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Bio-Tek ELx800 Operator'S Manual Download Page 73

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45

Important:

On some Universal Microplate Readers,

Assay 01

has been designed to allow for quick and

simple programming. It appears as “_Quick Read” on
the display. Most of the options available in Assays
2-55, and described in this section, are unavailable for
programming within Quick Read. You can quickly
access the Quick Read assay by selecting

READ

from

the main menu.

The Quick Read assay default

DEFINE

settings are shown below, and cannot be edited,

except where noted.

Method

•

Single Wavelength 405 nm (editable)

Map

•

96-well plate geometry

•

Blank on Air

•

Automap

•

Map starting location A1

•

Samples only (no blanks, standards or controls)

•

Sample count prompted at runtime (can be turned off in UTIL|READ options)

ELx800 Operator’s Manual

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Summary of Contents for ELx800

Page 1: ...ELx800 Absorbance Microplate Reader Operator s Manual...

Page 2: ......

Page 3: ...ed software installed Not all features of the software discussed in this Operator s Manual are available on custom instruments Please contact Bio Tek s Technical Assistance Center at 800 242 4685 if y...

Page 4: ......

Page 5: ...ELx800 Absorbance Microplate Reader Operator s Manual October 2004 2004 Part Number 7331000 Revision N Bio Tek Instruments Inc...

Page 6: ...4 KCjunior and EL800 are trademarks of Bio Tek Instruments Inc Microsoft Windows and the Windows logo are registered trademarks of Microsoft Corporation in the United States and other countries All ot...

Page 7: ...ontrol xv Repackaging and Shipping xv Warnings xvi General xvi Readers xvi Hazards and Precautions xvii Hazards xvii Precautions xviii Directive 89 336 EEC Electromagnetic Compatibility xix Directive...

Page 8: ...itten Communication 11 Shipping Address 11 Chapter 2 Installation 13 Unpacking and Inspecting the ELx800 14 Unpacking the Instrument and Its Accessories 15 Setting Up the ELx800 19 Operating Environme...

Page 9: ...ine 46 Programming Note 47 Selecting an Assay to Define 48 Editing the Assay Name 48 Define Method Map Formula and Curve 49 Defining METHOD 49 Defining MAP 53 Defining FORMULA 69 Defining CURVE 81 Rea...

Page 10: ...Entering the Absorbance Plate Specifications 113 Test Failures 115 Liquid Testing 116 Stock Solution Formulation 117 Liquid Test 1 119 Liquid Test 2 121 Liquid Test 3 for UV Models Only 125 Chapter 5...

Page 11: ...olling the Reader With KCjunior or KC4 149 Overview 150 Controlling the Reader With KCjunior 151 Problems 152 Getting Started With KCjunior 152 Controlling the Reader With KC4 153 Problems 154 Getting...

Page 12: ...5 Quality Control and Results 176 Expected Values 176 Programming the ANA Screen Enzyme Immunoassay Kit 177 Sample Human Anticardiolipin IgG Enzyme Immunoassay Kit 179 Intended Use 179 Background 179...

Page 13: ...he Unit in the Box 36 Figure 14 Keyboard 40 Figure 15 Options Available from the Main Menu 44 Figure 16 Sample Output for the System Test Sheet 1 of 2 108 Figure 17 Sample Output for the System Test A...

Page 14: ...Column Report Without Samples 167 Figure 35 Panel Report 168 Figure 36 Assay Detail Report Sheet 1 of 2 169 Figure 37 Assay Detail Report Sheet 2 of 2 170 List of Tables Table 1 Recommended Test Sched...

Page 15: ...2 655 4040 outside the U S Fax 802 655 7941 E Mail customercare biotek com Service TAC Phone 800 242 4685 toll free in the U S 802 655 4740 outside the U S Fax 802 655 3399 E Mail tac biotek com Europ...

Page 16: ...the potential for bodily harm and tells you how to avoid the problem Caution A Caution indicates potential damage to the instrument and tells you how to avoid the problem DEFINE Text in COURIER font...

Page 17: ...L800 model comparison table in Appendix F I 2 99 Incorporated Manual Updates Changed European addresses J 9 99 Changed printer compatible description Changed the number of samples necessary if there a...

