will be displayed ENTER NUMBER O REPLICATES O F e Use the NUMERIC keys to enter a value for Number of Control Replicates The range is 1 to 12 replicates The software performs a check to ensure the number of replicates does not exceed the number of undefined wells remaining on the plate e Press CLEAR to clear the displayed Number of Replicates value e Press ENTER to save the displayed value and advance to the next screen 3 28 Operation LOCATION
49. which will calculate the time for each well to reach the onset optical density e Press ENTER to save the selection and advance NUMBER OF KINETIC POINTS SELECTION Use this screen to select the number of sequential kinetic points to calculate the steepest Rate or the R squared at the steepest Rate KINETIC POINTS ALL POINTS e Use the NUMERIC keys to input the number of points The range is 2 to MAX where max 1s the total number of reads e Press SOFT KEY 1 or 2 to select ALL POINTS e Press ENTER to save the entry and advance to the next screen 3 14 Operation ONSET OD SELECTION Use this screen to enter the onset OD time e Use NUMERIC keys to enter the onset OD 3 000 OD is the maximum value e Use the LEFT and RIGHT ARROW keys to move the cursor within the entered OD field e Press the ENTER key to save the entry and advance LINEAR SCANNING If Scanning is chosen as the Read Type use the following screen to enter the total number of points to be read in a line across the center of each well ENTER NUMBER O F SCAN POINTS The maximum number of points selectable is 31 odd numbers only The 31 scan positions are fixed in the software You must determine the optimal number of scans per well If for example 7 scans across the well is chosen the reader will read the centermost seven points in the well The more scan points chosen the closer to the well sides reads will be taken Note If to
50. will be run in the assay Any previously defined value will be displayed ENTER NUMBER O F SAMPLE REPLICATES e Use the NUMERIC keys to enter the Number of Sample Replicates The range is 1 to 12 replicates The software ensures that the number of replicates multiplied by the number of samples multiplied again by the number of dilutions does not exceed the number of undefined wells remaining on the plate e Press the CLEAR key to clear the value on the display e Press the ENTER key to save the displayed value and advance to the next screen SAMPLE LOCATION If Manual Map Generation is selected this screen allows you to select the well location of the sample on the plate Any previously defined Sample Location appears on the display SAMPLE 1 LOCATION RE P 1 A0 1 e Use the NUMERIC ALPHA and DECIMAL POINT keys to enter the sample identifier and its location on the plate e Press the CLEAR key to clear the displayed value e Press ENTER to save the value and advance to the next screen 3 30 Operation FORMULA Defining the Formula involves identifying the formula type and entering the actual formulas Formula definition screens are displayed on the ELx808 in the order detailed below Formulas created using Bio Tek s Extensions Define Protocol software cannot be edited by using the reader Formulas are processed in the following order with the number of permitted formulas of each type Blank Valida
51. 01 0 002 PASS C12 0 002 PASS H08 0 003 PASS Al Top Left Accuracy Results col D04 E02 FO5 G03 HO6 STANDARD 0 156 2 812 0 620 2 264 1 192 1 754 DATA 0 147 2 824 0 618 2 276 1 197 1 757 RESULT PASS PASS PASS PASS PASS Repeatability Results col D04 E02 F05 G03 H06 READ 1 0 147 2 824 0 618 2 276 1 197 1 757 READ 2 0 147 2 817 0 618 2 275 1 197 1 757 RESULT PASS PASS PASS PASS PASS H12 Top Left Accuracy Results col D04 E02 F05 G03 HO6 STANDARD 0 156 2 812 0 620 2 264 1 192 1 754 DATA 0 146 2 812 0 618 2 274 1 196 1 756 RESULT PASS PASS PASS PASS PASS Repeatability Results col D04 E02 F05 G03 HO6 READ 1 0 146 2 812 0 618 2 274 1 196 1 756 READ 2 0 146 2 815 0 618 2 275 1 196 1 756 RESULT PASS PASS PASS PASS PASS Figure 4 3 Sample Absorbance Plate Analysis Report ELx808 Operator s Manual 4 13 4 14 The Calibration Report has three categories for full test descriptions refer to page 4 10 Alignment This portion of the test measures the alignment of the microplate carrier with the optical path A reading greater than 0 015 OD for alignment wells F01 C12 or H08 represents an out of alignment condition If the test fails Examine the microplate carrier to ensure that it is clear of debris Make sure the Test Plate is properly seated in the microplate carrier gt Check the Test Plate s alignment holes to ensure they are clear of debris Accuracy Accuracy is a measure of the absorbance optical density of
52. 5 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 0 5 0 005 OD from 0 to 2 500 OD 0405 nm CHANNEL TO CHANNEL VARIATION AND ALIGNMENT TEST C 4 24 l 3 Using the plate prepared for Test A on the previous page conduct a Turnaround test by reading the plate with the A1 well in the H12 position five times This test results in 2 comparisons of each channel to its corresponding channel Calculate the means of the wells in column 1 in the Normal plate position data is from Test A and in the turnaround position from Step 1 above Compare the mean reading for well A1 to its mean reading when in the H12 position Next compare the mean values for the other wells to their corresponding mean values with the well in the turnaround position Compare B1 to G12 C1 to F12 D1 to E12 El to D12 F1 to C12 Gl to B12 H1 to A12 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 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 0 019 0 010 0 029 which is added and subtracted from 1 902 for the range If any set of well values is ou
53. 6 ERROR 1000 ERROR 1100 ERROR 1200 ERROR 1300 ERROR 1400 ERROR 1500 ERROR 1600 ERROR 1700 ERROR 1800 ERROR 1900 ERROR 2311 2386 Potential Cause Memory Manager corruption detected Power instrument off then back on Reader failed calibration checksum test Internal software corruption Wavelength not detected in reader s filter table Compare filters wavelengths to the filter table Reader s filter or channel signal is out of specified range Check for lamp failure and or replace lamp Required reader configuration data missing Re download software basecode and assays Failed configuration checksum test Re download software basecode and assays Calibration data missing Motor not homed Autocal error see error 0200 Assay incubation Error Assay requires incubation but the instrument does not appear to have an incubator Incubator failed to hold temperature within tolerances during the assay Computer control assay definition error This error is used only for software development Kinetic interval too short for selected options Too many kinetic intervals selected Memory allocation failure This error is used only for software development Lamp unstable Clean lamp contacts page 4 28 Error Codes Appendix D Report Format The following are examples of reports that can be generated and or printed from the ELx808 M For details on how to print these reports refer to Output O
54. Dev CV Notes p M H A A E A01 STDI 0 182 0 182 0 200 B01 STD2 0 529 0 529 0 400 0 483 C01 STD3 0 721 0 721 0 600 0 686 001 STD4 0 749 0 749 0 800 0 715 E01 SMPOQOI 0 068 0 068 POS F01 SMP0002 0 083 0 083 POS 601 SMP0003 0 073 0 073 POS H0 SMPO004 0 072 0 072 POS A02 SMPO005 0 072 0 072 POS 802 SMP0006 0 076 0 076 POS C02 SMPO007 0 070 0 070 POS 002 SMPO008 0 081 0 081 POS 02 SMPO009 0 070 0 070 POS F02 SMPO010 0 069 0 069 POS 602 SMPO011 0 070 0 070 POS H02 SMPO012 0 075 0 075 POS A03 SMPO013 0 073 0 073 POS Page 1 Figure D 3 Samples with Calls on Column Report Report Format Appendix D Bio Tek Instruments Assay ASSAY 01 Date 07 29 96 Lot Time 09 55 19AM Operator Wavelength 405 Temp Plate ID pa aaaaaaaaaassssssssasasasststststststststst t t t t t t t l l l ltlutlullli iaiaiiMMMlMl COMMENTS OUT indicates an out of range value was read Interpretation of Results GREYZONE 0 25 0 250 CUTOFF 1 0 100 SMP SMP lt CUTOFF CUTOFFFGREVZONE AND SMP gt CUTOFF CUTOFFSGREYZONE EQUIV SMP SMP gt CUTOFF CUTOFF GREYZONE NEG SMP SMP CUTOFF CUTOFFKGREYZONE POS LL i a AAA K M Well ID B1kd0D Calco Call Pred Conc RSLT Std Dev C Notes O AA AD c A01 STD 0 184 0 184 B01 STD 0 529 0 529 cor PC 0 723 0 723 D NC 0 746 0 746
55. ELx808 Absorbance Microplate Reader Operator s Manual ANN f A ELx808 for all ELx808 models BIO TEK Creating Solutions for the Future of Science 7341000 Rev K1 July 2005 This Manual Update contains a change to the ELx808 Operator s Manual This change will be incorporated in the next full revision Rev L of the Manual Page 4 20 Liquid Test 1 Recommendation After pipetting the diluted test solution into the microplate step 3 and before reading the plate we strongly recommend shaking the plate at Variable speed for four minutes This will allow any air bubbles in the solution to settle and the meniscus to stabilize Alternatively wait 20 minutes after pipetting the diluted test solution before reading the plate GJSBIO TEK BIO TEK INSTRUMENTS INC P O BOX 998 HIGHLAND PARK WINOOSKI VERMONT 05404 USA COPYRIGHT O 2005 TEL 888 451 5171 FAX 802 655 7941 Outside the USA 802 655 4740 E mail customercare biotek com www biotek com Creating Solutions for the Future of Science ELx808 Absorbance Microplate Reader Operator s Manual APRIL 2005 O 2005 PART NUMBER 7341000 REVISION K BIO TEK INSTRUMENTS INC Preface Notices BIO TEK INSTRUMENTS INC P O Box 998 Highland Park Winooski Vermont 05404 0998 USA Customer Service and Sales Internet www biotek com Phone 888 451 5171 toll free in the U
56. ER OF SAMPLES Use this screen to enter from 00 to the maximum number of samples permitted by the previously created well map If there are no controls standards or blanks defined the minimum number of samples is 1 This value controls the number of samples reported if Matrix or Column reports are requested ENTER NUMBER O F SAMPLES ENTER PLATE ID You can enter a 10 character maximum identifier to assign to the plate Since this Plate ID will be stored in the reader s memory each plate ID should be unique Note Use caution when creating multiple Plate IDs The reader does not warn you that you are about to exceed the maximum of 10 plate IDs stored in memory If an eleventh Plate ID is added it will overwrite the first Plate ID stored in memory Note Ifthe internal bar code scanner option is installed the reader will automatically scan the plate bar code label and use this as the Plate ID e Use the KEYPAD to enter numbers and the LEFT RIGHT arrow keys to move the cursor CLEAR clears the display 3 52 Operation ENTER SAMPLE ID Y ou can start entering a starting sample identification number from 0001 to 9999 The software will automatically increment each subsequent sample identification number by 1 Sample IDs will be assigned according to the previously defined mapping order e Use the KEYPAD to enter numbers and the LEFT RIGHT arrow keys to move the cursor CLEAR clears the display PRO
57. End of Report Figure D 4 Column Report without Samples D 5 D 6 Bio Tek Instruments Panel Date 07 31 96 Lot Time 02 24 01PM Operator Temp Plate ID COMMENTS Specimen Assay Well B1kd0D Calc0D Call RSLT Std Dev V4 Notes SMP0001 ASSAY 01 F0 006 006 ASSAY 02 B05 0 000 0 000 SMP0002 ASSAY 01 601 0 002 0 002 ASSAY 02 C08 0 007 0 007 SMP0003 ASSAY 01 HOY 0 005 0 005 ASSAY 02 005 0 004 0 004 SMP0004 ASSAY 01 A02 0 002 0 002 ASSAY 02 E05 0 012 0 012 SMP0005 ASSAY 01 B02 002 002 ASSAY 02 FOS 0 010 0 010 SMP0006 ASSAY 01 C02 0 005 0 005 ASSAY 02 605 001 001 SMP0007 ASSAY 01 002 001 001 ASSAY 02 HOS 0 002 0 002 SMP0008 ASSAY 01 E02 0 009 0 009 ASSAY 02 A06 0 004 0 004 SMP0009 ASSAY 01 F02 0 006 0 006 ASSAY 02 B06 0 007 0 007 SMP0010 ASSAY 01 602 006 006 ASSAY 2 C06 0 010 0 010 SMP0011 ASSAY 01 H02 005 005 ASSAY 02 DOG 0 004 0 004 SWP0012 ASSAY 01 A03 00 00 ASSAY Q2 E06 0 003 0 003 SMP0013 ASSAY 01 B03 0 002 0 002 ASSAY 02 F06 001 001 SMP0014 ASSAY 01 003 005 005 ASSAY 02 G06 0 001 0 001 SMP0015 ASSAY 01 D03 002 002 Page Figure D 5 Panel Report Report Format 03 34PM 11 11 03 NAME ASSAY 55 READ TYPE ENDPOINT WAVELENGTH 450 BLANK MAP AIR AVERAGE STANDARDS NO CONCN STD1 6 2 STDZ 12 5 CURVE FIT TYPE LINEAR EDIT STD OUTLIERS NONE X Y AXIS TYPE LIN EXTRAPOLATE UNKNOWNS NO 1 2 3 4 5 6 7 8 9 10 11 12 A STDi SMP SMP SMP SMP SMP SMP SMP SMP SMP B STD1
58. IN MENU key Running the Absorbance Plate Test From the reader s Main Menu select UTIL gt TESTS gt CALPLTE CALPLT amp E CALIBRAT 40 5 4 e The test is run using one filter wavelength Select the desired wavelength from the CALIBRATION FILTER screen e When prompted insert the Test Plate into the ELx808 s plate carrier and press the READ key to begin the calibration program e The plate will be read twice and the reader will prompt you to rotate the plate 180 Rotate the plate so that H12 is in the Al position and then press the READ key to complete the test e The Calibration Plate Analysis Report Figure 4 3 on the next page will print when the test is complete 4 12 Performance Verification and IQ OQ PQ Tests Important The ELx808 s Absorbance Plate Test tests the accuracy and repeatability specifications from 0 000 to 2 500 OD only The Calibration Plate Analysis report displays the OD value read in well position D04 in this example but does not indicate PASS or FAIL because the value 1s higher than 2 500 OD and therefore is not within the software test range Important The Absorbance Plate Test and CALPLATE utility in the reader are used only to verify the reader s performance The Test Plate and utility DO NOT correct or calibrate the instrument CALIBRATION PLATE ANALYSIS 08 51AM 06 21 02 Read Mode Normal Filter 405 Operator ID Notes Al Top Left Alignment Results F
59. If only one of the four mean values results in a failure check the well for debris and the plate for scratches or fingerprints ACCURACY SPECIFICATION For comparison in this test the following accuracy specification is applied using Normal mode and a 96 well microplate 1 0 0 010 OD from 0 to 2 500 OD 2405 nm ELx808 Operator s Manual 4 21 Liquid Test 2 Note A 96 well flat bottom microplate is required for this test Corning Costar 3590 is recommended Use a new microplate any fingerprints or scratches may cause variations in the turnaround reading REQUIRED MATERIALS e A new 96 well flat bottom microplate 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 see page 4 18 PREPARATION OF DILUTIONS Create a percentage dilution series beginning with 100 of the original concentrated stock solution A or B in the first tube 90 of the original solution in the second tube 80 in the third tube all the way to 10 in the last tube Dilute using amounts of the remaining 0 05 solution of deionized water and Tween 20 as shown below Table 4 4 Test Tube Dilutions Tube Number 1 2 3 4 5 6 7 8 9 10 Volume of Original 20 18 16 14 12 10 4 2 Solution ml Volume of 0 05 2 4 10 12 14 16 18 Tween Solution ml Absorbance expected i
60. MAP AUTO DOWN DOWN A01 BLANK MAP AIR NUMBER STDS 05 NUMBER STD REPLICATES 01 CONCENTRATIONS STDI 6 25 STD2 12 5 STD3 25 STD4 50 STD5 100 COMMENTS Instructions for Programming a New Assay STEP REUSE STANDARD CURVE NUMBER CTLS 02 CONTROL 1 PC CONTROL 2 NC NUMBER OF REPLICATES PC 01 NC 01 SAMPLES 89 SAMPLE REPLICATES 01 To define a cutoff formula for Positive and Negative calls 4 Curve 4P Appendix E COMMENTS Kit instructions specify that samples with concentration values greater than 11 should appear as positive The ELx808 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 Instructions for Programming a New Assay Appendix F Instructions for Adjusting the Line Input Voltage Range This appendix provides instructions for adjusting the line input voltage range and replacing fuses for ELx808 instruments manufactured with an earlier version of the power entry module than described in Chapter 2 pag
61. MP17 SMP25 SHP33 SMP41 SHP49 SMPS7 SHP65 SMP73 SMP81 SMP89 Calc0D 0 073 0 070 0 073 0 072 0 076 0 072 0 074 0 080 0 076 0 072 0 072 0 073 Wel SMP3 SMP11 SMP19 SMP27 SMP35 SMP43 SWP51 SMP59 SMP67 SMP75 SNP83 SMP91 CALL POS POS POS POS POS POS POS POS NEG POS POS POS CalcOD 0 072 0 075 0 073 0 074 0 081 0 069 0 076 0 069 0 738 0 076 0 075 0 072 Well SMP4 SWPT2 SMP20 SWP28 SMP36 SMP44 SHP52 SMP60 SMP68 SMP76 SMPB4 SMP92 RSLT 0 704 Figure D 1 Samples with Calls on Matrix Report Report Format Bio Tek Instruments Assay ASSAY 01 Date 07 23 96 Lot Time 03 05 35AM Operator Wavelength 405 Temp Plate ID SONTA COMMENTS OUT indicates an out of range value was read Standard Curve Linear Fit 0 749 P 0 182 ma AA PAR 0 200 0 400 0 600 0 80 CONCENTRATION Curve y Ax 8 Coefficients A 0 946 8 0 073 R Sqr 0 9752 Figure D 2 Curve Fit Report Appendix D D 3 D 4 Bio Tek Instruments Assay ASSAY 01 Date 07 23 96 Lot Time 03 05 35AM Operator Wavelength 405 Temp Plate ID SONTA COMMENTS OUT indicates an out of range value was read Interpretation of Results GREYZONE 0 00 0 000 CUTOFF 2 0 200 SMP SMP lt CUTOFF CUTOFFAGREYZONE AND SMP gt CUTOFF CUTOFF GREYZONE EQUIV SMP SMP gt CUTOFF CUTOFF GREYZONE NEG SMP SMP lt CUTOFF CUTOFF GREYZONE POS mm aaa aaa Well ID B1kd0D CalcOD Call Pred Conc RSLT Std
62. MPTS FOR WELL LOCATION Well locations can be changed at run time if a Manual Map has been specified and you have requested a sample count at run time via the Utilities menu LOCATION RE P 1 aA01 e Use the KEYBOARD to enter the well location using the SHIFT LETTER sequence to key in letters and press ENTER to specify the desired location BEGINNING THE PLATE READ When the following screen appears on the display the reader is ready to read a plate PLACE PLATE IN CARRIER AND PRESS lt READ gt K E Y e Place the plate in the carrier and press the READ key to initiate the plate read After the plate has been read all requested reports will be generated e To halt in read in progress press the STOP key Note If using the incubation option the reader will wait for the incubator to reach temperature before reading the plate ELx808 Operator s Manual 3 53 Printing Reports and Assay Lists Reports are automatically generated after a plate has been read see Output Options and Report Type in Chapter 2 for information on selecting reports To manually regenerate results reports use the REPORT option from the Main Menu You can also print Map Assay and Assay List reports Note See Appendix D for sample reports 9 45PM 05 09 95 DEFINE REPORT UTIL e Press SOFT KEY 3 REPORTS to access additional reports PRINT REPORT RESULT MA P ASSAY LIST e Select the RESULT option to print an exact copy
63. PBS solution use 5 ml as the diluent e fusing the 10x Concentrate PBS Solution mix one part of the concentrate with nine parts of deionized water Then use 5 ml of this solution as the diluent Carefully prepare 50 Test Solution by diluting 10 ml of the 100 Test Solution e fusing the Sigma PBS solution use 10 ml as the diluent e fusing the 10x Concentrate PBS Solution mix one part of the concentrate with nine parts of deionized water Then use 10 ml of this solution as the diluent Pipette the three solutions into the new 96 well microplate e 150 pl of the 100 Test Solution into all wells of columns 1 and 2 e 150 ul ofthe 75 Test Solution into all wells of columns 3 and 4 e 150 ul of the 50 Test Solution into all wells of columns 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 data or export the data to and Excel spreadsheet using KCjunior or KC4TM Performance Verification and IQ OQ PQ Tests REPEATABILITY TEST A 1 For each well calculate the mean and standard deviation of the five readings 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 2 For each mean calculated in step 1 calculate the allowed deviation using the repeatability specification for a 96 well plate of 1 0 0 005
64. S c or d commands host valid limits response BH lt ACK NAK gt data bytes inclusion 0 no temperature with plate data I include temperature with plate data By default the instrument is initialized to not include a temperature response and automatically reverts to this default when a new assay definition table is loaded either from the front panel or by computer control PRESENT PLATE CARRIER J This command causes the plate carrier to move back outside the instrument where it can be loaded with a microplate host valid limits response y no arguments lt ACK gt status string 5 SET READ MODE k This command selects the read mode controlling the speed accuracy and precision of all subsequent plate reads host valid limits response kK no arguments lt ACK gt data bytes mode 0 slow enhanced mode 1 fast mode status string 5 Appendix B B 11 SET STATUS FORMAT n This command selects the format for the status string see Status String Format host valid limits response n no arguments lt ACK gt data bytes format 0 312 status mode 1 ELx status mode status string 5 When the status format is set to ELx status mode any error detected during the execution of any command described in this document will be transmitted to the host PC in this format in addition plate read and scan commands d
65. S 802 655 4740 outside the U S Fax 802 655 7941 E Mail customercare a biotek com Service Technical Assistance Center TAC Phone 800 242 4685 toll free in the U S 802 655 4740 outside the U S Fax 802 655 3399 E Mail tac biotek com European Coordination Center Bio Tek Instruments GmbH Kocherwaldstrasse 34 D 74177 Bad Friedrichshall Germany Phone 49 0 7136 9680 Fax 49 0 7136 968 111 E Mail info biotek de All Rights Reserved 2005 Bio Tek Instruments Incorporated No part of this manual may be reproduced or transmitted in any form or by any means electronic or mechanical including photocopying and recording for any purpose other than the purchaser s personal use without written permission from Bio Tek Instruments Inc Trademarks Bio Tek is a registered trademark and Extensions and Define Protocol are registered trademarks or trademarks of Bio Tek Instruments Inc Microsoft Windows and the Windows logo are registered trademarks or 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
66. 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 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 see Assay Calibration page E 11 Cardiolipin IgG Positive Control 1 bottle containing 1 2 ml of diluted serum The expected value is given on the QC certificate Ready to use Cardiolipin Negative Control 1 bottle containing 1 2 ml of diluted 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 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 Appendix E Automatic Microplate Plate Washer This 1s recommended however plate 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 dispen
67. Started With KC4 0 ccc cccccsceseceeeceseeeeeeseecseecaecesecesecsseeseeeseeeseneeenes B 6 Controlling the Reader Using Serial Protocol sse B 7 Computer Control Command t ooocooconocnconononnnonanannnonnonnccon ccoo coco noco nono nennen B 8 Using the Stop Key to Halt Plate Reads B 23 Status String Formaten oet en ee o eire C be e i id B 24 Overview With the computer control feature the user is provided even more power and flexibility For example the ELx808 can define and run computer controlled kinetic assays using up to three wavelengths on the same microplate When using computer control the ELx808 s onboard blanking and data reduction calculations are suppressed and the raw data is returned to the controlling software for evaluation Readings higher than 3 000 OD may be transmitted This appendix details the protocols necessary for communicating with the ELx808 reader In addition instructions are provided for controlling the reader with Bio Tek s KCjunior or KC4 software packages Computer Control Controlling the Reader With KCjunior Follow these steps to establish communication between the ELx808 and K Cjunior 1 6 7 Appendix B Turn off the computer and the reader Connect the serial cable PN 75053 between the two machines Turn on both machines Install KCjunior on the computer s hard drive Once installed start KCjunior Select Setup then Reader 1 T
68. T KEY 1 to select Control Validation Formula Press SOFT KEY 3 to select Assay Validation Formula Press SOFT KEY 4 to select Blank Validation Formula e Press ENTER to save the validation type and advance ELx808 Operator s Manual 3 33 3 34 FORMULA ENTRY This screen allows you to enter the formula used in the assay Note In formulas OD is used to represent the optical density value FORMUTLA 1 MATH OTHER e After a moment the FORMULA 1 prompt disappears and the formula can be entered e Each formula can contain a maximum of 24 characters Spaces are unnecessary e Use the LEFT and RIGHT ARROW keys to move the cursor to the previous or next editable field e Press SOFT KEY 1 to place the next item on the MATH list at the cursor position The following table shows the order of items on the MATH list pe NNNN C pe Operation reduction A syntax error in a formula will result in a Token Error on D Note The reader software checks the formulas for errors during data results reports e Press SOFT KEY 2 to place the next item on the OTHER list at the cursor position See the table that follows for the order of items on the OTHER list Left parenthesis Right parenthesis AND Logical AND e Press SOFT KEY 3 to place the next defined item on the plate map list 1 e STD NC PC BLK at the cursor position e Press SOFT KEY 4 to place the next option on the F
69. TRANS e Enter OD TVAR 100 3 42 Operation CURVE The CURVE entry screens allow editing and entry of Curve Fit Type Editing of Outliers Axis Identification Extrapolation of Unknowns These screens are displayed on the ELx808 in the order in which they appear in the assay Ifa closed variable 1 e an element of the assay definition that you cannot access or modify is being used in the assay the entry screen is omitted CurveFit The Curve Fit screen allows you to select the curve fit method that will be applied to the assay Any previously defined curve fit type appears on the top line of the display and available options on the second line The Curve Fit screen has three sub menu screens Each sub menu screen provides different curve fit options for selection These options include C Spline Linear Quadratic Cubic 4 P 2 P Logit Log PT to PT point to point and None Linear curve fit A simple best fit straight line 1s plotted using the values of the standards Quadratic or Quad curve fit A curve fit which 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 bx 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 standar
70. UNCTION list at the cursor position The available functions are LOG Log ELx808 Operator s Manual 3 35 EXAMPLES LOG10 Log Base 10 ALOG Anti Log Log10 2 0 301029995 ALOG 0 69314718 2 ALOG10 Anti Log Base 10 LOG Log ALOG10 0 30102995 2 LOG 2 0 69314718 AB Absolute Value SQRT Square Root AB 1 1 SORT 2 7 1 4142 PWR Power 10 PWR 2 100 e Well locations well types or numbers in parentheses precede functions e Press the CLEAR key to clear the item displayed at the cursor position e Press the SHIFT and CLEAR keys to clear the entire entry e 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 NUMBER OF REQUIRED CONTROLS BLANKS 3 36 If a control or blank validation formula is entered this screen allows you to enter the number of valid controls blanks for the assay Any previously defined values will appear on the display P C NUMBER OF VALID REPLICATES REQUIRED A e 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 e Press the CLEAR key to clear the displayed value e Press the ENTER key to save the displayed value and advance to the next screen or use the PREVIOUS SCREEN key to move backward through the menu structure Operation CUTOFF FORMULAS A cutoff formula calculates a cutoff value that i
71. acing the standards controls and samples in only the odd columns e P ACROSS 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 ELx808 Operator s Manual 3 21 CONSTANT BLANK VALUE ENTRY 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 the CLEAR key to clear the value on the display e Press the ENTER key to save the value and advance 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 ENTER NUMBER O F BLANKS e Use the NUMERIC keys to enter the number of blanks The range is 0 to 48 e Press the CLEAR key to clear the Number of Blanks value from the display e Press the ENTER key to save the value and move to the next screen 3 22 Operation SELECTING A BLANK LOCATION The Blank Location screen allows you to define where the blank well is located on the microplate This screen only appears 1f manual map generation ha
72. ad capabilities at the 100 light level and can help to identify deteriorating interference wavelength filters and other optical problems Perform these steps do not place a microplate on the carrier 1 For each filter in the filter wheel run a simple endpoint protocol and then print the results Do not apply blanking or transformations 2 Examine the OD values on the printouts Every OD should read 0 000 0 004 OD Note These limits are required by Bio Tek and cannot be changed 3 Sign and date the printouts and store them with your test documentation If the Empty Carrier test fails at more than one wavelength e Check the microplate carrier to ensure that it is clear of debris e Follow the steps on page 4 28 to clean the lamp contacts Verify that the lamp is properly aligned and then run the test again If the lamp has been in use for approximately 600 hours it may need to be replaced e Follow the steps on page 4 29 to remove the filter wheel and clean the filters If the Empty Carrier test fails at just one wavelength e Remove the filter wheel and examine the filter that exhibited the problem it may need to be replaced contact Bio Tek Check for spotting or a halo effect 4 16 Performance Verification and IQ OQ PQ Tests Liquid Testing Conducting Liquid Tests confirms the ELx808 s ability to perform to specification with liquid samples Liquid testing differs from testing with the Absorbance Test Pla
73. alification Procedures Y our ELx808 reader was fully tested at Bio Tek prior to shipment and should operate properly upon initial setup The tests outlined in this section may be utilized to confirm initial and ongoing performance of the ELx808 Set up the reader according to the Installation Qualification instructions on page 4 3 and confirm that it powers up and communicates with any peripherals After Installation Qualification conduct the Operational Qualification tests System Test Verifies proper gains bulb operation and low electronic noise Checksum Test Compares the basecode software to the internally recorded checksum values to ensure that the code has not been corrupted Displays software part number and version information Absorbance Plate Test Confirms the optical accuracy linearity and repeatability alignment channel to channel variation and wavelength accuracy of the instrument gt Accuracy of the optical density OD readings The comparison of the OD readings with those given with the Absorbance Test Plate insert will confirm the accuracy of the optical density values at that wavelength Alignment of the plate carrier and standard microplates 1s confirmed by the positional accuracy check of wells F01 C12 and H08 gt Channel to channel variation can be tested by completing the turnaround test This tests the reader s ability to read the glass filters accurately in different plate positions Lineari
74. alimentation Ein Verbindung mit dem Netz Conectado Chiuso Off Supply Arr t alimentation Aus Trennung vom Netz Desconectado Aperto sconnessione dalla rete di alimentazione Warning risk of electric shock Attention risque de choc electrique Gef hrliche elektrische Schlag Precauci n riesgo de sacudida el ctrica Attenzione rischio di scossa elettrica Warning risk of crushing or pinching Attention risque d crasement et pincement Warnen Gefahr des Zerquetschens und Klemmen Precauci n riesgo del machacamiento y sejeci n Attenzione rischio di schiacciare ed intrappolarsi Warning hot surface Attention surface chaude Vorsicht hei e Oberfl che Precauci n superficie caliente Attenzione superficie calda Caution refer to accompanying documents Attention voir documents d accompanement Achtung siehe Begleitpapiere Atenci n vease los documentos incluidos Attenzione consultare la doc annessa Separate collection for electrical and electronic equipment Les quipements lectriques et lectroniques font l objet d une collecte s lective Getrennte Sammlung von Elektro und Elektronikger ten Recogida selectiva de aparatos el ctricos y electr nicos Raccolta separata delle apparecchiature elettriche ed elettroniche Preface Intended Use Statement e The intended use of this instrument is dependent on the instrument s rear panel label If there is an IVD label then the instrument may be use
75. and S shall return an additional status string at the very end of the read sequence See Status String Format for error descriptions GET CURRENT STATUS 0 B 12 This command returns the most recent error status set by the instrument In the event of a reader timeout or DLE receipt it can be used to determine the error code associated with the reader failure It can also be used in place of pressing a key on the reader to stop the beeping after an error The status is always returned in the ELx status format host valid limits response o no arguments lt ACK gt status string 5 Requesting the current status through the use of this command will cause the status maintained on board the instrument to be cleared except in the following situations 1 A fatal error has been detected The only way to clear this type of error is by recycling power on the instrument 2 An error has been detected which requires a self test to be executed before a read operation can be performed In this case only a successful self test will cause the error to be cleared This command is most useful when invoked in pairs The first try will yield the most recent error detected or no error which will then be cleared by the reader if possible The next call will then yield either no error or a previously detected error that still needs to be cleared by running a self test Commands other than read commands may always be sent regardl
76. and Curve The DEFINE Option screen allows you to edit the Method Map Formula or Curve Fit FORMUTLA CURVE Press the soft key beneath the displayed option to access the following functions METHOD SOFT KEY 1 METHOD is selected and the user is prompted to select the read method parameters SOFT KEY 2 MAP is selected and the user is prompted for plate mapping information SOFT KEY 3 FORMULA is selected and the user is prompted to enter a formula SOFT KEY 4 CURVE FIT is selected and the user is prompted for curve fit options In addition the MAIN MENU PREVIOUS SCREEN and ENTER keys are active allowing you to move back and advance through the menu structure The definition of a method includes selecting Endpoint Kinetic or Scanning Read Modes Delay First Read Incubation Parameters Filter Wavelengths Applied Shake Parameters Kinetic Analysis ELx808 Operator s Manual 3 7 Note Some screens shown below and on the following pages may not appear on some reader models READ TYPE This option allows you to enter which read type Endpoint Kinetic or Scan The following keys are active during this screen READ TYPE KINETIC ENDPOINT KINETIC SCAN e Press SOFT KEY 1 to select Endpoint read mode Press SOFT KEY 2 to select Kinetic read mode e Press ENTER to save the displayed value and advance to the next screen DELAY IN FIRST READ Selecting the Delay in First Read option allows you to
77. ard Time EST or Customer Service between 8 00 AM and 5 30 PM EST Monday through Friday excluding holidays Technical Assistance Center 800 242 4685 Bio Tek Instruments Main Number 802 655 4040 Electronic Communication Send us e mail or visit our website E Mail tac biotek com Internet Site www biotek com ELx808 Operator s Manual 1 7 Fax Support 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 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 1 8 Ship instruments needing service to Bio Tek Instruments Inc Technical Assistance Center 100 Tigan Street Highland Park Winooski Vermont 05404 USA Note Please call or send e mail to TAC for an RMA number before shipping the instrument Introduction Chapter 2 Installation This chapter includes instructions for unpacking and setting up the ELx808 and instructions for connecting printers and or serial devices This chapter contains the following sections Unpack and Inspect the Instrument essesssssseseeeeeeenere nennen nnns 2 2 Select the Operating Environment 2 3 Install the Eilter Wheels bett eto he et e ertt nee 2 4 Check Adjust the Line Input Voltage Range ooooono
78. are accessed from the outer left side of the instrument Using a flat blade screwdriver loosen do not remove these two screws Slide the bracket out of the instrument Remove the bulb from its socket by pushing the socket and the bulb together and rotating the socket counterclockwise Rub the contacts with a pencil eraser two in the socket and two on the bulb end You can also use a cotton swab moistened with isopropyl alcohol Replace the bulb in the socket Replace the lamp assembly in the instrument Plug the lamp connector back into the circuit board Keep the shroud open to inspect and clean the wavelength filters see next page If cleaning the lamp contacts does not improve lamp output replace the lamp PN 3400508 See the instructions on page 4 32 The lamp on average should provide 500 hours of total service before it needs to be replaced This equates to approximately six months to one year of service under routine conditions Performance Verification and IQ OQ PQ Tests Inspecting and Cleaning the Wavelength Filters Laboratory air is used to cool the lamp and the wavelength filters can become dusty as a result The filters must be inspected and cleaned at least every three months To inspect and clean the wavelength filters 1 If you have not already done so turn off and unplug the reader and then follow the steps on the previous page to open the reader s top shroud 2 The filter wheel is located in the left
79. 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 with varying concentrations The assay data reduction states e Subtract the mean of CTL1 from the mean of the NC Subtract the difference from all ODs on the plate e Divide the result of the above by the means of the NC less the means of CTL1 and then multiply by 100 On paper the formula reads OD NC x CTL1 x NC x CTL1 x 100 e On the reader define the formula NC x CTL1 x as the Transformation Variable since the transformation will apply to all standards controls and samples on the plate e Atthe SCOPE VARIABLE screen choose OD and press ENTER Now enter the formula NC x CTL1 x by using the MATH OTHER MAP and FUNCTION keys Press the ENTER key e The formula definition screen is displayed Choose TRANS e Enter the following formula OD TVar TVar 100 using the MATH OTHER MAP and FUNCTION keys TVar is included in Map options on the formula entry screen The transformation formula has been added to the assay definition ANOTHER TRANSFORMATION 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 e Atthe SCOPE VARIABLE screen choose OD and press ENTER e Select STD1 from MAP and Press ENTER e Choose
80. ate light levels low electronic noise adequate photodiode sensitivity overall system cleanliness and 1f equipped proper function of the incubator The testing is designed to verify that the reader will give in specification performance for each set wavelength over the specified OD range If a printer is connected to the reader the test results will automatically print Note This test can also be run through KCjunior or KC4 running on a host PC Consult the appropriate user guide for instructions If the test passes the reader will beep once and the display will show the software s main menu which will resemble the following 1 30 PM DEFINE REPORT UTIL If the test fails the reader will beep repeatedly and the display will show an error code If this happens write down the error code and then press the Stop key on the keypad to stop the beeping Look up the error code in Appendix C Error Codes to determine its cause If the problem is something you can fix for example if the error code is 0201 or 0301 indicating that the filter wheel is missing turn off the reader fix the problem and then turn the reader back on If the cause is not something you can fix contact Bio Tek s Technical Assistance Center ELx808 Operator s Manual 2 13 Check Adjust the Reader s Filter Table After installing the filter wheel or new filters ensure that the ELx808 s filter table accurately maps the physical location of the filters
81. ate whether prompts shall be dispayed at run time for Plate IDs Sample IDs and Sample Counts e TESTS Access functions to test reader optics and instrument calibration Check the version of basecode software installed See Chapter 4 for more information To select the SETUP OUTPUT and READ user configurable options e Atthe Main Menu screen press the soft key beneath UTIL to access the Utility Options menu SELECIT UTILITY OPTION TESTS SETUP OUTPUT READ SETUP Options e Atthe UTILITY OPTIONS screen press the soft key beneath SETUP EDIT SETUP INFORMATION DATE TIME FILTER MOR E e Atthe EDIT SETUP INFORMATION screen press the soft key beneath DATE 10 09 04 DDMM Y Y e Enter the new date using the NUMERIC keys The cursor is positioned under the first editable field and advances automatically To change the date format press the soft key beneath MMDDYY or DDMMY Y The display updates to reflect the new format e Press ENTER to return to the EDIT SETUP INFORMATION screen ELx808 Operator s Manual 2 15 11PM 2 4HOUR e To edit the TIME press the soft key positioned beneath TIME At the TIME entry screen use the NUMERIC keys to enter the correct time e Selecta 12 or 24 hour format by pressing the soft key beneath these options The display automatically updates with the new time format e Press the Previous Screen key to return to the SELECT UTILITY OPTION menu OUTPUT Options e A