Page 18: ...page 1 8 Changed callout in Figure 2 2 from Place unit in bag to Unit in plastic bag Chapter 2 page 2 2 Added KC4 startup information to Appendix E Removed references to ELx800UV and EL800UV from Appe...

Page 19: ...ce to run laboratory samples according to instructions and specific recommendations included in the assay package insert for the test to be conducted Failure to conduct Quality Control checks could re...

Page 20: ...values may be affected by extraneous particles such as dust in the microplate wells A clean work area is necessary to ensure accurate readings When operated in a safe environment according to the inst...

Page 21: ...wear safety glasses and appropriate protective equipment such as chemically resistant rubber gloves and apron Warning Liquids Avoid spilling liquids on the instrument fluid seepage into internal compo...

Page 22: ...ely affected if temperatures fluctuate above or below this range Storage temperature limits are broader Caution Sodium Hypochlorite Do not expose any part of the instrument to the recommended diluted...

Page 23: ...age EN 61010 1 2001 Safety requirement for electrical equipment for measurement control and laboratory use Part 1 General requirements Directive 2002 96 EC Waste Electrical and Electronic Equipment Di...

Page 24: ...epartment 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 Depar...

Page 25: ...leich 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 Prote...

Page 26: ...surface Attention surface chaude Warnen hei e Oberfl che Precauci n superficie caliente Attenzione superficie calda Consult instructions for use Consulter la notice d emploi Gebrauchsanweisung beacht...

Page 27: ...OID if the Product has been repaired or altered by persons not authorized by Bio Tek or if the Product has had the serial number altered effaced or removed This Warranty is VOID if the Product has not...

Page 28: ...n 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 i...

Page 29: ...ware features Also included is contact information if technical assistance is needed Introducing the ELx800 Automated Microplate Reader 2 Hardware Features 3 Software Features 3 Package Contents 4 Opt...

Page 30: ...0 nm UV instruments have an extended range from 340 nm to 750 nm Kinetic analysis can be performed using computer control e g via KC4 or KCjunior running on a host PC Assay definitions consisting of p...

Page 31: ...lude 384 well and 60 72 96 well Terasaki microplates External 24 volt power supply which runs on 100 to 240 V 10 0 50 to 60 Hz One serial COM port 25 pin male connector One parallel port 25 pin female...

Page 32: ...340 nm filter Operator s Manual PN 7331000 and Registration Card Printer Cable PN 71072 Dust Cover PN 7332040 RS 232 Serial Cable PN 75053 Unpacking instructions PN 7271003 packing instructions PN 727...

Page 33: ...and 96 well Terasaki plates for ELx800NB and ELx800RNB PN 7330531 Absorbance Test Plate PN 7260522 Epson LX 300 printer or equivalent 110V only PN 97145 Sheet feeder for Epson LX 300 PN 97144 HP Desk...

Page 34: ...D at 405 nm 3 0 from 2 000 OD to 3 000 OD 450 nm Repeatability STD 0 5 0 005 OD from 0 000 to 2 000 OD 405 nm The following specifications apply to 96 well flat or round bottom plates single wavelengt...

Page 35: ...y 1 0 from 0 000 to 2 000 OD 405 nm 3 0 from 2 000 to 3 000 OD 450 nm Repeatability STD 0 5 0 005 OD from 0 000 to 2 000 OD 405 nm Optical specifications for the 340 to 400 nm range 50 second read in...

Page 36: ...width interference filters User accessible filter wheel Up to 5 filters may be installed on the instrument at one time Filters supplied 405 nm 450 nm 490 nm and 630 nm The following specifications app...

Page 37: ...a well to obtain the specified performance is 80 microliters The specifications apply to the dual wavelength mode of read only Absorbance Measurement Range 0 000 to 3 000 OD Normal Read Mode Dual wave...

Page 38: ...or codes displayed on the screen descriptions of error codes are available in Chapter 7 A daytime phone number Your name and company information A fax number and or e mail address if available If you...

Page 39: ...hnical Assistance Center 802 655 3399 Written Communication If you prefer you may write a letter with your comments and send it to Bio Tek Instruments Inc Technical Assistance Center P O Box 998 Highl...