82. ated in the rear left corner of the instrument Unplug the lamp s connector from the main circuit board 3 Loosen the two flathead screws that retain the lamp bracket These are located on the left side of the instrument and are accessed from the outside of the instrument 4 Slide the lamp assembly forward and pull it in to the interior of the instrument then lift it out 5 The lamp is held in place by a spring To remove it pull on the back of the bulb and rotate the bulb toward the opening in the retention spring 6 Remove the bulb from its socket by pushing the socket and the bulb together and rotating counterclockwise then separating the bulb from the socket 7 Before installing the new bulb clean the lamp contacts with isopropyl alcohol and a cotton swab An alternative cleaning technique would be to rub the lamp contacts with a pencil eraser To install the lamp reverse steps 1 through 6 Keep your fingers out of the reflector interior and away from the bulb Ensure that the bulb is set flat against its housing before reinstallation in the instrument ELx808 Operator s Manual 4 33 4 34 Performance Verification and IQ OQ PQ Tests Appendix A Decontamination This appendix contains a procedure for decontaminating the ELx808TM It also contains a procedure for routine cleaning of the instrument s exposed surfaces This appendix contains the following sections PULPOSE 5 sete tede eel aan Bessel a ee pee sn Meade eee
83. athematical computations described below may then be performed and the template kept for future data reduction CALCULATIONS 7 Calculate the mean OD value for each well location in columns 1 and 2 for the five plates read in the Normal position and then again for the five plates read in the Turnaround position This will result in 32 mean values 8 Perform a mathematical comparison of the mean values for each microwell in its Normal and Turnaround positions that is compare Al to H12 Bl to G12 H1 to A12 and so on To pass this test the differences in the compared mean values must be within the accuracy specification for the instrument For example If the mean value for well A1 in the Normal position is 1 902 where the specified accuracy is 1 0 0 010 OD then the expected range for the mean of the same well in its Turnaround H12 position is 1 873 to 1 931 OD 1 902 0 010 0 010 0 029 1 902 0 029 1 873 1 902 0 029 1 931 4 20 Performance Verification and IQ OQ PQ Tests Note If any set of mean values is out of the expected range review the other three sets of mean values for the same channel pair For example if the A1 H12 comparison fails the wells are not within the expected range of each other review the comparisons of A2 H11 H1 A12 and H2 A11 If two or more sets of mean values for a channel pair are out of the expected range there is a problem with one of the eight read channels
84. ause the instrument to fail internal self checks e Excessive ambient light Bright sunlight or strong incandescent light can reduce the linear performance range and affect the instrument s readings e Dust Optical density readings may be affected by extraneous particles such as dust in the microplate wells A clean work area is necessary to ensure accurate readings ELx808 Operator s Manual 2 3 Install the Filter Wheel The filters that are shipped with the ELx808 are installed in the six position filter wheel For example the standard models have 405 450 490 630 nm filters the UV model s filter set is 405 450 490 630 and 340 nm The filter wheel which is packaged in a shipping envelope must be installed before the reader is used If you plan to install additional filters or change the filter locations use the following instructions to gain access to the filter wheel Plate access door 3 HJ Filter wheel shown installed Figure 2 2 Installing the Filter Wheel e If the reader is on turn it off and disconnect the power cord e Remove the seven screws around the perimeter of the shroud with a flat blade screwdriver as shown above Tip Bring the reader to the edge of the work surface to access the screws without having to turn the reader upside down e Carefully lift up the shroud from the front Note that the shroud is hinged along its back edge Hold the plate access door steady or tape it c
85. avelength Range 380 to 900 nm Filters 10 nm half bandwidth interference filters User accessible filter wheel Up to 6 filters may be installed on the instrument at one time Filters supplied 405 nm 450 nm 490 nm 630 nm and two blank filters Absorbance Measurement 0 000 to 4 000 OD Range e Single wavelength endpoint measurements with a 12 second read regular read mode Accuracy 1 0 0 010 OD from 0 000 to 2 500 OD 405 nm Linearity 1 0 from 0 000 to 2 500 OD at 405 nm 2 0 from 2 500 OD to 3 500 OD 405 nm Repeatability STD 0 5 0 005 OD from 0 000 to 2 500 OD 405 nm 1 5 0 005 OD from 2 500 to 3 500 OD 405 nm 2 5 0 005 OD from 3 500 to 4 000 OD 405 nm e Single wavelength kinetic measurements with read intervals of less than 12 seconds Accuracy 2 0 0 010 OD from 0 000 to 2 500 OD 405 nm Lineanty 2 0 from 0 000 to 2 500 OD 405 nm Repeatability STD 1 0 0 010 OD from 0 000 to 2 500 OD 405 nm Plate can be read at 6 second intervals rapid mode with wavelengths higher than 400 nm 1 4 Introduction e Display e Light Source e Dimensions e Weight e Environment e Humidity e Power Consumption Ultraviolet UV Model The following specifications apply only to 96 well flat or round bottom microplates 2 x 24 character LCD Tungsten halogen filled bulb 40 6 cm x 39 37 cm x 22 2 cm 16 0 deep x 15 5 wid
86. b Errors 200 and 201 indicate that a motor was not able to move to its home position as registered by feedback from an optical sensor PROBABLE CAUSE e Carrier movement may be limited so that the optical sensor cannot be interrupted Check the carrier path for obstructions e Filter wheel not installed The wheel is shipped in a separate envelope and may not be installed or may have broken free of the magnetic hub e Filter wheel movement may be impeded Remove the wheel to confirm if a loose filter is impeding the movement of the wheel Displayed Error Potential Cause ERROR 0300 Carrier failed to find light beam ERROR 0301 Filter wheel did not find home Errors 300 and 301 indicate that a particular axis has moved to a point where the light beam from the optics is no longer detectable by the measurement electronics This error is usually only seen during the service Auto Cal sequence PROBABLE CAUSE e Carrier Lamp may be out or blocked by a faulty filter wheel A loose belt or a loose motor pulley may cause the carrier to ignore movement instructions e Filter Wheel Motor The filter wheel is missing or loose or a loose filter may be impeding filter wheel movement Appendix C C 3 Displayed Error Potential Cause ERROR 0400 Carrier axis failed position verify ERROR 0401 Filter Wheel motor failed position verify Errors 400 and 401 indicate that an axis failed its Position Verify test After moving a prede
87. can be removed Ifthe 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 E 10 Instructions for Programming a New Assay 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 International Anticardiolipin Workshop Criteria Range GPLU ml High Positive gt 80 Medium Positive gt 20 80 Low Positive gt 10 lt 20 Results Interpretation The association between low positive levels of anticardiolipin antibodies and clinical findings is unclear Normal population studies indicate that there is a higher prevalence of IgM positives in the normal population than IgG 9 4 and 6
88. commendations included in the package insert for the test to be conducted Failure to conduct Quality Control checks could result in erroneous test data Introduction Hardware Features e Fight optics channels with an additional reference channel e A wavelength range of 380 900 nm standard model 340 900 nm UV model e Auser accessible 6 position filter wheel e A2 x 24 character LCD display e A membrane keypad with alphanumeric keys e Adjustable plate shake frequency and times e Reads 96 well microplates with 0 355 well centers e Operates from 100 120 230 or 240 VAC 50 to 60 Hz e One serial COM port 25 pin male connector and one parallel port 25 pin female connector e 4 Zone M incubation chamber option Software Features e Easy to use menu driven interface e Endpoint Kinetic and Linear Well scanning calculations e Curve fitting with 4 parameter cubic quadratic linear 2 P cubic spline and point to point methods e Transformation and formula calculations for more complex mathematical operations including validation and cutoff formulas e 55 assays are available onboard up to 75 custom assays can be preprogrammed and exported to the reader using Bio Tek s Define Protocol software e Automatically stores results for the last 10 plates ELx808 Operator s Manual 1 3 Specifications Standard Model Note The following specifications apply only to standard 96 well flat or round bottom microplates W
89. creen Enzyme Immunoassay Kit Transformation and Cutoff to translate the kit wording into an ELx808 assay program We are providing the following programming example using a fictitious n Vitro Diagnostic IVD assay These instructions are not intended to promote the use of the ELx808 with this or any other specific IVD assays and they do not constitute a claim of IVD use in jurisdictions where there are additional regulatory requirements For clarity only the user menu choices from the reader screens are shown Refer to Chapter 3 for details D Important The kit instructions are provided so that users can see how it is possible Intended Use This assay is designed for the in vitro screening of 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 autoantibodie
90. ctions and directions for programming an assay General Edited and formatted text Modified appearance of display screens Standardized the presentation of significant digits Changed Abs to OD throughout Updated the cover with a current photo of the instrument Updated contact information and warranty Changed Automated Microplate Reader to Absorbance Microplate Reader throughout Changed Universal Test Plate to Absorbance Test Plate throughout Removed references to internal barcode scanner and R models Removed references to General Formulas with respect to open assays Updated specifications to match product spec rev D Added text from rev J1 insert Fuse Installation Replacement Updated the installation instructions in Chapter 2 to better reflect actual practice Updated the IQ OQ PQ PM steps in Chapter 4 to better reflect actual practice Created Appendix F to provide instructions for adjusting the line input voltage range and replacing fuses for instruments with an older style power input module than the one described in chapter 2 Document Conventions This manual uses the following typographic conventions Example Description A This icon calls attention to important safety notes Warning A Warning indicates the potential for bodily harm and tells you how to avoid the problem Caution A Caution indicates potential damage to the instrument and tells you how to avoid the problem
91. d absorbance values in column Y e 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 e Click OK to perform the analysis the results of which will be output in a separate sheet e Note Ifthe Data Analysis command is not available on the Tools menu you may need to install the Analysis ToolPak in 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 of at least 0 99 is considered adequate REPEATABILITY TEST B 1 Calculate the mean and standard deviation for the five readings taken above at each concentration Only one data set needs to be analyzed The well that shows the most variation for any concentration is selected for data reduction For each mean below 2 500 OD calculate the allowed deviation using the repeatability specification for a 96 well plate of 0 5 0 005 OD from 0 000 to 2 500 2405 nm The standard deviation for each set of readings should be less than the allowed deviation ELx808 Operator s Manual 4 23 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 00
92. d for clinical research and development or other non clinical purposes If there is no such label then the instrument may only be used for research and development or for other non clinical purposes e The ELx808 is an 8 channel automated benchtop general purpose Enzyme Immunoassay Analyzer that performs analyses of a variety of samples The performance characteristics of the data reduction software have not been established with any laboratory diagnostic assay The user must evaluate this software in conjunction with the specific laboratory assay This reevaluation must include the establishment of new performance characteristics for the specific assay e This system is designed for use with a variety of enzyme immunoassays Assay protocol variations are addressed by the developer of the ELISA test kit in accordance with the test kit s procedure A versatile curve fitting and statistical software program is preloaded on every ELx808 If using Extensions Define Protocol software up to 75 assay protocols and blanking patterns can be defined stored in memory and instantly accessed Plate templates and formulas are automatically combined with the protocol assay setup Data results may be printed out or sent to a computer running a Bio Tek software package such as KCjunior or KC4 for Microsoft Windows The ELx808 s onboard software provides e An easy to use menu driven interface e Endpoint curvilinear regressional and stat
93. d 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 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 ELx808 Operator s Manual 3 43 3 44 4 Parameter Logistic or 4 P A curve fitted to the standard values which is characterized by a skewed sigmoidal S shaped plot that eventually becomes asymptotic to the upper and lower standard values The 4 parameters are Left asymptote Right asymptote Slope and Value at the Inflection point This fit is most recommended for immunoassay data and is more exact than Logit Log 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 the soft key below the menu option MORE to display additional options The top line of the display updates to reflect this selection Press ENTER to save the selection and advance to
94. d until another command is sent Computer Control 312 Status Format If the middle character in the status string is not ASCII 0 an error has been detected and no results data is transmitted ELx Status Format If any character in the initial status string is not ASCII 0 an error has been detected and no results data is transmitted Otherwise if an error is detected during the read process the final status string reflects that error See Status String Format for error descriptions SET ASSAY DEFINITION V This command downloads to the reader an assay definition table to be used in subsequent plate reads or wellset reads S or d commands until a new definition replaces it either via serial control or by the user from the front panel host valid limits response y lt ACK NAK gt data bytes 1 don t care 2 7 assay name ASCII characters 8 29 don t care 30 subtraction 0x00 none 0x40 dual wavelength 31 encoded byte bits 0 3 0xOF 0 bit 4 0x10 1 shake 0 no shake bit 5 0x20 1 shake before every read 0 before first read only bit 6 0x40 1 continuous shake 0 timed shake bit 7 0x80 1 use total kinetic time 0 use kinetic read count 32 1D scanning points 1 31 must be odd 33 49 don t care 50 52 measurement wavelength 000 340 900 53 55 reference wavelength 000 340 900 56 read type 0 endpoint 2 kinetic 3 1D scan 57 59 don t
95. e 2 6 Note The information in this appendix formerly resided in Chapter 2 in ELx808 Operators Manual revisions J1 and earlier Adjusting the Line Input Voltage Range F 2 The ELx808 is equipped with a four voltage range power input module This power input module located on the right side of the instrument can be adjusted for 100 VAC 120 VAC 230 VAC and 240 VAC voltage inputs The setting can be determined visually by observing which indicator hole on the power input module has a peg within it The ON OFF switch and fuses are also housed within the power input module The following instructions can be used to change the input voltage range The figure below illustrates the power input module Instructions for Adjusting the Line Input Voltage Range e Unplug the reader and remove the power cord e Use a small flat blade screwdriver to pop the fuse holder out of the power input module Note Ifthe cover swings back from the instrument instead of popping out and the fuse holder is not attached to the cover your instrument has a newer style power input module Turn to page 2 6 now e A small voltage select switchboard is located on the right of the power input module The switchboard which can be removed with needle nose pliers can be oriented four ways to select four different voltage input ranges e Once the switchboard is removed text can be read on one side of the board This text identifies the switch positions The tex
96. e Accuracy 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 proven that the device is accurate at these OD values the device is also considered to be linear Note that there may not be a Pass Fail indication for filter values that are beyond the specified accuracy range of the instrument e Repeatability The Repeatability 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 instrument e Channel to Channel Variation This test ensures that selected channels read the same value for a filter as their paired channel when the plate is turned around 180 in the plate carrier It is a way to verify that each channel s Delta values are adjusted relative to the signal strength of the reference channel 4 10 Performance Verification and IQ OQ PQ Tests Requirements To run the Absorbance Plate Test you need Bio Tek s Absorbance Test Plate PN 7260522 or 9000547 with its accompanying Data Sheet shown below This test plate can be used for testing the reproducibility linearity and alignment of your Bio T
97. e Specifications sssssssseeeeeeeeee 4 11 Running the Absorbance Plate Test 4 12 Repeated Test Failures ccccccccccccssecsteceseceseceneceeeseeeeeseeeeeeeseeeseecsaecnseenseenseees 4 15 Empty Carrier Test uncos d aea RERO TR TRUE E ee 4 16 Liquid Testing secte es EISE CE NEG ERSTER ug Ted RE a Ses 4 17 Stock Solution Formulation esee nnne 4 18 Liquid Test 1 53 ph ads 4 20 Liquid Test 25 ds a tte m i editis 4 22 Liquid Lei er e ete n ERE re reb E ORE tee 4 25 Periodic Maintenance for the ELx808 oooooniccnicconoconococonocononononanonnn nono noon ccoo nono nonononns 4 28 Cleaning the Lamp Contacts eere 4 28 Inspecting and Cleaning the Wavelength Filters sess 4 29 Periodic Maintenance for ELx808 Robotic Units essere 4 30 Replacing the Lamp 52 rte It Ree hee 4 32 Recommendations for Achieving Optimum Performance 4 2 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 ELx808TM supports standard flat U bottom and V bottom 96 well microplates the reader achieves optimum performance with optically clear flat bottomed wells Non uniformity in the optical density of the well bottoms can cau
98. e a defective fuse pop out the old fuse and replace it with the correct new one The configuration of the fuse module U S or European is determined by which fusing network is facing the inside of the power input module To change the configuration remove the Phillips head screw that anchors the fuse holder Remove the fuse holder and turn it over Replace the screw and reinstall the fuse module in the power input module Appendix F F 3 F 4 Instructions for Adjusting the Line Input Voltage Range
99. e currently set SELECIT BAUD RATE 1200 2400 9600 e To change the BAUD rate press the soft key beneath the desired baud rate The display top line automatically updates to reflect the new choice e To view the reader s settings for parity stop bits and data bits press the soft key beneath VIEW 2 20 Installation Serial Port Pinout Description The serial port on the ELx808 is a DTE configuration with a 25 pin pin male D sub connector If the supplied serial port becomes lost or damaged contact Bio Tek for information on replacement cables Table 2 2 Serial Pinout Description Serial Pin Description Signal NC HS ELx808 Operator s Manual 2 21 After Installation and Setup Verify Performance Before using the ELx808 for the first time verify that the reader is operating properly by running a System Test and the Absorbance Plate Test These tests and additional verification procedures are described in Chapter 4 Before Repackaging the Instrument e Decontaminate the reader prior before shipping Refer to the Decontamination procedure in Appendix A e Once the reader is clean pack it in its original shipping box using original packing materials see Figure 2 1 on page 2 3 e Contact Bio Tek s Technical Assistance Center for a Return Materials Authorization RMA number and shipping instructions e This shipping system was designed to be used no more than five times If the container
100. e number of any predefined Assay Definition Files stored in the reader s memory or the OPTION key to advance one assay at a time The cursor is positioned at the first editable field and advances automatically The numeric range depends on the number of assays 1 55 programmed in the reader s memory The assay s name and number are displayed on the screen SELECIT ASSAY NUMBER NAME HBS AG e Press ENTER to advance to the Run Time prompts e MAIN MENU Returns the display to the Main Menu screen RUN TIME PROMPTS After the assay is selected you may be prompted for information depending on preferences selected in the reader s Utilities whether a manual map was set up or if the assay was created and downloaded from Bio Tek s Extensions amp Define Reader Protocol software 3 50 Operation If the assay was created using Extensions software prompts might include e The number of samples e Standard Concentrations e Assay ID e Fill Pattern e Blank Method e First Well Location e Replicate Count for each well type e Wavelength Mode e Report preferences etc Refer to the Extensions Define Protocol Software User s Manual for more information on user prompts that might be encountered Other READ prompts might include e Enter number of samples e Plate ID e Enter Sample ID If a Manual Map is used prompts for information might include e Well locations for each sample ELx808 Operator s Manual 3 51 ENTER NUMB
101. e power cord 2 Insert the needle nose pliers into the two holes on the cover of the power entry module Lift the cover open and swing it back into the socket Note If the cover pops out of the instrument instead of swinging back and the fuse holder is attached to the cover your instrument has an older style power input module Turn to Appendix F now 3 As shown above a small voltage selector card is located to the right of the fuse holder Use the pliers to pull this card straight out of the housing Installation 4 As shown below there are four different voltage input range positions Rotate the white plastic indicator around the card so that it fits into the correct groove for the desired voltage Note Point the indicator pin upward so that the text matching the desired voltage range is readable at the bottom of the card as shown below White plastic indicator pin N should point L upward This side showing text should face the ON OFF switch voltage range should be on the bottom Figure 2 4 Voltage selector card input range positions 5 Reinsert the voltage selector card into the power entry module so that the text side faces the ON OFF switch and the desired voltage range is on the bottom of the card see above 6 Reconfigure fuses as needed see next page 7 Replace the cover and verify that the white plastic indicator pin lines up with the correct power entry module indicator hole
102. e returned to their home positions the plate is moved back outside where it can be accessed 2 The reader transmits a lt DLE gt character to the computer when the above process is completed 3 No more data is transmitted by the read in progress A final status string is transmitted in the ELx Status Mode Appendix B B 23 Status String Format Following execution of each command except as noted the ELx808 sends a status string back to the computer 312 Status Mode Format This status string consists of 5 successive ASCII characters RS S1 S2 S3 and ETX gt RS A record separator that marks the beginning of the status string gt SI Always ASCII zero 0 gt S2 A single digit used as a reader fault or error code number ERROR CODES Error codes indicate the following 0 no fault or error 8 instrument failure perform self test 9 error in assay scan or table definition A error in well range selection B incubator setpoint error C incubator temperature error E barcode error gt S3 Always ASCII zero 0 gt ETX End Of Text marks the end of the status block Instrument failures are usually accompanied by an error code on the reader display ELx Status Mode Format This status string consists of 5 successive ASCII characters a four byte string representing a hexadecimal status code and then ETX Items described in angle brackets lt gt are ind
103. e x 8 75 high 15 87 kg 35 Ib maximum Operating temperature 18 to 40 C 10 to 85 noncondensing 100 VA four voltage ranges accommodated by the voltage selection switch Range 1 Range 2 Range 3 Range 4 100 VAC 120 VAC 230 VAC 240 VAC Wavelength Range 340 to 900 nm Filters Supplied 340 405 450 490 630 nm Optical specifications for the 400 to 900 nm range 12 second read in regular read mode Same as standard model Optical specifications for the 340 to 400 nm range 12 second read in regular read mode Absorbance Measurement 0 000 to 3 000 OD Range Accuracy 1 0 Linearity E Repeatability STD 1 0 Incubator Model The following specifications apply to a 96 well sealed plate with 200 ul of liquid in all wells e Temperature Control e Temperature Variation ELx808 Operator s Manual 90 to 110 VAC 50 to 60 Hz 108 to 132 VAC 50 to 60 Hz 207 to 253 VAC 50 to 60 Hz 216 to 264 VAC 50 to 60 Hz 0 010 OD from 0 000 to 2 000 OD 340 nm t 1 0 from 0 000 to 2 000 OD 340 nm 0 005 OD from 0 000 to 2 000 OD 340 nm Temperature controlled to 50 C 0 50 C 37 C with the plate sealed 1 5 Package Contents The contents of the ELx808TM package include Absorbance Microplate Reader Power Cord Filter wheel with four standard filters 405 nm 450 nm 490 nm 630 nm and two blank filters UV models also include a 340 nm filter Ope
104. ed 000 999 assay name 8 312 format only ABCDEF lt CR gt lt LF gt unused extra characters 49 312 format only wavelength terminator 1 lt Z gt checksum 1 ELx format only 0 255 final terminator 1 ELx format only lt Z gt final status string 5 ELx status mode only Appendix B B 13 Temperature response included only if requested Format is three ASCII bytes scaled up by ten 1 e 275 correponds to 27 5 degrees C See the Include Temperature Response command Absorbance data is returned in the form ofa 5 character ASCII string with a sign character followed by four digits A decimal point is not included but should be inserted by the user after the first digit 1 e 1234 should be translated as 1 234 OD Overrange readings are indicated by the 5 character string instead of a signed numerical value If dual wavelength subtraction has been selected in the reader definition table the subtraction is performed by the reader and the result is sent as a single plate representing the difference in ODs per well Otherwise a complete plate of ODs is sent for each wavelength selected If an error is detected during a plate read or the user aborts the process via serial X or stop key press the entire remaining data response is replaced by a single lt DLE gt character 0x10 In the ELx Status Mode the final status string then indicates the error detected In the ca
105. ed by the operator ELx808 Operator s Manual vii Precautions viii The following precautions are provided to help you avoid damaging the system Caution Service The system should be serviced by authorized service personnel Only qualified technical personnel should perform troubleshooting and service procedures on internal components Caution Environmental Conditions Do not expose the system to temperature extremes Ambient temperatures should remain between 18 to 40 C System performance may be adversely affected 1f temperatures fluctuate above or below this range Caution Sodium Hypochlorite Do not expose any part of the instrument to Sodium Hypochlorite solution bleach for more than 30 minutes Prolonged contract 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 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
106. eee A 2 T ols and Su pplies A ete ten e iet ies A 3 Decontamination Procedure esssssssssseseeeee eene A 3 Routine Cleaning Procedure ooooooninonooonnonononononnconnonn nono ncon nono nono naco E A S nennen A 5 lunc E A 5 Tools and Supplies ntm tete aet ledio tic e Re ss elas Ses A 5 Purpose 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 in contact with the instrument during shipping handling and servicing Decontamination is required by the U S Department of Transportation regulations Persons performing the decontamination process must be familiar with the basic setup and operation of the instrument D 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 and nose Eating or drinking while decontaminating instruments is not ad
107. ek or if the product has had the serial number altered effaced or removed This warranty is void if the instrument has not been connected installed or adjusted strictly in accordance with written directions furnished by Bio Tek Batteries fuses lamps and other consumable items used in any of the instruments are not covered by this warranty Bio Tek will continue to stock parts for a minimum of five 5 years after the manufacture of any equipment has been discontinued Parts shall include all materials charts instructions diagrams and accessories that were furnished with the standard instrument xiv Preface Table of Contents req m iii All Rights Reserved cios iii Trade mark tii x iii Restrictions and Liabilities sinni toda iii Revision HIStOFy irit oce to ED e dei Ne tedcedip isa iv Document Conventions cienia a id vi Warnings and Precautions ccoo e iea i auian vii Electromagnetic Interference and Susceptibility oooooooocononininicncnnnnonacoccccccncnananannncnncnannnn nos ix oalety te HR REPERI ix CE Mark Information 222 tierno Iure esu ate pee tt ea X Safety SYMONS iii sede xi Intended Use Statement nee gi ria cinta xiii Specimen Prepaid xiii Registration Card nte da eee eine xiii ECCE xiv Chapter 1 Introduction a ists vsccsceeicssecevvedecoescovsdoresservensvensvssessersveieseedecesderestareeit 1 1 Introduc
108. ek autoreader The following calibration data has been recorded by a N I S T traceable spectrophotometer WAVELENGTH nm Well 405nm 450nm 490nm 550nm 620nm 630nm 690nm 750nm C1 0 147 0 140 0 135 0 130 0 136 0 136 0 127 0 134 E2 0 618 0 575 9 568 0 485 0 434 G3 1 133 1 052 S A M P L E 040 0 881 0 783 H6 1 701 1 578 560 1 323 1 179 F5 2 279 2 024 1 976 1 956 1 893 1 865 1 537 1 272 D4 2 945 2 604 2 545 2 513 2 437 2 400 1 972 1 632 Set 2453 Serial 161259 Figure 4 2 Sample Absorbance Test Plate data sheet Entering the Test Plate Specifications Using the Data Sheet provided with the Absorbance Test Plate follow the steps below to enter the calibration values into the reader 01 30PM 10 09 03 DEFINE REPORT UTIL e From the Main Menu select UTIL gt SETUP gt MORE gt CAL PLATE e The CALIBRATION FILTER selection screen will appear CALIBRATION FILTER 405 450 490 e Select the filter you wish to enter data for ELx808 Operator s Manual 4 11 e A data entry screen will appear requiring the entry of a Well Location and corresponding Calibration Value Carefully enter the information from the Data Sheet WAVELENGTH 40 5 WELL CALIBRATION VALUES e After each entry press ENTER to advance to the next well location Continue to enter the values listed on the Data Sheet for each well location at each filter wavelength value you wish to test e When all values have been entered press the MA
109. em self test and calibration This should be performed any time the wavelength configuration is changed M command host valid limits response ve no arguments lt ACK NAK gt data stream status string 5 The reader responds by sending a variable sized stream of ASCII character data representing various calibration and test results This data stream is followed by the standard status response string SET TEMPERATURE This command sets the incubation chamber temperature setpoint host valid limits response T lt ACK NAK gt data bytes 1 2 temperature setpoint 0 22 50 status string 5 If the instrument is not equipped with a working incubator or if the indicated temperature setpoint is out of range an incubator setpoint error will be returned with the standard status response string A setpoint of zero will turn the incubator heaters off Appendix B B 19 GET TEMPERATURE J This command returns the current temperature in the incubation chamber host valid limits response Y no arguments lt ACK NAK gt temperature 2 status string 5 The reader returns the current temperature as a 2 byte integer value not ASCII so low byte first Temperature is returned scaled up by 10 1 e 370 indicates a temperature of 37 0 degrees Celsius Temperatures below the instrument minimum of 18 degrees are returned as 111 0x006F temperatures above the instrument maximum of 55 degrees are retur
110. ength kinetic measure ments will be read in intervals less than 12 seconds Regular or Normal Mode Specifications for these modes are outlined in Chapter 1 When selections are completed the display returns to the SELECT UTILITY OPTION screen Installation Set Up the Serial Port for Communication with Other Devices The ELx808 has a 25 pin serial RS232 port located on the rear panel of the instrument see Figure 2 5 on page 2 11 The serial port allows the reader to communicate with a computer using Bio Tek s KC4 or KCjunior software standard communications software and or RS 232 protocols Appendix B contains information on required protocols for computer control of the reader In addition Assay Definition Files created using Bio Tek s Microsoft Windows based Extensions Define Reader Protocol software can be downloaded to the reader using the instrument s RS 232 serial interface For more information about downloading custom assay files contact Bio Tek Attaching the Serial Cable If the ELx808 will be controlled by software running on a host PC attach the serial cable e Power down the computer and the reader e Connect the appropriate serial cable to both machines e Power up the reader and the computer e Ensure that the ELx808 and the computer are operating with the same communications settings see below Setting Communications Parameters Before serial communications are initiated between t
111. enter a time delay before the first read DELAY FIRST RE AD TIME e Enter the time in minutes and seconds using the numeric keys e Press ENTER to advance to the INCUBATION TEMPERATURE screen 3 8 Operation INCUBATION TEMPERATURE The incubation temperature screen allows you to set the assay incubation temperature INCUBATION TEMP 37 C AMBIENT TEMPERATURE e Press SOFT KEY 1 or 2 to select ambient incubation e Press SOFT KEY 3 or 4 to select an adjustable temperature e Use the LEFT and RIGHT ARROW keys to move the cursor between the two digits on the input temperature e Use NUMERIC keys to enter the incubation temperature up to 50 C e Press ENTER to save and advance to the next screen SINGLE OR DUAL WAVELENGTH The WAVELENGTH selection screen allows you to select SINGLE or DUAL wavelength for the assay WAVELENGTH SINGLE DUAL e Press SOFT KEY 1 to select SINGLE wavelength The reader will measure the optical density of each well with a single filter e Press SOFT KEY 2 to select DUAL wavelength Each well will be read twice each time with a different filter Note The microplate 1s not removed from the reading chamber between the two measurements The final reported optical density is the difference between the two readings Dual wavelength readings can significantly reduce optical interference caused by scratched or fingerprinted microplates since the scratches or fingerprints reduce the amoun
112. er Adjust it if necessary according to the directions in Chapter 2 Locate the power inlet on the side of the ELx808 Plug the rounded end of the power cord into the power inlet Plug the 3 prong end of the power cord into an appropriate mains power receptacle If the reader will be run in standalone mode that is without any controlling software running on a host computer connect the printer to the reader using the supplied parallel cable Turn on the printer Turn on the reader using the switch located on the right side Verify that the ELx808 s internal filter table matches the physical locations of the filters in the filter wheel UTIL gt SETUP gt FILTER Set the current time and date If a printer is connected set the data output and reporting options Select the proper printer type Set the read time prompts and read speed Run a System Test and print the results Run a Checksum Test and record the checksum part number s and version number s of the software See page 4 9 for instructions If the instrument will be controlled via software running on a host PC Connect the PC to the reader using the serial cable b Ifyou will be using KC4 or KCjunior install the software on the PC c Ensure that the communication parameters match on both devices e g the baud rate may need to be adjusted on the ELx808 d Launch the PC program and ensure communications are established Run a System Test and p
113. es 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 normal 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 E 4 Instructions for Programming a New Assay Programming the ANA Screen Enzyme Immunoassay Kit Transformation and Cutoff according to their specific kit instructions and for verifying that the calculations are performed correctly If you are not familiar with programming formulas onboard the ELx808 refer to Formula Entry in Chapter 3 D Important The ELx808 user is responsible for programming the reader properly From the Main Menu press DEFINE Enter the assay number and edit the name if desired At the DEFINE menu STEP COMMENTS 1 To program the reading method press METHOD READ TYPE Endpoint Kinetic or Scan Endpoint DELAY FIRST READ IF INCUBATOR MODEL INCUBATION TEMP Ambient or Temperature WAVELENGTH Single or Dual Single MEASURE Wavelength s to use 450 SHAKE First Every or None 2 To program the plate map press MAP AUTO Auto
114. ess of the current reader status if there is a hardware issue preventing successful execution a subsequent error status will be generated See Status String Format for error descriptions Computer Control SET QUIET MODE q This command allows the user to select a quiet mode When this mode is turned on the reader bypasses the process of displaying an error code and beeping until a key is pressed As long as the quiet mode is selected errors detected while processing serial control commands are available only if ELx Status Mode is selected or in response to the Get Current Status 0 command Serial only instruments are set to quiet mode automatically at powerup host valid limits response q no arguments lt ACK gt data bytes mode 0 off 1 on quiet status string 5 READ PLATE 8 This command causes a microplate to be read according to the currently loaded assay definition table host valid limits response Ss no arguments lt ACK NAK gt reader response protocol status string 5 for each kinetic interval one if endpoint read for each wavelength expected 312 format one ELx format up to three wavelength start code 1 lt CR gt for each row within specified row range for each column well within specified column range E comma separator 1 sign 1 or data 4 1234 row terminator 2 lt CR gt lt LF gt temperature 3 if specifi
115. etic assay with a previously specified shake has been selected SHARKE TIME CONTINUOUS e Use the NUMERIC keys to enter the shake interval Valid ranges are 0 1 hours 0 59 minutes and 0 59 seconds e Use the LEFT and RIGHT ARROW keys to move the cursor between hours minutes and seconds e Press ENTER to save the entry and advance to the next screen SHAKE SPEED Use this screen to select the shake speed The shake movement is a repeated 0 021 inch movement from the shake position and back SHAKE SPEED MEDIUM LOW MEDIUM H I GH VARI e Press SOFT KEY 1 to select low speed shaking e Press SOFT KEY 2 to select medium speed shaking e Press SOFT KEY 3 to select high speed shaking e Press SOFT KEY 4 to select variable speed shaking 1 second of each speed repeated e Press ENTER to save the entry and advance to the next screen ELx808 Operator s Manual 3 13 KINETIC DATA ANALYSIS SELECTION Use this screen to select the type of kinetic analysis KINETIC ANALYSIS RATE R SQR ONSET e Press SOFT KEY 1 to select the kinetic rate calculation This method will apply a linear fit to calculate the maximum slope based on the number of kinetic points specified e Press SOFT KEY 2 to select the R squared rate calculation This method will calculate the R squared value at the maximum slope based on the linear curve fit and the number of kinetic points specified e Press SOFT KEY 3 to select the time calculation
116. fined number of steps from a home position the motor should return to the home position in the proper amount of time and steps If the axis moves back to its home position in the wrong amount of time or in too few or too many steps the test falls PROBABLE CAUSE e Mechanical obstruction e Carrier belt or pulley slipping e Loose filter wheel hub e Loose filter may be impeding filter wheel movement e Defective motor drive circuit or 12 volt power problem Displayed Error Potential Cause ERROR 0500 0502 0506 Filter is not installed Error 500 indicates an empty filter location on the filter wheel or that the filter wheel is not installed PROBABLE CAUSE e All filter locations must have either a filter or a filter blank BTI PN 3122037 installed The reader may also display this error if the filter wheel is not installed C 4 Error Codes Displayed Error Potential Cause ERROR 0601 to 0606 FILTER 1 to 6 Gain out of range Errors 601 606 indicate that the gain for a specific filter is out of range Gain out of range indicates that the lamp is out or that a particular filter absorbs or transmits entirely incorrect light levels PROBABLE CAUSE e Lamp out Check that the lamp is lit when the unit is ON Replace lamp if necessary e A missing defective or misplaced filter or filter blank will cause this error Observe that the filters in the wheel match the filters programmed in the software Look for obviou