Page 40: ...12 Chapter 1 Introduction Bio Tek Instruments Inc...

Page 41: ...t and Its Accessories 15 Setting Up the ELx800 19 Operating Environment 19 Electrical Connections 19 Power Up and System Test 20 ELx800 Main Menu 20 Configuring Global Default Options 21 Connecting a...

Page 42: ...amage during shipping Inspect the shipping box packaging instrument and accessories for signs of damage If the reader is damaged notify the carrier and your manufacturer s representative Keep the ship...

Page 43: ...ent and Its Accessories 1 Carefully open the top of the box and remove the power supply shelf Figure 1 2 Remove the top foam end caps manual and Declaration of Conformity Figure 1 Removing the power s...

Page 44: ...remove the foam end caps Figure 2 Place the reader on a level surface and remove the instrument from the plastic bag Figure 2 Removing the foam end caps Right end cap Unit inside 26 x 32 2 Mil poly b...

Page 45: ...level surface 5 Use the slotted screwdriver to remove the four screws from the top cover mounting Figure 3 and shipping straps from the bottom of the instrument Set aside these screws Figure 3 Removi...

Page 46: ...pping block Figure 4 Figure 4 Removing the shipping block Shipping block 8 Replace the top cover Turn the instrument upside down and replace the mounting screws 9 Remove the shipping straps from the t...

Page 47: ...roper operation of the unit The ELx800 has a universal 24 VDC 2 Amp power supply that functions from 100 to 240 V 10 0 50 to 60 Hz without external switching Never use a two prong plug adapter to conn...

Page 48: ...er up system test does not produce a printed results report To run the test manually and obtain a printout of the system test values start at the Main Menu and press UTIL TESTS SYSTEM See System Test...

Page 49: ...tional options include report format Column and or Matrix and whether or not to print standard curves READ Enable or disable read time prompting for Plate ID Sample ID and Sample Count Specify whether...

Page 50: ...rallel cable PN 71072 designed to connect the reader to a printer is provided with the ELx800 To attach a printer to the ELx800 1 Turn the reader off 2 Place the printer in a location adjacent to the...

Page 51: ...Setting Up the ELx800 23 Figure 6 Connectors for printer parallel computer serial and power supply Power supply jack Parallel port printer port Serial port ELx800 Operator s Manual...

Page 52: ...upgrades of the ELx800 software Appendix A contains information on required protocols for computer control of the reader Attaching the Cable 1 Power down the computer and the ELx800 2 Connect the app...

Page 53: ...T U P I N F O R M A T I O N D A T E T I M E F I L T E R M O R E E D I T S E T U P I N F O R M A T I O N R S 2 3 2 C A L P L A T E M O R E 4 The SELECT BAUD RATE screen will appear showing the current...

Page 54: ...portant Keep track of all filter locations The physical location of the filters must match the filter locations mapped in the reader s software filter table The filter wheel must have no empty locatio...

Page 55: ...can be lifted off 8 Remove the filters by turning the wheel upside down over a cloth The filters which are labeled with the wavelength and light direction arrow should easily slide out 9 When replaci...

Page 56: ...L S E L E C T U T I L I T Y O P T I O N T E S T S S E T U P O U T P U T R E A D 2 From the SELECT UTILITY OPTION menu press SETUP The Edit Setup Information screen appears on the display E D I T S E T...

Page 57: ...T H 4 0 5 When the last filter has been entered the software exits the filter routine and displays the following screen E D I T S E T U P I N F O R M A T I O N D A T E T I M E F I L T E R M O R E 6 Pr...

Page 58: ...original packing materials have been damaged or lost contact Bio Tek for replacement packing see Technical Assistance in Chapter 1 for contact information Warning If the reader has been exposed to pot...

Page 59: ...to Figures 8 through 13 when repackaging the ELx800 1 Move the carrier to the home position 2 Turn off the unit and unplug the power supply 3 Wrap the shipping straps around the carrier as shown in F...

Page 60: ...over and hook the shipping straps around the shaft Figure 9 5 Remove the four top cover mounting screws 6 Lift the instrument off the cover and turn it over Figure 9 Hooking the shipping straps aroun...