117. formation variable Tvar FORMULA CTL1 X Defining TRANS VAR CTL1 X isolates the OD value for CTL1 X for use in transformation TRANS FORMULA OD TVAR 10 Converts all OD values on plate to ANA Result per kit insert instructions 4 To define a cutoff formula for Positive and Negative calls FORMULA CUTOFF 10 0 GREYZONE 00 SAMPLE gt CUTOFF POS Appendix E E 7 Sample Anticardiolipin IgG Enzyme Immunoassay Kit Standard Curve and Cutoff Important The kit instructions are provided so that users can see how it is possible to translate the kit wording into an ELx808 assay program We are providing the following programming example using a fictitious n Vitro Diagnostic IVD assay These instructions are not intended to promote the use of the ELx808 with this or any other specific IVD assays and they do not constitute a claim of IVD use in jurisdictions where there are additional regulatory requirements For clarity only the user menu choices from the reader screens are shown Refer to Chapter 3 for details Intended Use This assay is intended for the in vitro measurement of IgG anticardiolipin antibodies in serum as an aid in the diagnosis of antiphospholipid syndrome APS Sufficient materials are supplied to allow a maximum of 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
118. he ELx808 and another device the communication parameters Baud Rate Data Bit configuration and Parity status must match on both devices The reader s default communication parameters are e 2400 Baud e 8 Data Bits e 2 Stop Bit e No Parity The user may change the Baud rate on the reader from the recommended default setting of 2400 to 1200 or 9600 see instructions on the next page Data bits Stop Bits and Parity are not user configurable The communications software operating on the host computer device should be set to 8 2 and None Refer to the KCjunior or KC4 user guide for additional information on setting testing communication parameters with the ELx808 ELx808 Operator s Manual 2 19 Changing the Baud Rate on the ELx808 To change the baud rate from the default of 2400 to either 1200 or 9600 e Turn on the instrument if it is not already on 01 30PM 10 09 04 DEFINE REPORT UTIL e Atthe Main Menu screen press the soft key beneath UTIL SELECIT UTILITY OPTION TESTS SET UP OUTPUT READ e Atthe UTILITY OPTIONS screen press the soft key beneath SETUP EDIT SETUP INFORMATION DATE TIME FILTER MOR E e Atthe EDIT SETUP INFORMATION screen press the soft key beneath MORE to advance to the EDIT SETUP RS232 option screen EDIT SETUP INFORMATION RS2 32 CALPLATE MOR E e Press the soft key beneath RS232 to access the BAUD RATE selection menu The top line of the display shows the baud rat
119. he Number of Assays entry screen will appear ELx808 Operator s Manual 3 47 e Specify the number of assays to include in the panel 1 to 8 e Press ENTER to continue The Mapping Direction selection screen will appear MAPPING DIRECTION DOWN DOWN ACROSS e This option ensures that all assays will be mapped in the same direction e Select DOWN or ACROSS Note The original mapping directions for the predefined assays are overridden by the Panel s mapping direction If the assays include 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 HBS AGI1 e Press OPTION to cycle through the assay numbers and names or use the NUMERIC keys to enter an assay number Press ENTER to make a selection e After selecting an assay you must define its starting location 3 48 Operation 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 through A12 are valid for DOWN mapping e This process will be repeated for each assay within the panel Remain aware of the total number of controls standards and blanks that were originally mapped in each individual assay while mapping for the Panel assay e For example to include Assays 1 8 and 22 in the Panel assay DOWN
120. he power inlet on the instrument 3 Plug the 3 prong end of the power cord into an appropriate mains power receptacle A Warning Power Rating The ELx808 line power 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 Electrical Grounding Never use a two prong plug adapter to connect primary power to the ELx808 Use of a two prong adapter disconnects the utility ground creating a severe shock hazard Always connect the line power cord directly to a three prong receptacle with a functional ground Installation Install the Printer If the ELx808 will be operated in standalone mode that is without Bio Tek s KCjunior or KC4 software running on a host PC connect a printer directly to the reader using the supplied parallel cable To attach a printer to the ELx808 Important To avoid system instability make sure the printer and reader are turned OFF before connecting them e Ifthe reader and or printer are on turn them off Place the printer in a location adjacent to the reader e Attach one end of the parallel cable to the printer s parallel port e Attach the other end of the cable to the reader s parallel port located on the instrument s rear panel e Make sure the securing screws on both ends of the cable are tightened
121. icated by an ASCH digit replacing the last 0 character in the status code Items described in curly braces are indicated by an ASCII digit replacing the next to last 0 character in the status code Computer Control Fatal errors indicate a hardware failure also shown on the instrument display screen and require recycling of instrument power Note Errors listed below are common to all reader instruments and may not all be applicable to any single given reader Fatal Errors TCB NOT AVAIL ERR READ NOT AVAIL ERR NOT AVAIL ERR CHECKSUM ERR DR ALLOC ERR DFLASH TIMEOUT ERR DFLASH ERR CFLASH TIMEOUT ERR HEAP CORRUPTION ERR Non Fatal Errors NO ERR ABORT ERR NO SENSOR ERR NO BEAM ERR MOTOR VERIFY ERR SATURATION ERR FILTER GAIN ERR NOISE TEST ERR OFFSET TEST ERR DARK RANGE ERR 0900 AIR RANGE ERR ASSAY NUM ERR PRINT TIMEOUT ERR CAL CHECKSUM ERR WAVE NOT FOUND ERR FILTER SIGNAL ERR CNFG DATA ERR CNFG CHECKSUM ERR CAL DATA ERR MOTOR NOT HOMED ERR INCUBATOR FAILURE SC ASSAY DEF ERR KIN INTERVAL ERR KIN COUNT ERR MALLOC ERR Appendix B A100 A200 A300 A400 A500 A600 A700 A800 A900 0000 0100 0200 0300 0400 0500 0600 0700 0800 task control block not available read already in progress lt device gt not available failed code checksum test on powerup DR
122. ight is influencing the readings Because of the mathematical correction this difference under normal conditions should be slight or nonexistent A 10 minute warm up of the instrument is suggested prior to reading to achieve the best repeatability from microplate to microplate measurements Performance Verification and IQ OQ PQ Tests Installation Qualification IQ The IQ is conducted when unpacking and setting up the ELx808 according to the following steps Refer to Chapter 2 Installation where necessary for illustrations and additional information l 10 11 12 13 14 15 Unpack the instrument as outlined in Chapter 2 and ensure that all required items and any purchased optional items are present Check for any possible shipping damage Record the Instrument Model Number Serial Number Bio Tek Sales Order Number and the date of shipment from Bio Tek Locate the reader on a level stable surface where ambient temperatures between 18 C 65 F and 40 C 105 F can be maintained Environmental conditions to avoid include excessive humidity excessive ambient light and dust Install the reader s 6 position filter wheel as described in Chapter 2 This involves removing the perimeter screws from the reader s top shroud to access the internal components Record the filter wavelength values and their locations in the wheel Check the voltage setting on the power entry module located on the right side of the read
123. ill be displayed This option screen appears only if Manual Map Generation has been selected AXIS TYPE LIN LIN LOG LOG LOG LIN e Press SOFT KEY 1 2 3 or 4 to select the method by which the X and Y axes will be scaled The top line of the display updates to reflect the selection 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 e Press ENTER to save the selection and advance to the next screen 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 Any previously defined decision appears on the screen EXTRAPOLATIE UNKNOWNS YES NO e Press SOFT KEY 1 to select YES extrapolate the unknowns The top line of the display updates to reflect this selection e On the printed reports extrapolated concentrations RSLT values are surrounded by lt gt e g lt 44 425 gt e Press SOFT KEY 2 to select NO The top line of the display updates to reflect this selection 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 D
124. ing the EEXx8908 e epe ide ded ite e eee ee 1 2 Quality Control 52 622 tt dde ee Ee 1 2 Hardware Features 2 oto p ede lide dete fox teri eii ix e DUI dne esp de trips 1 3 Softwate FeatUr s idc e e qc n ima ei ee ei tete dp IEEE De aa 1 3 Specifications mee EE 1 4 Standard Model D 1 4 Ultraviolet UV Model 1 treten trennen needle ede nete ede tene E cine ct 1 5 Incubator Model rrr et arena RP 1 5 Package Contents aa 1 6 elei Baee cconccpe RT 1 6 Technical SUP POM E 1 7 Chapter 2 Installation eee ecc 2 1 Unpack and Inspect the InstruMeNt ocooonoccccnnocanonoccncnanoncnn nono nnn nao n nn nano nn nr nana nn nan nnr narran 2 2 Select the Operating Environment essen erem 2 3 Install the Filter WLieel 2 iiic ERR as 2 4 ELx808 Operator s Manual xvi Check Agjust the Line Input Voltage Range ssesee mee nennen 2 6 Reconfiguring or Replacing Fuses ssses eene enm enn 2 8 iere dae dc Em 2 10 Install the Printer terere en tee ree eerte e Eee does 2 11 Printers ir pe e ea e Aa ee ined setts 2 12 Cables cunt nn e eua uU M E Montt dde acre Ai MALE eA el ds 2 12 Parallel Port Pin Definition sni e rer HC Pie REP ER Een 2 12 Turn on the Reader and Run a System Test ssssssssssseeee emen 2 13 Check Adjust the Reader s Filter Table sssee enm 2 14 Config
125. ing the ingredients below Table 4 5 Phosphate Buffered Saline 10X Concentrate Solution NaCl KCI neo oo KO rams 2 Mix 5 ml of the stock buffer solution with 45 ml of deionized water 3 Add 10 mg of the B NADH powder and mix thoroughly This is the 10x Concentrate PBS Solution ELx808 Operator s Manual 4 25 4 26 SOLUTION B SIGMA PBS Deionized water Phosphate Buffered Saline PBS pH 7 2 7 6 Sigma tablets P4417 or equivalent B NADH Powder f Nicotinamide Adenine Dinucleotide Reduced Form Sigma bulk catalog number N 8129 or pre weighed 10 mg vials Sigma number 340 110 Note Store the B NADH powder according to the guidelines on its packaging Prepare a PBS solution from the Sigma tablets In a beaker mix 50 ml of the PBS solution with 10 mg of the B NADH powder and mix thoroughly This is the Sigma PBS Solution PROCEDURE 1 Check the absorbance of a sample of either buffer solution 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 high level test 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 e fusing the Sigma
126. is damaged and or has been used more than five times contact the Technical Assistance Center for a new set of shipping materials 2 22 Installation Chapter 3 Operation This chapter includes instructions for operating the ELx808 and its on board software and contains the following sections SN A E 3 2 The KE Pda eret teer ete e a e e er ee ias 3 2 The Startup Screen edet retener e tete eie tade tinent estes baee ides 3 3 The Main Menu Sereen teet dde t ee de UR ENT nes ERA TE caes 3 3 DefimingASSays ai e e e d de dee fea Mater cel 3 5 Selecting an ASSay 5 Net ed et e eet a quede tei reete ID 3 5 ASSAY INdIIe ese notet eet ca ie E Pe M TI deS 3 6 Defining the Method Map Formula and Curve sse 3 7 METHOD tia 3 7 MA E 3 16 FORMULAS di adios 3 31 CURE de e e a fae pelea A Mr tds 3 43 PANAS Sii A AA AA Aid 3 47 Reading Microplate ue rne HE RE RC EVER PESE 3 50 Printing Reports and Assay Lists ccccccssecssecsseceteceeceeceeceeeeeeeeseeeeeeeeeaeecaeessees 3 54 Rel adi oe td edi odit ate Eae e leet e toe es 3 55 Editing Standard Outliers sse 3 55 Pont Resultszs 5uete ttr tetro A ee rete viec 3 56 NUES HE E Et DTE 3 57 NSS AY stone PREND date ERR DEA QU Pe e eere ERR cee 3 57 Introduction The Keypad 3 2 The ELx808 features a 25 pad keypad and a 2 x 24 character LCD display as the user interface The reader s bidirectional serial port allows computer cont
127. istical calculations e Curve Fitting with 4 parameter cubic quadratic linear 2 p cubic spline or point to point e Formula calculations for more complex mathematical operations e Ability to define controls and positive and negative cutoffs Specimen Preparation Samples should be obtained treated and stored following instructions and recommendations contained in the package kit Registration Card Once the Microplate Reader 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 ELx808 Operator s Manual xiii Warranty Bio Tek Instruments Warranty Policy Bio Tek grants Limited Warranties that are applied to hardware and software products manufactured by Bio Tek Instruments Inc Bio Tek This Limited 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 damage and or bodily injury or death that may result from the use or misuse of any instrument software by the purchaser his employees agents or customers other than the express warranty contained herein Under no circumstances shall Bio Tek be liable to the purchaser or any other person for any direct incidental or consequential damages whether
128. lect ASSAY SELECT ASSAY NUMBER NAM BE HBS AG e Use the keyboard to type the assay number or the OPTION key to cycle through a list of available assays Press ENTER to enter the assay and begin printing the map and other assay parameters List e Select REPORT at the Main Menu and then select LIST The entire list of assays stored in the ELx808 s memory will be sent to the printer ELx808 Operator s Manual 3 57 3 58 Operation Chapter 4 Performance Verification Periodic Maintenance and IQ PQ OQ Tests This chapter discusses the tasks and procedures necessary for verifying and qualifying instrument performance initially and on an ongoing basis A convenient Recommended Test and Maintenance Schedule arranges tasks into Operation and Performance Qualification and Maintenance categories This chapter contains the following sections Recommendations for Achieving Optimum Performance sse 4 2 Installation Qualification IQ nennen 4 3 Recommended Test and Maintenance Schedule OQ PQ PM sss 4 4 Qualification Procedures e ea A tete tete EU ripe o I ped 4 5 Initiating Tests Via the Utility Option enne 4 6 SYSTEM System Test 2 rettet teret ert iet fe er da 4 7 CHKSUM Checksum Test aia 4 9 CALPLATE Absorbance Plate Test 4 9 Absorbance Plate Test ed ete eere eo eire ede nna 4 10 Requirements itat E t ade e eats eens 4 11 Entering the Test Plat
129. line of the display and the available options appear on the bottom GENERATION MANUAL MANUAL Automatic Plate Map Generation Select AUTO to instruct the software to automatically generate a plate map after the blanks controls standards and or samples have been defined Manual Plate Map Generation Select MANUAL to indicate that the well assignments will be performed manually by the user at Define and or Read time e Press SOFT KEY 1 for automatic sample plate map generation The display will update to reflect the selection e Press SOFT KEY 2 for MANUAL plate map generation The display updates to reflect the selection e Press ENTER to save the selection and move to the next screen Note Press SHIFT CLEAR to clear any previously defined manual map ELx808 Operator s Manual 3 17 MAPPING DIRECTION Use this option to select how the wells are mapped on the plate Any previously defined Mapping Direction appears on the top line of the display the available options appear on the second line MAPPING DIRECTION DOWN DOWN ACROSS e Press SOFT KEY 1 to map DOWN the column e Press SOFT KEY 2 to map ACROSS the row e Press ENTER to save the selection and move to the next screen REPLICATION DIRECTION This option allows you to 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 botto
130. ll as the ability to program variables for use within Transformation formulas 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 or call POS NEG or EQUIV e TRANSformation formulas are applied to the raw data in preparation for further data reduction and or curve fit calculation e VALadation 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 e Note GENERAL formulas are not used in the ELx808 open assays C T FORMULA TYPE FF TRANS A L Cc T FORMUTLA TYPE RAL TRAN S VAR e Press SOFT KEY 1 to select CUTOFF Formula e Press SOFT KEY 2 to select TRANSFORMATION Formula e Press SOFT KEY 3 to select ASSAY VALIDATION Formula e Press MORE to access additional formula types e Press SOFT KEY 3 to select TRANS VAR e Press ENTER to save the selected formula type and advance to the next screen 3 32 Operation VALIDATION TYPE SELECTION If you selected VAL this option allows you to choose which Validation Formula type Control Assay or Blank Validation formulas to enter for the assay SELECT VALIDATION TYPE CONTROL ASSAY BLANK Press SOF
131. losed as the shroud is being lifted to prevent the door from moving 2 4 Installation e Ensure that all locations on the filter wheel contain either a filter or a blank Each location on the filter wheel must be occupied for the reader to operate properly Take a moment to record the filter values in each location e g 405 nm in position 1 filters must match the filter locations mapped in the reader s software filter D Important Keep track of all filter locations The physical location of the table The filter wheel must have no empty locations all locations must be filled with either a filter or a blank plug Install all filters with the arrow denoting the light direction pointing downward e The filter wheel mount is located in the rear left corner inside the reader The filter wheel is held in place by a magnet on the filter wheel motor hub To install the filter wheel l Line up the registration notch on the hub and the corresponding peg on the filter wheel Apply firm pressure to attach the filter wheel to the hub The peg must be engaged in the notch for proper installation Ensure that the filter wheel is positioned flat against the hub and that it rotates freely Lower the top shroud back into position and remove tape if used to steady the plate access door Reinstall the seven screws removed from the perimeter of the shroud reader s ability to withstand electrostatic discharges and electromagnetic
132. m R E P DIRECTION DOWN ACROSS e Press SOFT KEY 1 to map the replicates DOWN the column following the direction of the map listing e Press SOFT KEY 2 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 advance Examples of mapping and replication directions are shown on the next page 3 18 Operation EXAMPLES OF MAPPING amp REPLICATION DIRECTIONS Map Direction DOWN Rep Direction DOWN E Uie a AA A E ES EL 8 0 do o A J SIDL STIDS SPP j j J LB smi ems supe Y p h p c ps SID ROS SMA o 3 spe ego o BO EO dh A GA De STD E cp qs p cap cimo cd Eo Ae A if ESPOSAS RNC He po 3 AE E o cupo pe xo f PET IBN aiii eee LG SIDA sel l dq oc spl boue Ae EY p AA vw ug dp o uw T Map Direction ACROSS Rep Direction ACROSS p umo MAS E cou 1s te CUT ideo B NC NC SMP SMP SMP SMP SMP se Er a a Map Direction DOWN Rep Direction ACROSS pS 2 ee ES E A pe 1 ee ee 4 ASA SOT poe cocer TE A MNAE HA B Sis aepo al cg xp ly Ae dno US Ae a EXC US cupo AL o cup p quz n LO c sm sm p p qo d o odo p E SIDS stos o oe AMA SHE SE A AA 7 RES PEA EA AM E AE AE AE AH ECER ENG EEE 2 Ac AS cro ato 24i 3 0x pH TUSMPpeMPCE in uc wwe cw c uo
133. nalog PCB or faulty internal grounding Displayed Error Potential Cause ERROR 0900 Read time dark value out of range Error 900 indicates that the dark current value taken during the current read is significantly different than the same reading taken during the power up self check PROBABLE CAUSE e Read chamber door has opened or closed relative to when the unit was powered ON Close door and reboot reader to correct e Electrical noise penetrating the measurement chamber The bottom and top shrouds are part of the electrical shielding Check that the shrouds are installed and properly fastened e Internal electronic noise caused by a faulty Analog PCB or faulty internal grounding C 6 Error Codes Displayed Error Potential Cause ERROR 0A00 0A01 0A05 Read time air blank out of range Error A00 indicates that the blank full signal reading taken during the current read has changed significantly from the same reading taken during the power up self check PROBABLE CAUSE e Lamp has failed since power up Check that the lamp is lit and replace if necessary e Analog PCB failure Displayed Error Potential Cause Error C00 indicates that the time allotted for the instrument to make a valid connection to a printer has expired PROBABLE CAUSE e Printer out of paper not connected or not on line Appendix C C 7 C 8 Displayed Error ERROR A900 ERROR 0D00 ERROR 0E00 0E01 0E05 ERROR 0F00 0F8