Page 61: ...ipping the ELx800 33 7 Install the shipping block Place the groove in the shipping block over the shaft and screw it down Figure 10 Figure 10 Installing the shipping block Shaft Shipping block ELx800...

Page 62: ...4 Chapter 2 Installation Bio Tek Instruments Inc 8 Turn the instrument over and install the top cover that was removed in step 6 Figure 11 Figure 11 Reinstalling the top cover Mounting screws Top cove...

Page 63: ...00 Operator s Manual 9 Turn the instrument right side up and put the end caps on the unit Figure 12 Figure 12 Reattaching the end caps Right end cap 26 x 32 2 Mil poly bag Left end cap 10 Place the un...

Page 64: ...lace the shelf into the notched out area in the end cap and the power supply into an 8 x 11 bubble bag Place the power supply on the shelf Figure 13 Figure 13 Placing the unit in the box Power supply...

Page 65: ...calibration cleaning periodic maintenance warranty work and or repair Make a note of any error messages displayed and their frequency 3 Provide Bio Tek with the name and contact information of a pers...

Page 66: ...38 Chapter 2 Installation Bio Tek Instruments Inc...

Page 67: ...Define 46 Define Method Map Formula and Curve 49 Defining METHOD 49 Defining MAP 53 Defining FORMULA 69 Defining CURVE 81 Reading a Microplate 88 Selecting an Assay to Run 89 Printing Reports 93 Edit...

Page 68: ...in Menu Previous Screen Options Main Menu Previous Screen Options 7 8 9 1 2 3 0 G H A B C 4 5 6 D E F Shift CLEAR ENTER STOP READ R E A D Y 9 4 5 A M 0 9 1 2 0 4 R E A D D E F I N E R E P O R T U T I...

Page 69: ...een settings and advances to the next screen in a series Pressing Previous Screen generally saves the current screen settings and returns control to the screen most previously viewed Press CLEAR to re...

Page 70: ...use 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 de...

Page 71: ...fails the System Test a chirp will sound and an error code will display Refer to System Self Test and Checksum Test in Chapter 4 for more information Refer to Chapter 6 Troubleshooting and Error Codes...

Page 72: ...reading and data reduction protocol You will be prompted to select an assay from an assay definition list REPORT option Allows printing of results and protocol descriptions You will be prompted for th...

Page 73: ...rogramming within Quick Read You can quickly access the Quick Read assay by selecting READ from the main menu The Quick Read assay default DEFINE settings are shown below and cannot be edited except w...

Page 74: ...ssay name and then press ENTER See Editing the Assay Name on page 48 for detailed instructions The DEFINE menu will appear D E F I N E M E T H O D M A P F O R M U L A C U R V E The following options a...

Page 75: ...Menu press DEFINE MAP and continue to press ENTER until ENTER NUMBER OF SAMPLES is displayed Enter at least one sample The software will lock up if zero 00 samples are selected E N T E R N U M B E R...

Page 76: ...reader s display MAIN MENU Returns the display to the Main Menu screen PREVIOUS SCREEN Returns the display to the previous screen ENTER Saves the current settings and advances to the next screen Editi...

Page 77: ...OD Specify the wavelength type wavelength filter s and plate geometry MAP Specify mapping information FORMULA Access the formula entry screens CURVE FIT Specify curve fit options Defining METHOD Defin...

Page 78: ...e final reported optical density is the difference between the two readings the delta OD Dual wavelength readings can significantly reduce optical interference caused by scratched or fingerprinted mic...

Page 79: ...4 well Nunc square 96 T Terasaki 72 T Terasaki 60 T Terasaki Used only on ELx800NB version The PLATE TYPE selection screen allows you to select the geometry of the plate that will be used for the assa...

Page 80: ...L A T E T Y P E 9 6 W E L L S 9 6 M M O R E Press MORE to cycle through the available options 6 6 well 2 x 3 96 96 well 8 x 12 12 12 well 3 x 4 96H 96 well Hellma Quartz 8 x 12 24 24 well 4 x 6 96M 96...

Page 81: ...Averaging of Standards Concentration and Location of Standards Number of Controls Control Type Definition Number of Control Replicates Control Location Number of Samples Number of Sample Replicates Sa...

Page 82: ...ards and or samples are assigned to specific locations on the plate M A P G E N E R A T I O N M A N U A L A U T O M A N U A L Select AUTOMATIC PLATE MAP GENERATION to instruct the software to automati...

Page 83: ...nd continue 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 a...

Page 84: ...3 4 5 6 7 8 9 10 11 12 A STD1 STD1 STD2 STD2 STD3 STD3 STD4 STD4 STD5 STD5 PC PC B NC NC SMP SMP SMP SMP SMP SMP C D E F G H Map Direction DOWN Rep Direction ACROSS 1 2 3 4 5 6 7 8 9 10 11 12 A STD1 S...

Page 85: ...G A T W E L L L O C A T I O N A 0 1 Use the numeric and alpha keys to enter a letter or number at the cursor location For any well location only the alpha keys are active for the first character and...

Page 86: ...on three screens B L A N K M A P F U L L A I R F U L L C O N S T M O R E B L A N K M A P F U L L R O W C O L U M N M O R E B L A N K M A P F U L L P A C R O S S P D O W N M O R E Select the BLANK MAP...

Page 87: ...nual mapping to position blanks controls standards and samples COLUMN enables a single blank well or an average of blank wells to be placed in each column Since the maximum number of blanks is 12 and...

Page 88: ...vances automatically Press CLEAR to clear the value on the display Press ENTER to continue Number of Blanks The NUMBER OF BLANKS field allows you to enter the number of blank wells in the assay This e...

Page 89: ...continue Number of Standards This option allows you to enter the number of standard groups that will be used in the assay Any previously defined value will be displayed on the screen Note If the numb...

Page 90: ...CLEAR to clear the value on the display Press ENTER to continue Average Standards The AVERAGE STANDARDS option allows you to select whether or not to average the replicates of each standard group Thi...

Page 91: ...ssing ENTER advances to the concentration value entry for the next standard If manual mapping is selected the location must be defined Pressing ENTER from the standard concentration entry moves the cu...

Page 92: ...o samples on a plate to be run in Assay 51 To reuse a standard curve you must first program an assay in positions 31 through 55 and then run the assay During the defining process you will be prompted...

Page 93: ...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 m...

Page 94: ...osing an identifier for CONTROL 1 press ENTER to choose the identifier for the next control Number of Control Replicates The NUMBER OF CONTROL REPLICATES entry screen is displayed if the number of con...

Page 95: ...t replicate or sample group Number of Samples The number of sample groups on the plate can be defined here and or it can be defined at run time if UTIL READ PROMPT FOR SAMPLE COUNT is set to YES See S...

Page 96: ...are automatically performs a check to ensure that the number of replicates multiplied by the number of samples does not exceed the number of undefined wells remaining on the plate Sample Location If M...

Page 97: ...T FORMULA TYPE screen will appear described on the next page Calculation Structure During data reduction formulas are processed in the order shown below The number of permitted formulas of each type i...

Page 98: ...ing data reduction results are evaluated against the cutoff formulas and each well is assigned a user specified label POS NEG or EQUIV TRANSformation formulas are applied to the raw data in preparatio...

Page 99: ...o press the appropriate SOFT KEY For example press the MATH SOFT KEY several times to see etc When the desired option appears press the right arrow key to select it and advance to the next editable fi...

Page 100: ...options depend on the formula type and the current plate map MAP options resemble BLK x mean of the blank wells NC 1 the first NC well or OD every well FUNCTION The following functions can be used in...

Page 101: ...ol replicates or control mean If the criteria are not met results are considered suspect and the message RESULTS INVALID BLANK OR CONTROL VALIDATION FAILED appears on results reports One blank validat...

Page 102: ...Screen key to move backward through the menu structure Assay Validation Formulas Assay Validation formula s establish a set of criteria used to determine whether or not an assay can be considered vali...

Page 103: ...defined per assay A transformation formula can be simple ex OD 2 100 to multiply all wells on the plate by 100 or more complex with the inclusion of a predefined Transformation Variable see TVAR belo...

Page 104: ...ill appear so that you can define a formula for use within the transformation formula Use the formula keys Math Other Map and Function to define the Transformation Variable TVAR Once the variable has...