134. nd more To read a plate using KCjunior l 2 B 4 Click Read Plate from KCjunior s main screen The Read Plate dialog will appear If desired enter a Results ID and a Plate Description and then click Read Plate The Protocol Definition dialog will appear Select a Read Method Type of Endpoint Kinetic 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 e The plate will be read and then the raw data results will display in KCjunior Print the raw data by selecting Plate Print Results e To analyze or manipulate results a protocol should be defined Refer to KCjunior s Help system or User s Guide for instructions Computer Control Controlling the Reader With KC4 Follow these steps to establish communication between the ELx808TM and KC4 1 Appendix B Turn off the computer and the reader Connect the appropriate serial cable PN 75053 between the two machines Turn on 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 ELx808 reader model Click the Port but
135. ned as 999 0x03E7 If the instrument does not have incubation or an incubation error has been detected 0x0000 will be returned as temperature data An incubator temperature error will then be flagged and returned with the standard status response string Otherwise the current temperature averaged over four thermal zones is returned as defined above The reader will accept and process the Get Temperature command at any time including during a read cycle The temperature response will NOT interrupt a plate data response stream however If a Get Temperature command is received during a data transmission it will not be processed until the transmission has completed i e after the lt Z gt checksum combination has been sent GET PLATE BARCODE s This command returns the barcode for the microplate currently in the instrument carrier host valid limits response Is no arguments lt ACK NAK gt barcode 32 status string 5 The barcode is returned as a stream of 32 characters If the barcode itself is less than 32 characters it is padded with nulls 0x00 up through the 32 character Otherwise the barcode is truncated to 32 characters and sent with no null terminator If no barcode is detected after two passes of the microplate carrier in front of the scanner the null terminated string NR is returned followed by the remaining 29 nulls A barcode error is flagged and returned with the standard status response string
136. ng 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 Appendix E Automatic Microplate Plate Washer This is recommended however plate 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 E 3 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 ofthe cutoff and the ANA result of the Negative and Positive Controls should be in the ranges specified on the QC Certificate 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 Valu
137. o many scans are chosen the reader may be reading the sides of the well The reader will read the chosen number of points across the well and report the calculated area under the curve ELx808 Operator s Manual 3 15 MAP The MAP Definition screen allows you to edit or specify the following options in the assay e Automatic or Manual Map Generation e Mapping Direction e Replication Direction e Blank Map Selection e Blanking Constant e Number of Blanks e Location of Blanks e Number of Standards e Number of Standard Replicates e Averaging of Standards e Concentration and Location of Standards e Number of Controls e Control Type Definition e Number of Control Replicates e Control Location e Number of Samples e Number of Sample Replicates e Averaging of Samples e Sample Location reader model The valid range of the number of standards is 0 12 The valid range of valid replicate counts for standards is 1 8 The valid range of the number of controls is 0 8 The range of valid replicate counts for controls and samples is 1 12 D Note Some of the following screens may not appear depending on the 3 16 Operation FORMULA CURV E e Atthe DEFINE Options screen press SOFT KEY 2 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 The currently selected value appears on the top
138. o select the reader and define the communications parameters choose the following setup parameters Reader ELx808 EL808 or ELx808I EL808l or ELx808U EL808U or ELx808UI EL808UI Com Port Select the COM port used for the RS 232 serial cable connection Baud Rate 9600 change the baud rate on the reader if necessary Data Bits 8 Parity None Stop Bits 2 EOT Character Keep the default number Click the Test Communications button to attempt to establish communications with the reader using the currently defined communication parameters If a Serial Write Error dialog box is displayed an incorrect Com Port may have been selected Select a different port and then repeat this step If the test passes click OK to save the settings and close the dialog box If the test fails follow the directions provided by KCjunior then click Test Communications again See also the Problems section on the next page B 3 Problems If KCjunior M 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 a
139. ociocniocnnocnnonnconconnncon nooo nonononnocononos 2 6 Reconfiguring or Replacing Fuses essen 2 8 Connect Po Wei tia 2 10 Install the Printe seision rne ttp D a a PEN ET ERFURT Ce Nd 2 11 Pr da datada cda 2 12 Sc EE 2 12 Parallel Port Pin Definition sessssssesesseeeeeeenee eene 2 12 Turn on the Reader and Run a System Test sss 2 13 Check Adjust the Reader s Filter Table sese 2 14 Configure Utility Op Sic ai e rete oreet teret aes 2 15 SETUP Options O reste ais 2 15 OUTPUT Options ite ete tete e Ee E ert eer ias 2 16 RBPOR LIyDe entrate titi ts ese Pieter 2 17 READ Options choise A V TEE 2 18 Read Speed zie eee eO Re eee e e gue NN 2 18 Set Up the Serial Port for Communication with Other Devices 2 19 Attaching the Serial Cable sse enne enne 2 19 Setting Communications Parameters ccccccssecssecsteceteceeceeeeeeeeseeeeeeeeeeeennes 2 19 Serial Port Pinout Description sessssssssseeeee eene nnne 2 21 After Installation and Setup Verify Performance cooonoocnnonnnonnnonanononannnon nono nonnncononos 2 22 Before Repackaging the Instrument ooooonocnnncnioninonononoconoconaconoconoconnnnono nan roannoan cnn 2 22 Unpack and Inspect the Instrument If the shipping box has been damaged inspect the instrument for visible dents and scratches as you unpack it If the reader is damaged notify the carrier and you
140. omputer Control sees eeeseeeeuesuuee B 1 Appendix C Error Codes ce rror ettet iori Eee Sero eie p enne tb Peta iia iiini C 1 Appendix D Report Format ueeeueeeueee sees eeeeeeeeeeeeeereeeee D 1 Appendix E Instructions for Programming a New Assay E 1 Appendix F Instructions for Adjusting the Line Input Voltage Range F 1 ELx808 Operator s Manual xix Preface Chapter 1 Introduction This chapter introduces the ELx808TM Absorbance Microplate Reader and describes its hardware and software features Also included is contact information if technical assistance is needed This chapter contains the following sections Introducing the ELx808 Absorbance Microplate Reader sssssss 1 2 Quality Controla ao 1 2 Hardware Feat tes etes iro t ee E arena iS oasis 1 3 Software Features tiet ER bie er tee TE Sa can iii da 1 3 Specifications 1 5 dedit Ph E E itae te ient ideas 1 4 Standard Model err t tei ente ie eie HERE leere E 1 4 Ultraviolet UV Model sese nennen nnns 1 5 Incubator Model 3 EE tease T E n EAT E UR TEUER a 1 5 Package Contents inier ee I f a oi ep en eet eder sees 1 6 Optional Accessories 5 55 dM ep E Ehe duele EE ani 1 6 Technical Support cete ee dencdecascoudacssnessedaaaediaesstesdedassardacesnecsedes 1 7 PHONE Supportin OA 1 7 Electronic Comm
141. ontact the Bio Tek Service Department deionized or distilled water and wipe the keypad Dry immediately with a Do not soak the keypad this will cause damage Moisten a clean cloth with clean dry cloth Important Turn off and unplug the instrument for all decontamination and cleaning operations Procedure 1 Turn off and unplug the instrument 2 Moisten a clean cotton cloth with water or with water and the mild detergent Do not soak the cloth 3 Whpe the plate carrier the inside of the plate carrier door and all exposed surfaces of the instrument 4 Ifdetergent was used wipe all surfaces with a cloth moistened with water 5 Usea clean dry cloth to dry all wet surfaces 6 Reassemble the instrument as necessary Appendix A A 5 Decontamination Appendix B Computer Control The ELx808 can be controlled either from the reader s front panel or from a computer connected to the reader via the computer s serial port This appendix describes the features of computer control and explains the information necessary to program the computer to control the reader This appendix contains the following sections OVerVIG Wet RT B 2 Controlling the Reader With KCjunior M 0 0 eccccccecseeetecsteceeceeceeeeeseeeeeeeeeeennes B 3 Problems ccd dedi AA LII LIT B 4 Getting Started With KCjunior ooooooocnoonnnonooonnonnconncon ccoo a a a e aa B 4 Controlling the Reader With KC4 eese B 5 Pr PEE a A B 6 Getting
142. or reliability of software or equipment that is not supplied by Bio Tek or its affiliated dealers ELx808 Operator s Manual iii Revision History Rev Date A 1 96 B 5 96 C 1 97 D 2 98 E 8 98 F 3 99 G 7 99 H 5 00 I 7 02 J 11 03 Changes First Release Revised reader specifications Added Scanning method to software Added Panel and Reuse of Standard Curve and revised serial port pin out description Changed Chapter 4 Performance Verification to reflect changes to the Universal Test Plate Added Liquid Tests 1 and 2 Changed Appendix C Error Codes Added printer information Updated Appendix B Computer Control Changed European address Corrected printer compatibility Corrected the sequence of steps for processing a curve fit method Added comment about blanking P Down and P Across being inactive in this version of software Clarified the need for at least one sample to be defined on a plate Added scanning computer control commands Updated Chapter 4 Performance Verification to include a Liquid Test 3 to verify 340 nm instruments Corrected the voltage range for Range 2 in the Introduction Corrected Incubation Temperature Control range to 6 above ambient Changed Note on page 4 7 dealing with air readings during Self Test Clarified Liquid Test 1 Changed Table 4 1 to include additional maintenance items Reworked Liquid Test 2 Updated contact information pages iii 1 8 1 9 2 8 2 20 and 4 13
143. ormation is printed on the side of the bottle Commercial bleach is typically 10 0 NaClO if this is the case prepare a 1 20 dilution Household bleach is typically 5 0 NaClO if this is the case prepare a 1 10 dilution Moisten a cloth with the bleach solution or alcohol Do not soak the cloth Wipe down the carrier and all exposed instrument surfaces with the bleach solution gt 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 gt Wipe the plate carrier the inside of the carrier door and all exposed surfaces of the instrument Wait 20 minutes Moisten a cloth with deionized or distilled water and wipe all surfaces of the instrument that have been cleaned with the bleach solution or alcohol Use a clean dry cloth to dry all wet surfaces Reassemble the instrument as necessary Discard the used gloves and cloths using a Biohazard trash bag and an approved Biohazard container Decontamination Routine Cleaning Procedure Purpose Exposed surfaces may be cleaned not decontaminated with a cloth moistened not soaked with water or water and a mild detergent Tools and Supplies e Mild detergent e Deionized or distilled water e Clean cotton cloths not allow the cleaning solution to run into the interior of the instrument If this Do not immerse the instrument spray it with liquid or use a wet cloth Do happens c
144. programmed programmed in the reader s memory e The assay s name and number are displayed on the screen ELECT ASSAY NUMBER AME HBS AGl e Press ENTER to select the assay and advance to the NAME screen You may change the default assay name to a more descriptive one see Assay Name on the next page e Press ENTER to advance to the EDIT ASSAY NAME screen ELx808 Operator s Manual 3 5 Assay Name 3 6 At the ASSAY NAME screen edit the name assigned to the Assay The assay name can be up to 16 characters IT gt HBS AG 1 In addition Press the ALPHA and NUMERIC keys to update the assay name The cursor is positioned at the first editable field Press the OPTION key to sequentially advance the character positioned above the cursor The characters will cycle through the alphabet A Z with a space character following Z Press CLEAR to clear the assay name from the display Use the LEFT and RIGHT ARROWS to move the cursor to the previous or next editable field The cursor will wrap around the edit field Press SOFT KEYS 1 2 3 and 4 when using alphanumeric characters on the display above the soft key in the assay name Press the MAIN MENU key to return to the Main Menu screen Press PREVIOUS SCREEN to save the contents of the display and return to the previous screen Press ENTER to save the contents of the display and advance to the next screen Operation Defining the Method Map Formula
145. ptions in Chapter 2 Installation In addition an Assay List Assay Definition Map and Result can be printed by choosing Report from the Main Menu screen Bio Tek Instruments Assay ASSAY 01 Date 07 23 96 Lot Time 03 05 35AM Operator Wavelength 405 Temp Plate ID SONTA COMMENTS OUT indicates an out of range value was read 1 2 3 4 5 7 8 9 10 41 12 A eT e E drum a ae a CALL POS POS POS POS POS POS POS POS POS POS POS Calc0D 0 182 0 072 0 073 0 077 0 071 0 075 0 073 0 074 0 073 0 075 0 072 0 076 Wel STD1 SMPS SMP13 SHP21 SMP29 SMP37 SMP45 SHPD3 SMP61 SMP69 SMP77 SMP85 SS ERES a irc RETE ES CALL POS POS POS POS POS POS POS POS POS POS POS Calc0D 0 529 0 076 0 084 0 085 0 096 0 073 0 073 0 069 0 072 0 070 0 070 0 076 Well STD SMP6 SMP14 SHP22 SMP30 SMP38 SMP46 SMP54 SMP62 SMP70 SMP78 SMP86 RSLT 0 483 CALL POS NEG NEG NEG POS POS POS POS POS POS POS Calc0D 0 721 0 070 0 351 0 443 0 376 0 076 0 074 0 077 0 071 0 076 0 081 0 066 Well STD3 SMP7 SMP15 SHP23 SMP31 SMP39 SMP47 SNP55 SMP63 SMP71 SHP79 SMP87 RSLT 0 686 0 294 0 391 0 321 CALL POS NEG NEG NEG POS POS POS POS POS POS NEG Calc0D 0 749 0 081 OUT OUT QUT 0 078 0 074 0 079 0 074 0 072 0 076 0 683 Well 5TD4 SMP8 SMP16 SHP24 SMP32 SMP40 SMP48 SMP56 SMP G4 SMPT SHP80 SNP88 RSLT 0 715 0 646 _ _ Qq _ _ _ _ CALL POS POS POS POS POS POS POS POS POS POS POS POS CalcOD 0 068 0 070 0 093 0 101 0 081 0 078 0 078 0 074 0 074 0 071 0 074 0 072 Well SMP1 SHPI S
146. quipment e Disposal Notice x Preface Safety Symbols The following warning and informational symbols may be found in various locations on the ELx808 The warnings are given here in English French German Spanish and Italian Only qualified personnel who recognize shock hazards and are familiar with the safety precautions should use this instrument Read the manual carefully before operating this instrument Ei e ELx808 Operator s Manual Consult instructions for use Consulter la notice d emploi Gebrauchsanweisung beachten Consultar las instrucciones de uso Consultare le istruzioni per l 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 Alternating current Courant alternatif Wechselstrom Corriente alterna Corrente alternata Direct current Courant continu Gleichstrom Corriente continua Corrente continua Both direct and alternating current Courant continu et courant alternatif Gleich und Wechselstrom Corriente continua y corriente alterna Corrente continua e corrente alternata Earth ground terminal Borne de terre Erde Betriebserde Borne de tierra Terra di funzionamento Protective conductor terminal Borne de terre de protection Schutzleiteranschluss Borne de tierra de protecci n Terra di protezione xi 2 Pe PO On Supply Marche
147. r equal to 2 e Usethe LEFT and RIGHT ARROW keys to move to the next or previous numeric entry fields e Press ENTER to save the value and advance to the next screen KINETIC NUMBER OF READS Use this screen to enter the number of kinetic reads TOTAL NUMBER OF KINETIC READS e Use the NUMERIC keys to enter the number of reads required The range is 2 to 40 reads e Press ENTER to save the entry and advance to the next screen ELx808 Operator s Manual 3 11 KINETIC DURATION Use this screen to enter in hours minutes and seconds the duration of the kinetic reaction KINETIC DURATION e Use the NUMERIC keys to enter the time duration in hours minutes and seconds The maximum duration time is 80 hours e Use the LEFT and RIGHT ARROW keys to move between entry fields e Press ENTER to save the entry and advance to the next screen SHAKE MODE SELECTION Use this screen to enter the shake mode for a kinetic or endpoint assay SHAKE BEFORE EVERY REA D FIRST EVER Y NONE e Press SOFT KEY 1 to select shaking for the first read only or endpoint read e Press SOFT KEY 2 to select shaking for every read e Press SOFT KEY 3 to select no shaking e Press ENTER to save the selection and advance to the next screen e SCREEN key to save the selection and return to a previous screen Operation SHAKE TIME Use this screen to enter the desired shake interval Continuous appears on the display when a kin
148. r manufacturer s representative Keep the shipping cartons and the packing material for the carrier s inspection The manufacturer will arrange for the repair or replacement of your instrument immediately before the shipping related claim is settled Important Save all packaging materials If you need to ship the reader to Bio Tek for repair or replacement you must use the original packing Other forms of commercially available packing are not recommended and can void the warranty Refer to the figure on the next page when unpacking the reader Carefully open the top of the box and remove any accessories These include a power cord a filter wheel in a padded envelope and an Operator s Manual Remove the end caps from the reader Lift the reader out of the box and place it on a level surface Remove the reader from the plastic bag Remove the filter wheel from the shipping envelope Place all shipping material back into the shipping box for reuse if the instrument needs to be shipped again Installation Figure 2 1 Unpacking the ELx808 Select the Operating Environment For best operation install Bio Tek s ELx808 on a level surface in an area where ambient temperatures between 18 C 65 F and 40 C 104 F can be maintained The reader is sensitive to extreme environmental conditions Conditions to avoid are e Excessive humidity Condensation directly on the sensitive electronic circuits can c
149. rate that is 1f the Accuracy portion of this test passed To further prove linearity you can perform a regression analysis on the Test Plate OD values in a program such as Microsoft Excel as follows 1l Launch Excel 2 Open a spreadsheet and label one column Assigned and the next column Observed 3 Enter the Assigned OD data for each glass filter in the first column from the data sheet provided with the Test Plate Analyze one wavelength at a time 4 Enter the Observed OD values for the same glass filters in the adjacent column Performance Verification and IQ OQ PQ Tests 5 Under Tools select Data Analysis and then Regression Use the Regression Input box to enter the Assigned values as the Input Y Range and the Observed OD as the Input X Range Note Ifthe Data Analysis command is not available on the Tools menu you may need to install the Analysis ToolPak in Microsoft Excel Consult Microsoft Excel Help for assistance 6 Click OK and the Summary Output sheet will be displayed An R Square value of at least 0 99 is expected Repeated Test Failures If any portion of the test continues to fail contact Bio Tek s Technical Assistance Center refer to Chapter 1 for contact information Please have a copy of the test report and the reader s serial number available when you call ELx808 Operator s Manual 4 15 Empty Carrier Test The Empty Carrier Test confirms the ELx808 s re
150. rator s Manual PN 7341000 and Warranty Registration Card Dust cover PN 7342066 Parallel cable PN 71072 Serial cable PN 75053 Optional Accessories 1 6 Additional filters please inquire for availability 340 to 630 nm PN 3100XXX XXX is wavelength in nanometers 640 to 900 nm PN 3404XXX XXX is wavelength in nanometers Replacement lamp assembly PN 3400508 Filter wheel plug PN 3122037 Absorbance Test Plate PN 7260522 Service Manual for all instrument models PN 7341004 Introduction Technical Support Bio Tek s ELx808 is backed by a superior support staff If the ELx808 ever fails to work perfectly please contact Bio Tek s Technical Assistance Center You can call write fax or E mail your questions and concerns to Bio Tek using the contact information provided below Whichever method of contact you choose please be prepared to provide the following information Product name and serial number The specific steps that produce your problem Any error codes displayed on the screen A daytime phone number Your name and company information A fax number and or an 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 RMA number and be sure to repackage the reader properly see Chapter 2 page 2 2 Phone Support Telephone the Technical Assistance Center TAC between 8 30 AM and 5 30 PM Eastern Stand