Page 105: ...isplayed Choose TRANS Enter the formula OD TVAR TVAR 100 using the MATH OTHER MAP and FUNCTION keys TVAR is available as MAP option Example In the case of competitive reactions converting absorbance d...

Page 106: ...replicates or PC x to indicate the average of the Positive Control replicates or a formula combining the two NC x 0 050 A greyzone around the cutoff value can be defined to indicate equivocal or indet...

Page 107: ...s EQUIV SMP CUTOFF CUTOFF GREYZONE and SMP CUTOFF CUTOFF GREYZONE POS SMP CUTOFF CUTOFF GREYZONE NEG SMP CUTOFF CUTOFF GREYZONE Examples 1 The cutoff between negative and positive calls should be calc...

Page 108: ...tive All other samples should be considered equivocal For this example STD2 x the mean of the STD2 wells equals 2 000 OD The cutoff formula is simply STD2 x The greyzone is 10 POS is selected for SAMP...

Page 109: ...ters include Curve Fit Type Editing of Outliers Axis Identification Extrapolation of Unknowns Note These screens are displayed on the ELx800 in the order in which they appear in the assay If a closed...

Page 110: ...ch experimentally determined values are used for the responses at concentrations of zero and infinity Note Uses linear axis type only Cubic Spline C Spline curve fit A piecewise polynomial approximati...

Page 111: ...E M A N U A L Select NONE to suppress the EDIT STANDARD OUTLIERS capability for this assay Choose MANUAL to enable the capability If AVERAGE STANDARDS is set to NO the individual standard replicates a...

Page 112: ...samples outside of the absorbance range defined by the standards E X T R A P O L A T E U N K N O W N S Y E S Y E S N O Select YES to enable extrapolation otherwise select NO On the printed reports ext...

Page 113: ...defined for each assay will be used when the assay is selected for a Panel Panel assays cannot reuse standard curves The type and number of controls blanks standards and replicates in the assays chos...

Page 114: ...mapping directions for the predefined assays are overridden by the Panel s mapping directions If the assay includes replicates they will follow the Panel mapping direction After selecting the mapping...

Page 115: ...ll A01 The user wants to run 6 samples in Assay 1 Assay 1 now fills wells A01 through B03 The mapping for Assay 8 can begin in well B04 or any well other than A01 to B03 The reader will chirp if you t...

Page 116: ...BER screen will appear 2 Select an assay and then press ENTER See Selecting an Assay to Run on the next page for detailed instructions 3 If required enter the number of Samples Plate ID and or Sample...

Page 117: ...ding on preferences selected in UTIL READ whether or not the assay specifies manual mapping or if the assay was created or downloaded from Bio Tek s Extensions Define Reader Protocol software Prompts...

Page 118: ...ells remaining on the plate after any blank control or standard wells are mapped If you enter a value greater than the number of empty wells remaining on the plate the reader will chirp and automatica...

Page 119: ...tically increment each subsequent sample identification number by 1 The sample IDs will be assigned according to the plate map defined in the assay Prompts for Well Location If the assay specifies man...

Page 120: ...R E A D K E Y Before reading the plate ensure that the printer is connected turned on and full of paper Place the plate in the carrier and press the READ key to initiate the plate read After the read...

Page 121: ...E S U L T M A P A S S A Y L I S T Select RESULT to print an exact copy of results from the plate reading the 8 most recent sets of plate data are stored in memory The form in which the results are pr...

Page 122: ...t a plate and continue If a standard curve was generated and EDIT STANDARD OUTLIERS was set to manual in the assay definition the EDIT STD OUTLIERS prompt is presented otherwise the PRINT RESULTS prom...

Page 123: ...be chosen for exclusion E D I T S T D 1 X Y E S Y E S N O Select YES to exclude the group from curve fit calculations Select NO to retain the group Press ENTER to advance to the next group Each curve...

Page 124: ...S S A Y N U M B E R 0 1 N A M E H B S A G Press Options to cycle through the list of available assays or enter the number of the desired assay Press ENTER to print the report Assay Report The Assay Re...

Page 125: ...is positioned under the first editable field and advances automatically To change the date format press the soft key beneath MMDDYY or DDMMYY The display automatically updates to reflect the new forma...

Page 126: ...U P I N F O R M A T I O N D A T E T I M E F I L T E R M O R E 2 Select FILTER to display the wavelength for Filter 1 E N T E R F I L T E R 1 W A V E L E N G T H 4 0 5 3 Enter the correct wavelength fo...

Page 127: ...H P R I N T C O M P U T E R B O T H 2 The current output option is displayed on the top line Select PRINT to send reports directly to a printer COMPUTER to send data out through the serial port or BO...

Page 128: ...ix C for examples of Reports 7 Press ENTER to continue The SAMPLES IN COLUMN REPORT screen will appear S A M P L E S O N C O L R P T N O Y E S N O 8 Select YES to print results for all wells on the pl...

Page 129: ...E C O U N T Y E S Y E S N O R E A D I N R A P I D M O D E Y E S Y E S N O If selected at read time PLATE ID prompts for microplate identification SAMPLE ID prompts for identification for each sample g...

Page 130: ...102 Chapter 3 Operation Bio Tek Instruments Inc...

Page 131: ...ranges tasks into Installation Operation and Performance Qualification categories Recommendations for Achieving Optimum Performance 104 Recommended Test Schedule 105 Qualification Procedures 106 Syste...

Page 132: ...urements Inaccuracy in pipetting has a large effect on measurements especially if smaller volumes of liquid are used For best results use at least 100 l per well in a 96 well plate and 25 l in a 384 w...

Page 133: ...med more or less frequently than shown below Table 1 Recommended Test Schedule Installation Qualification Performance Qualification Monthly Performance Qualification Semiannually Operational Qualifica...

Page 134: ...the instrument or if regulatory requirements dictate that Performance Qualification Testing is necessary you should perform the following tests After the initial confirmation of operation you should p...

Page 135: ...ct UTIL The SELECT UTILITY OPTION screen will appear S E L E C T U T I L I T Y O P T I O N T E S T S S E T U P O U T P U T R E A D 3 From the SELECT UTILITY OPTION screen select TESTS The SELECT TEST...

Page 136: ...Dark 9911 10002 Delta 10595 25687 Lambda 405 Gain 2 53 Resets 2 Channel Ref 1 Air 20072 35592 Dark 9914 9960 Delta 10158 25632 Lambda 490 Gain 1 72 Resets 2 Channel Ref 1 Air 20280 35642 Dark 9919 994...

Page 137: ...the part numbers and versions of software currently loaded onto your reader If there are any errors during the power on checksum test they will be displayed 1 Press UTIL at the Main Menu At the SELECT...

Page 138: ...t of alignment If the reader fails the alignment test review the following possible problems and solutions Ensure that the Test Plate is correctly seated in the microplate carrier with A1 in the upper...

Page 139: ...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 r...

Page 140: ...1 1 040 1 050 1 040 0 881 0 783 H6 1 701 1 578 1 577 1 560 1 575 1 560 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...

Page 141: ...nu Select CALPLATE to access the CALIBRATION FILTER menu 3 Using the insert packaged with the Test Plate select the filter wavelength E D I T S E T U P I N F O R M A T I O N R S 2 3 2 C A L P L A T E...

Page 142: ...nce Plate Test ensure that the reader is not running in Rapid Mode To check this select UTIL READ and then cycle through the prompts until READ IN RAPID MODE is displayed Choose NO for an accurate res...

Page 143: ...cs Universal Plate Test 2 Select the Universal Plate ID and then click Run Test Test Failures If any of the test parameters report as FAIL confirm that the standard values on the test plate data sheet...

Page 144: ...have proof of linearity at wavelengths lower than those attainable with the Absorbance Test Plate This test is optional since the ELx800 has good front end linearity throughout the specified wavelengt...

Page 145: ...flask Preparation of Stock Solution 1 Weigh out 0 092 gram of FD C No 5 yellow dye powder into a weigh boat 2 Rinse the contents into a 1 liter volumetric flask 3 Add 0 5 ml of Tween 20 or 5 ml of Bio...

Page 146: ...into a 100 ml volumetric flask 2 Make up to 100 ml with DI water cap and shake well This should create a solution with an absorbance of about 2 000 when using 200 l in a flat bottom microwell The OD v...