151. rint the results The Installation Qualification is now complete ELx808 Operator s Manual 4 3 Recommended Test and Maintenance Schedule OQ PQ PM The schedule shown below 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 Qualification procedures be performed more or less frequently than shown below Table 4 1 Recommended Test and Maintenance Schedule Operational Performance Maintenance Qualification Qualification E th E Initially and Annually Monthly pe oa Ed System Test p 4 7 Y Checksum Test p 4 9 Y Absorbance Plate Test p 4 10 Y Empty Carrier Test Y p 4 16 Liquid Test 1 p 4 20 or Y Every three months Liquid Test 2 p 4 22 Liquid Test 3 p 4 25 optional for 340 nm Clean Light Bulb J Contacts p 4 28 Inspect Clean Wavelength Filters Y p 4 29 Lubricate Robotic Components p 4 30 As needed after RIS ATS tmp us approximately 500 hours Regarding Liquid Tests 1 and 2 If you have an Absorbance Test Plate run Liquid Test 1 If you do not have an Absorbance Test Plate run Liquid Test 2 Liquid Test 3 is optional it is provided for sites requiring verification at wavelengths lower than those attainable with the Absorbance Test Plate 4 4 Performance Verification and IQ OQ PQ Tests Qu
152. rol of the instrument and provides the means for downloading additional assay protocols to the instrument Te 84 9 CLEAR ENTER 4 5e 6r la 2s 3c OO O shit STOP READ Figure 3 1 Keypad Use the four soft keys located directly below the display to select options presented on the display Use the labeled keys to enter information move around the reader s menu structure specify the assay number and so on CLEAR Clear reset the current value on the reader s LCD display ENTER Advance the display to the next screen Main Menu Return the display to the Main Menu screen Previous Screen Return the display to the previous screen Options Cycle through the available options presented on the display STOP Halt a read in progress READ Select an assay and read a plate Operation The Startup Screen The ELx808 performs a System Test when powered on displaying the Startup screen until initialization is complete During this period the keyboard is inactive If System Test fails the reader will beep and display an error code Refer to Appendix C Error Codes to interpret this code For further information please call the Technical Assistance Center see Chapter 1 page 1 7 for contact information The Main Menu Screen Once the system is initialized the Main Menu Screen is displayed This screen will vary slightly if the instrument has the Incubation option The keyboard s four
153. rosoft Excel Consult Microsoft Excel Help for assistance Expected Results Since it is somewhat difficult to achieve high pipetting accuracy when conducting linear dilutions an R Square value of at least 0 99 is considered adequate ELx808 Operator s Manual 4 27 Periodic Maintenance for the ELx808 Cleaning the Lamp Contacts 4 28 Over time an oxide coating forms on the electrical contacts on the reader s lamp This coating can reduce lamp output the contacts should be cleaned at least once every three months A Warning Turn off and unplug the ELx808 before cleaning the lamp contacts To clean the lamp contacts l 2 3 10 Notes If the reader is on turn it off and let the lamp cool for at least ten minutes Temporarily tape the reader s plate access door shut Using your fingertips locate the seven securing screws around the sides and front where the reader s top shroud meets the base Remove the screws using a flat blade screwdriver Tip Bring the reader to the edge of the work surface to access the screws without having to turn the reader upside down Carefully raise the front of the shroud and tip it toward the back of the instrument The lamp is located in the left rear quadrant inside the reader Unplug the lamp connector from the circuit board Note Do not touch the glass lens in the lamp bracket The lamp bracket is attached to the side of the instrument with two screws which
154. s been selected Any previously defined value is displayed 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 the ENTER key to save the value and move to the next screen NUMBER OF STANDARDS Use this option to enter the number of standards that will be used in the assay Any previously defined value will be displayed on the screen If the number of standards is altered the number of replicates for the standard automatically defaults 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 2 P 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 save the value and move to the next screen ELx808 Operator s Manual 3 23 NUMBER OF STANDARD REPLICATES This option allows you to enter the number of replicates per standard 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 m
155. s physical defects in the filters Displayed Error Potential Cause ERROR 0700 Reader failed noise test Error 700 indicates significant variations in background electronic noise were detected when blocking the light and increasing the Gain to maximum PROBABLE CAUSE e Electrical noise penetrating the measurement chamber The bottom and top shrouds are part of the electrical shielding Check that the shrouds are installed and properly fastened e Ambient light leak Check that the plate chamber door is closed to keep out ambient light e Missing or loose filter allowing unintended lamp light into the measurement chamber Check filter wheel for empty positions e Internal electronic noise caused by a faulty Analog PCB or faulty internal grounding Appendix C C 5 Displayed Error Potential Cause ERROR 0800 Reader failed offset test Error 800 indicates background electronic signal detected is outside of acceptable limits at maximum gain when blocking the light PROBABLE CAUSE e Electrical noise penetrating the measurement chamber The bottom and top shrouds are part of the electrical shielding Check that the shrouds are installed and properly fastened e Ambient light leak Check that the plate chamber door is closed to keep out ambient light e Missing or loose filter allowing unintended lamp light into the measurement chamber Check filter wheel for empty positions e Internal electronic noise caused by a faulty A
156. s 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 autoantibody 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 E 2 Instructions for Programming a New Assay Materials 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 Ill Wash Buffer 20x Concentrate 1 bottle containing 50 ml of a concentrated buffer for washing the wells Type Ill 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 serum Ready to use ANA Cutoff Control 1 bottle containing 1 8 ml of diluted stabilized serum Ready to use ANA Negative Control 1 bottle containing 1 8 ml of diluted stabilized serum Ready to use ANA Conjugate 1 bottle containing 12 ml of peroxidase labeled antibody to IgG Colored red ready to use TMB Substrate 1 bottle containi
157. s to enter the greyzone percentage e The valid entry range is from 00 to 9996 An entry of 00 indicates no greyzone although a sample equal to the cutoff value will still receive the EQUIV call e See POS NEG Calls below for information on how calls are assigned e Press the CLEAR key to clear the displayed value e Press the ENTER key to save the displayed value and advance POSITIVE NEGATIVE CALLS FOR CUTOFF After the greyzone is defined calls for the sample wells POSitive NEGative EQUIVocal must be defined SAMPLEOCUTOFFEF 0 5 amp NEG e Select POS or NEG to select the call that will be assigned to samples greater than the cutoff value plus the greyzone e If for example POS is selected 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 MP gt CUTOFF CUTOFF GREYZONE S POS SMP gt CUTOFF CUTOFF GREYZONE S NEG MP CUTOFF CUTOFF GREYZONE 3 38 Operation 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 e For this example NC x the mean of the NC wells equals 1 000 OD e The cutoff formula is NC x 0 5 e The greyzone is 00 e POS is selected for SAMP gt CUTOFF 00 e Calls
158. s used for classifying samples During data reduction results are evaluated against the cutoff value with an optional greyzone and each well is assigned a call POS positive NEG negative or EQUIV equivocal e One cutoff formula may be defined per assay e If Transformation Formulas are defined cutoffs are based on the transformed results Refer to FORMULA on page 3 31 for the order in which formulas are processed e Acutoff formula can consist of a simple numeric value 1 500 a well identifier PC or PC x to represent the mean or a formula combining the two NC x 0 050 e A greyzone around the cutoff value can be defined to indicate equivocal or indeterminate results e Do not use the lt or gt mathematical symbols in a cutoff formula e 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 After selecting an assay page 3 5 define the Cutoff formula as shown below FORMULA CURVE ORMULA TYPE RANS A L e After the FORMULA 1 prompt disappears enter the formula as described in Formula Entry on page 3 34 Refer to the examples on page 3 39 ELx808 Operator s Manual 3 37 GREYZONE ENTRY The greyzone is a definable area around the cutoff value Samples falling within an area defined by the greyzone ex 5 0 of the cutoff value could be considered equivocal EQUIV ENTER GREYZONE e Use the NUMERIC key
159. say definition table is specified This command has no effect on the read parameters for a wellset read Appendix B B 17 SET COLUMN RANGE This command selects a range of adjacent columns numbered on plate or strip carrier to be read host valid limits response M lt ACK NAK gt data bytes 1 first column 1 24 2 last column 1 24 status string 5 By default the instrument is initialized to perform full plate reads and automatically reverts to this default when a new assay definition table is loaded either from the front panel or by computer control The well range is also reset when the Set Geometry command is sent Once this command is invoked the reader will use the ELx format for data response until a new assay definition table is specified This command has no effect on the read parameters for a wellset read GET MINIMUM KINETIC INTERVAL This command provides a means for the user to determine the minimum kinetic read interval as dictated by the current assay definition table well range and even baud rate selected host valid limits response no arguments lt ACK NAK gt kinetic interval 2 status string 5 The reader returns the minimum kinetic interval as a 2 byte integer value not ASCII so low byte first Minimum kinetic intervals are not available for wellset reads B 18 Computer Control SELF TEST gt This command causes the reader to perform a syst
160. se 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 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 The inclination of the meniscus can cause loss of 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 Although the effect of ambient light is mathematically quantified and subtracted from each absorbance reading the illumination of the instrument by strong ambient light should be avoided If interference from ambient light is suspected read a microplate of high density solutions under the suspect conditions and again with all ambient light blocked in a dark room for example then compare results The blocked results will appear as an increase in optical density readings if l
161. se of reader error data previously received during the current read sequence should be discarded 312 Format This format is selected by default at reader powerup and every time a new assay definition is loaded either via serial control V command or by the user from the front panel The reader always returns data as an 8 x 12 matrix along with the assay name and extra characters There are no checksums or final terminator ELx Format If any of the following occurs the ELx format is selected and remains enabled until a new definition is loaded either via serial control V command or by the user from the front panel 1 Multi wavelength mode is selected in the assay definition 2 Arange of rows is set even if for all rows see Set Row Range 3 Arange of columns is set even if for all columns see Set Column Range The reader returns data in a matrix corresponding to the rows and columns selected for up to three wavelengths If dual wavelength subtraction is selected only one plate shall be returned not available in the multi wavelength mode Checksum calculation starts with the first byte AFTER the wavelength start code up through and including the Z data terminator code at the end of each wavelength s plate data The checksum is transmitted as an integer data byte not ASCII The final terminator indicates that the data stream is complete for the corresponding Read Plate command No more data shall be transmitte
162. sing 1000 100 and 10 wl Multichannel Pipette Recommended for dispensing 100 ul volumes of conjugate substrate and stop solution Glass Plastic Tubes For sample dilution E 9 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 QC Certificate e The curve shape should be similar to the standard curve shown on the QC Certificate 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 CV for each duplicate OD is less than 15 0 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 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
163. steps alloc free error lt assay num data flash write timed out data flash readback didn t match write test lt chip gt code flash write timed out memory allocation heap corrupted no errors detected read function aborted lt motor gt didn t find opto sensor transition motor didn t find saturation transition motor failed positional verify A D signal saturated test type filter gain out of range reader channel failed noise test reader channel failed offset test read time channel filter dark out of range QA00 0B00 0C00 0D00 QE00 OF00 1000 1100 1200 1300 1500 1600 1700 1800 1900 read time channel lt filter gt air blank out invalid lt assay number gt printer timed out failed calibration checksum test wavelength not found in table lt read filter gt channel lt filter gt signal out of range necessary configuration data missing failed configuration checksum test necessary calibration data missing motor not homed successfully incubator failure error code lt zone s gt computer control assay definition error interval too short for selected options too many kinetic intervals malloc failed B 25 STORE CURVE ERR 1A00 store curve failure GET CURVE ERR 1B00 get curve failure
164. t is always facing the ON OFF switch when installed in the power input module The text matching the desired voltage range should be on the edge of the switchboard which goes into the power input module as the arrow on the board indicates The white plastic indicator will need to rotate around so that it fits into the correct groove on the switchboard e Once the switchboard is reinstalled the white plastic indicator peg should line up with the fuse holder voltage indicator holes If the peg does not fit in the hole which indicates the voltage intended the switchboard is not installed correctly Do not power up the instrument until the voltage input range to be used is indicated correctly by the peg Adjusting the Fuse Configuration or Replacing the Fuse Both USA and European fuses are installed in the reader s power input fuse module The reader s fuses are configured at the factory prior to shipping Use the following procedure if you need to change the fuse configuration or replace fuses A failed fuse is usually an indication of another problem which a new fuse is not likely to fix Contact Technical Services if the fuse replacement fails to rectify the problem e Usea small flat blade screwdriver to remove the fuse module from the power input module The fuse module has two fuse configurations gt USA A fused hot 1 5 slo blo amp PN 46024 gt Europe Both hot and neutral fused 0 63 slo blo 5 x 20 mm amps PN 46038 e To replac
165. t of light on both wavelengths e Press ENTER to save the selection and advance to the next screen ELx808 Operator s Manual 3 9 MEAS SELECTION The MEAS selection screen allows you to select the filter s for the assay The currently selected filter appears on the top line of the display and the available options appear on the bottom e Press SOFT KEYS 1 2 3 and 4 to select the filter option displayed above the soft key The display updates to reflect the selection e Press the right arrow key to select the Reference Filter e Press ENTER to move to the next screen NUMBER OF KINETIC READS KINETIC DURATION SELECTION This menu allows you to either select the total number of kinetic reads or the length of time the assay will run kinetic duration Any previously defined value is shown on the top line of the display and the options on the second KINETIC TOTAL READS TOTAL READS DURATION e Press SOFT KEY 1 to select the total reads option e Press SOFT KEY 3 to select the duration option e Press ENTER to save the selection and advance to the next screen Operation KINETIC INTERVAL Use this screen to enter the interval of time in hours minutes and seconds between each kinetic read KINETIC INTERVAL e Use the NUMERIC keys to enter the time duration Valid ranges are 0 1 hours 0 59 minutes and 0 59 seconds The number of Reads Duration Interval must be less than or equal to 40 and more than o
166. t of the expected range review the other set for the same channel pair Thus if A1 and H12 are not within range of each other review the compliance of H1 to A12 A2 to H11 and H2 to A11 This will confirm that there is a problem in one of the eight read channels or indicate that the result of one set of wells was in error If any two sets of well values for a channel pair are out of the allowed accuracy range there may be contamination on one of the lenses If the four corner wells are within the repeatability range the reader is also in alignment Performance Verification and IQ OQ PQ Tests Liquid Test 3 Note A 96 well flat bottom microplate is required for this test Corning Costar 3590 is recommended Use a new microplate any fingerprints or scratches may cause variations in the turnaround reading REQUIRED MATERIALS e 340 nm filter installed in the reader e New 96 well flat bottom microplate Corning Costar 3590 recommended e Calibrated hand pipette s e Beakers and graduated cylinder e Precision balance with readability of 0 01 g e Buffer Solution A or B SOLUTION A 10X CONCENTRATE PBS e Deionized water e Ingredients shown in Table 4 5 e p NADH Powder B Nicotinamide Adenine Dinucleotide Reduced Form Sigma bulk catalog number N 8129 or pre weighed 10 mg vials Sigma number 340 110 Note Store the B NADH powder according to the guidelines on its packaging 1 Prepare the stock buffer solution us
167. te in that liquid in the wells has a meniscus while the Test Plate s neutral density filters do not The optics characteristics may differ in these two cases alerting you to different types of problems The Absorbance Test Plate will indicate the absolute amount of light absorbed which will effectively test the linearity of the electronics The liquid tests will help detect optical defects such as dirt on the lenses or other contamination that can contribute to errant readings Liquid Test 1 tests the alignment accuracy repeatability and channel to channel variability of the reader making evident any problems with the optics of the system If you have the Absorbance Test Plate you will only need to run Liquid Test 1 for routine testing If you do not have an Absorbance Test Plate test the linearity repeatability and alignment of the reader by preparing a series of solutions of varying absorbances as described in Liquid Test 2 Liquid Test 3 is an optional test offered for sites which must have proof of linearity at wavelengths lower than those attainable with the Absorbance Test Plate This test is considered optional because the reader has good front end linearity throughout its wavelength range 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 4 3 The purpose of the
168. the door roller in the closed position Press the OPEN soft key Press the UP soft key to raise the door to the desired height Press the DOWN soft key 1f you want to adjust the door downward Important Do not open the door to its maximum open position Press the Main Menu key Press the CLOSE soft key This will close the door The door should rest closed on the top cover ELx808 Operator s Manual 4 31 Replacing the Lamp The reader s lamp should operate without replacement for about 500 hours The intensity of the lamp will slowly drop over time until the reader s run time self check detects a low signal and warns the user with a displayed error code The bulb should be replaced at this time PN 3400508 The figure below provides a detailed view of access to the lamp Socket To main ys circuit board gre e Spring clip 4 32 Bulb Perimeter screws 7 Flat head screws 2 Figure 4 6 Location of the lamp in the ELx808 Performance Verification and IQ OQ PQ Tests immediately after the unit has been turned off so allow at least 10 minutes O Warning Turn off and unplug the instrument The lamp will be hot cool down time before attempting to replace the lamp 1 After unplugging the instrument remove the seven perimeter slotted black screws from the front left and right sides of the instrument 2 Lift the top shroud up to expose the interior of the instrument The lamp assembly is loc
169. the next screen or use the MAIN MENU and PREVIOUS SCREEN keys to move backward through the menu structure Operation EDIT STANDARD OUTLIERS This screen allows you to select which method None or Manual will be used to edit Standard Outlier values After the standard curve has been calculated one or more standards can be excluded from the recalculation of the curve Any previously defined edit method is displayed EDIT STD OUTLIERS MANUAL NONE MANUAL e Press SOFT KEY 1 or 2 to select the edit option displayed above the soft key The display updates to reflect your selection e Select NONE to suppress the Edit Standard Outliers capability for this assay e Choose MANUAL to enable the capability gt IfAVERAGE STANDARDS is set to NO the individual standard replicates are available for editing If set to YES the standard groups are available for editing gt After the assay is run and reports are generated press REPORT from the Main Menu Press RESULT select the assay and then press ENTER The EDIT STD OUTLIERS YES NO prompt will appear See Editing Standard Outliers later in this chapter for further instructions e Use the ENTER key to save the selection and advance to the next screen or use the MAIN MENU and PREVIOUS SCREEN keys to move backward through the menu structure ELx808 Operator s Manual 3 45 AXIS SELECTION This screen allows you to select the X and Y Axis Type Any previously defined axis type w
170. thin Quick Read You can access the Quick Read assay when READ is selected from the main menu D Important On some readers Assay 01 has been designed to allow for quick The Quick Read assay DEFINE settings are shown below They cannot be edited except where noted METHOD Single Wavelength 405 nm editable MAP 96 well plate geometry Blank on Air Automap Map starting location A1 Samples only no blanks standard or controls Sample count prompted at runtime can be turned off in UTIL READ options 3 4 Operation 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 O Note Appendix E provides provides two examples of assay kit instructions Defining Assays The Main Menu option DEFINE allows you to customize previously defined assays stored in the reader s memory e From the Main Menu press the soft key beneath the DEFINE menu option to advance to the SELECT ASSAY NUMBER screen Selecting an Assay At the SELECT ASSAY NUMBER screen e Use the NUMERIC keys to enter the number of predefined Assay Definition files stored in the reader s memory or use the OPTION key to advance one assay at a time The cursor is positioned at the first editable field and advances automatically The numeric range depends on the number of assays 1 55 or more if custom
171. tion 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 Within any given formula type the order of processing is the order in which the formulas are entered VALIDATION FORMULA EXAMPLES Blank Validation An assay protocol states that every blank well on a plate should have an OD of less than 0 050 The formula is entered on the reader as a Blank Validation Formula BLK lt 0 050 Negative Control Validation An assay protocol states that every Negative Control well must have an OD of less than or equal to 0 100 The formula is entered as a Control Validation Formula NC 0 100 Positive Control Validation An assay protocol states that every Positive Control well must have an OD higher than 1 000 but less than 2 500 Two Control Validation Formulas can be entered PC gt 1 000 AND PC lt 2 500 Or one formula can be used if the formula is 24 characters or less PC gt 1 000ANDPC lt 2 500 Assay Validation An assay protocol states that in order for an assay to be valid the mean of the Negative Control well ODs must be less than 0 100 The Assay Validation formula that should be entered NC x lt 0 100 the map identifier NC x indicates the mean of the NCs From the assay DEFINE Menu press the arrow corresponding to FORMULA ELx808 Operator s Manual 3 31 Formula Type The ELx808 supports three types of formulas as we
172. 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 Liquids Avoid spilling liquids on the instrument fluid seepage into internal components creates a potential shock hazard Wipe up all spills immediately Do not operate the instrument if internal components have been exposed to fluid 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 Software Quality Control The operator must follow the manufacturer s assay package insert when modifying software parameters and establishing read 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 analyz
173. ton and subsequent Setup button to define the following communications parameters Port Select the COM port used for the RS 232 serial cable connection Baud Rate 2400 default 4800 or 9600 must match the baud rate on the reader Data Bits 8 Parity None 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 on the next page B 5 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 l 2 B 6 Select Data New Plate If prompted to select a protocol select Emp
174. tthe SELECT UTILITY OPTION screen press the soft key beneath OUTPUT to set report output preferences REPORT OUTPUT BOTH PRINT COMPUTER BOTH e Any previously defined selection appears on the top line of the display Press the soft key beneath the desired option PRINT COMPUTER BOTH to change the output device e Press ENTER to advance to the SELECT PRINTER menu screen If COMPUTER is pressed all results will be transferred directly to the computer screen via the RS232 serial port SELECIT PRINTER EPSON H P Compatible Printers BTI readers support printers using either HP s PCL3 language such as the HP DeskJet series or Epson s LQ language For the latest list of compatible printers call Bio Tek s Technical Assistance Center see Chapter 1 for contact information or visit our Web site www biotek com 2 16 Installation REPORT Type Note See Appendix D for examples of Reports e Atthe SELECT PRINTER screen press ENTER to advance to the REPORT TYPE menu screen MATRIX MATRIX e The current selection appears on the top line of the display Press the soft key beneath the desired option COLUMN MATRIX BOTH to change the Report Type The display will automatically update with the new type e Press ENTER to advance to the SAMPLES ON COLUMN REPORT screen e Select YES to print samples on the column report or NO to omit them e Press ENTER to advance to the PRINT CURVE FIT screen
175. ty 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 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 e The plate will be read and then the raw data results will display in KC4 e To analyze manipulate or print results protocol parameters should be defined Refer to KC4 s Help system or User s Guide for instructions Computer Control Controlling the Reader Using Serial Protocol At baud rates of 1200 2400 and 9600 the ELx808 is capable of sending and receiving data through its serial port RS 232C The baud rate used for transmission is held in non volatile memory and can be changed by the user Other serial port parameters Parity None Data Bits 8 and Stop Bits 2 are fixed and cannot be changed The reader s RS 232C serial port is configured as a DTE that is the unit is wired to resemble a modem Data is received on Pin 3 the RX Pin and transmitted on Pin 2 the TX pin See Chapter 2 Set Up the Serial Port for Communications with Other Devices
176. ty of the OD readings is confirmed by default if the wavelength readings are accurate Empty Carrier Test Confirms the ELx808 s read capabilities at the 10096 light level Liquid Tests Verify that the reader will perform to specification with liquid samples Conduct these tests again monthly quarterly as outlined in the schedule as a part of Performance Qualification The following pages describe how to run these tests via the reader s keypad They can also be run via KCjunior and KC4 Refer to the appropriate user guides for instructions ELx808 Operator s Manual 4 5 Initiating Tests Via the Utility Option 4 6 01 30PM 1 DEFINE REPO Select SYSTEM to run the SYSTEM TEST Select CHKSUM to run the CHECKSUM TEST Select CALPLATE to run the ABSORBANCE PLATE TEST Note Before running this test ensure that the Test Plate values are entered See Entering the Absorbance Plate Specifications on page 4 11 for more information Performance Verification and IQ OQ PQ Tests SYSTEM System Test The System Test conducts a series of tests at each wavelength defined in the filter table to confirm adequate light levels low electronic noise adequate photodiode sensitivity overall system cleanliness and 1f equipped proper function of the incubator The testing is designed to verify that the ELx808TM will give in specification performance for each set wavelength over the specified OD range The reader automatically
177. uld be set to 0 GET WAVELENGTH TABLE W This command uploads the wavelength setup table from the reader host valid limits response ww no arguments lt ACK NAK gt reader response protocol for each wavelength configured six wavelength 000 900 comma terminator status string 5 B 16 Computer Control HALT X gt This command causes any read in progress to be halted host valid limits response x no arguments none see below The following events occur when this command is invoked 1 The read process is halted and all axes are returned to their home positions the plate is moved back outside where it can be accessed 2 The reader transmits a lt DLE gt character to the computer when the above process is completed 3 No more data is transmitted by the read or scan in progress A final status string is transmitted in the ELx Status Mode SET ROW RANGE r This command selects a range of adjacent rows lettered on plate or strip carrier to be read host valid limits response Y lt ACK NAK gt data bytes 1 first row 1 16 2 last row 1 16 status string 5 By default the instrument is initialized to perform full plate reads and automatically reverts to this default when a new assay definition table is loaded either from the front panel or by computer control Once this command is invoked the reader will use the ELx format for data response until a new as
178. ultiplied 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 save the value and move to the next screen AVERAGE STANDARDS The Average Standards option allows you to select whether or not to average the Replicates of a Standard This average is used to calculate the standard curve instead of using the individual replicate of each standard Note If the replicate selection is 1 this option is not available AVERAGE STANDARDS YES NO e Press SOFT KEY 1 to select YES average the replicates The top line of the display updates to reflect the selection e Press SOFT KEY 2 to select NO do not average the replicates The top line of the display updates to reflect the selection e Press the ENTER key to save the selection and advance to the next screen 3 24 Operation STANDARD CONCENTRATION The Standard Concentration field allows you to enter the predicted or expected concentration value for each standard group Any previously defined value is displayed If Manual Map Generation is selected the replicate locations must also be defined LOCATION RE P 4 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 Valid well locations for the defined geometry are listed below e
179. ultiply by 100 Enter the formula OD 2 100 This formula will be applied to the ODs of all samples standards controls and blanks that are present on the plate map Operation TRANSFORMATION SCOPE VARIABLE For more complex transformations a Transformation Scope Variable TVar can be defined for use with a transformation formula This variable defines the scope of the transformation whether to apply the transformation to just the samples SMP or to all wells defined on the plate OD From the assay Define Menu press the arrow corresponding to Formula FORMULA CURVE This will bring you to a screen prompting you to Select Formula Type Press MORE at this screen SELECT FORMULA TYPE CUTOFF TRANS VAL The options displayed now include TRANS VAR Press VAR at this screen SELECT FORMULA TYPE GENERAL TRANS VAR The following screen will appear asking you to choose the scope of this transformation SCOPE VARIABLE S MP O D If SMP is chosen the transformation formula will be applied only to the samples defined in the plate map If OD is chosen the formula definition screen will appear 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 writing the transformation formula ELx808 Operator s Manual 3 41 Example An
180. unication ssssesseseeeee eee eene 1 7 Bax SUDDOELt eet erret ee teh E Pr t ee ex Pr voe de e erre Dea 1 8 Written Communications taen enr eere 1 8 Shipping Address eit i E ee EE en ere EORR 1 8 Introducing the ELx808 Absorbance Microplate Reader Bio Tek s ELx808 is an eight channel reader assay system The reader can serve as a standalone system or can be controlled via PC based software Designed to automatically perform endpoint and kinetic analysis the reader can measure the optical density of solutions in 96 well microplates between 380 nm and 900 nm The UV option measures down to 340 nm The reader features superior optical specifications with an extended dynamic range of up to 4 000 absorbance units The instrument s onboard processor 2 x 24 character LCD screen and membrane keys allow easy definition and management of assay protocols templates formulas and data Results can be output in a printed report format or exported for use in a variety of ELISA based data manipulation applications Bio Tek s ELx808 may be configured with all or selected options for optimum performance e U model instruments are capable of reading plates between 340 nm and 900 nm wavelengths e model instruments have a four zone incubation chamber that controls temperature up to 50 C Quality Control 1 2 It is considered good laboratory practice to run laboratory samples according to instructions and specific re
181. ure Utility Options x icit tc nep e ii 2 15 SETUP Obpltioris 2 Etre pe a Pe oes tee Lodo Eee ced pede eed 2 15 OUTPUT Options escrit cte deve ekle dete e tue a Ee ded Te dene nes 2 16 REPORT Type ede ede Dae ede ec ede ed eee Te ede e ie tet 2 17 ISEAD Optlons aaa 2 18 REAd Speed ie obe d eee e eet metet tem ert 2 18 Set Up the Serial Port for Communication with Other Devices seenen 2 19 Attaching the Serial Cable itio ertet irent tre rere decet eee nianie 2 19 Setting Communication Parameters ssssssssseee eene nenne 2 19 Serial Port Pinout Description serari annen edita canica n 2 21 After Installation and Setup Verify Performance sssssssee eee 2 22 Before Repackaging the Instrument sse eene eene 2 22 Chapter 3 Operation die dd 3 1 WtrOCUCtON ita OR Bi Gees eie ee Ge rea 3 2 The Kepa pe adebetee 3 2 The Startup Sereen ioo ER deret a ia eden s 3 3 The Main Menu Second de Ee ER Ue teo dede d een 3 3 Defining ASSAYS etariko Em 3 5 Selecting AMAS iii A ae 3 5 ASA NAME cea oput ode dre eee eet etre 3 6 Defining the Method Map Formula and Curve ssssssssseeeeeneeen 3 7 METHOD 2 5 A icut estt ve i dc Me P ses ged oeste Set Saccus 3 7 ghi sm 3 8 Delay in First Read een reete tegi unes enne E eR Re SEA UY Sed 3 8 Incubation Temperature
182. vised 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 Where protective gloves when performing the decontamination procedure Decontamination Tools and Supplies e Sodium Hypochlorite NaCIO or bleach e 70 isopropyl alcohol alternative to bleach e Deionized or distilled water e Safety glasses e Surgical mask e Protective gloves e Lab coat e Biohazard trash bags e 125 ml beakers e Clean cotton cloths Decontamination 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 Appendix A A 3 Turn off and unplug the instrument Prepare an aqueous solution of 0 5 sodium hypochlorite 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 inf
183. wes m n Computer Control The following section describes the supported computer control command set STORE PLATE CARRIER A This command causes the plate carrier to move inside the instrument host valid limits response A no arguments lt ACK gt status string 5 4 READ WELL SET d This command causes the reader to return data collected at the center of each of the indicated wells in the order specified The parameters for the actual data collection are taken from the currently loaded assay definition table see Set Assay Definition host valid limits response e lt ACK NAK gt data bytes 1 2 well 1 row 01 16 3 4 well 1 column 01 24 5 8 well 2 9 12 well 3 13 16 well 4 17 20 well 5 21 24 well 6 25 28 well 7 29 32 well 8 33 position in read series 0 single read 1 first read in series 2 next read in series 3 final read in series lt ETX gt lt ACK gt if data valid Appendix B B 9 reader response protocol initial status string 5 for each kinetic interval one if endpoint read for each wavelength data start code 1 lt CR gt for each of eight wells comma separator 1 4 sign 1 or data 4 1234 row terminator 2 lt CR gt lt LF gt temperature 3 if specified 000 999 wavelength terminator 1 lt Z gt checksum 1 0 255 final terminator 1 lt Z gt
184. 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 pre coated 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 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 Materials Materials Supplied e Instruction Leaflet Giving full assay details e QC Certificate Indicating the expected performance of the batch E 8 Instructions for Programming a New Assay 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
185. wording into an ELx808TM assay program We are providing the following programming examples using fictitious In Vitro Diagnostic IVD assays These instructions are not intended to promote the use of the ELx808 with these or any other specific IVD assays and they do not constitute a claim of IVD use in jurisdictions where there are additional regulatory requirements For clarity only the user menu choices from the reader screens are shown Refer to Chapter 3 for details D Important The kit instructions are provided so that users can see how it is properly according to their specific kit instructions and for verifying that the calculations are performed correctly If you are not familiar with programming formulas onboard the ELx808 refer to Formula Entry in Chapter 3 for further information D Important The ELx808 user is responsible for programming the reader This appendix contains the following sections Sample ANA Screen Enzyme Immunoassay Kit Transformation and Cutoff seen enne E 2 Programming the ANA Screen Enzyme Immunoassay Kit Transformation and Cutoff ooooocinocononaconoonnnonnnonccnn nono nonn nono noconono noc n enne E 5 Sample Anticardiolipin IgG Enzyme Immunoassay Kit Standard Curve and Cutotf n nen e Er Pr I ene eie E 8 Programming the Anticardiolipin IgG Enzyme Immunoassay Kit Standard Curve and Cutoff sssssssssseseseeeeeeen enne ennt E 12 Sample ANA S
186. y DI Water to bring volume to 1000 ml Preparation of Stock Solution 1 Weigh out 0 092 gram of FD amp C No 5 yellow dye powder into a weigh boat 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 KF YN Make up to 1 liter with DI water cap and shake well This should create a solution with an absorbance of about 2 000 when using 200 ul in a flat bottom microwell The OD value result will be proportional to the volume in the well and the amount of FD amp C No 5 dye used You can use a larger or smaller well volume or add more dye or water to adjust the solution Note that too small a well volume may result in increased pipetting related errors 4 18 Performance Verification and IQ OQ PQ Tests SOLUTION B Required Materials e Bio Tek QC Check Solution No 1 P N 7120779 25 ml 7120782 125 ml e Deionized water e 5 ml Class A Volumetric Pipette e 100 ml Volumetric Flask Preparation of Stock Solution 1 Pipette a 5 ml aliquot of Bio Tek QC Check Solution No 1 into a 100 ml volumetric flask 2 Make up to 100 ml with DI water cap and shake well gt This should create a solution with an absorbance of about 2 000 when using 200 ul in a flat bottom microwell The OD value result will be proportional to the volume in the well and the amount of QC Check Solution No 1 used You can use a larger or smaller well volume or add more Check Solution or

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