Page 147: ...Column 1 of the plate 3 Pipette 200 l of the diluted solution into Column 2 of the plate 4 Read the microplate five times at 405 nm using the Normal Read Mode Chapter 3 single wavelength no blanking 5...

Page 148: ...viation of 0 0026 The mean 1 951 multiplied by 0 5 1 951 0 005 0 0098 which when added to the 0 005 0 0098 0 005 0 0148 OD which is the allowable deviation Since the standard deviation is less than th...

Page 149: ...ions 1 Set up a rack containing 10 tubes numbered consecutively 2 If you have not already done so prepare concentrated stock Test Solution A or B Refer to Table 3 when executing steps 3 and 4 Table 3...

Page 150: ...gnment Test 2 Print the raw data or export it to an Excel spreadsheet using KCjunior or KC4 If you are exporting the data to an Excel spreadsheet perform the calculations described below and on the fo...

Page 151: ...repeatability specification for a 96 well format of 0 5 0 005 OD from 0 000 to 2 000 OD 405 nm 3 The standard deviation for each set of readings should be less than the allowed deviation For example...

Page 152: ...r the H1 well to the same well in the A12 position The difference in the values for any two corresponding wells should be within the accuracy specification for the instrument For example If the mean o...

Page 153: ...filter installed in the reader New 96 well flat bottom microplates Corning Costar 3590 is recommended Calibrated hand pipette s Beakers and graduated cylinder Precision balance with readability to 0...

Page 154: ...oughly This is the 10x Concentrate PBS Solution Solution B Sigma PBS Required Materials Deionized water Tween 20 polyoxyethylene 20 sorbitan monolaurate Sigma P 3563 packets D NADH Powder D Nicotinami...

Page 155: ...ion as the diluent 3 Carefully prepare a 50 Test Solution by diluting 10 ml of the 100 Test Solution If using the Sigma PBS Solution use 10 ml as the diluent If using the 10x Concentrate PBS Solution...

Page 156: ...d deviation for well A1 is less than 0 017 the well meets the test criteria Linearity Test 1 For each of the three dye concentrations calculate the mean absorbance for the wells containing that soluti...

Page 157: ...aintenance and Decontamination This chapter contains procedures for maintaining and decontaminating the ELx800 Maintenance 130 Routine Cleaning Procedure 130 Replacing the Bulb 132 Decontamination Pro...

Page 158: ...id 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...

Page 159: ...e instrument from the power supply 2 Moisten a clean cotton cloth with water or with water and the mild detergent Do not soak the cloth 3 Wipe the plate carrier and all exposed surfaces of the instrum...

Page 160: ...s back and remove the screws holding on the top cover see Figure 3 in Chapter 2 3 Turn the instrument over again and lift the top cover off 4 Use a Phillips head screwdriver to remove the optics arm c...

Page 161: ...e Bulb 133 ELx800 Operator s Manual Optics arm cover Optics arm 1 aperture Lamp retention screws Lamp retention screws Bulb retainer Bulb assembly Optics arm Filter wheel cover Figure 20 Lamp assembly...

Page 162: ...turn the instrument off and on again to return the carrier to the home position Warning The alignment procedure requires you to observe the light path while the bulb is turned on To prevent possible...

Page 163: ...ters 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 procedure...

Page 164: ...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 cleanin...

Page 165: ...h a clean dry cloth 5 Wipe the plate carrier and all exposed surfaces of the instrument 6 Wait 20 minutes Moisten a cloth with deionized or distilled water and wipe all surfaces of the instrument that...

Page 166: ...138 Chapter 5 Maintenance and Decontamination Bio Tek Instruments Inc...

Page 167: ...s appendix describes potential error codes that may be displayed on the ELx800 and suggests possible solutions for these problems Overview 140 System Test Description 140 Glossary of Terms 140 Error C...

Page 168: ...powered on The reader will chirp repeatedly if the power on System Test results do not meet the internally coded Failure Mode Effects Analysis FMEA criteria established by Bio Tek A system test should...

Page 169: ...General Errors on the next few pages for more information Motor or Optical Sensor Channel 0 Carrier X Axis 0 Reference 1 Carrier Y Axis 1 Measurement A indicates a more serious error In this case tur...

Page 170: ...m KC4 User aborted from another serial interface 0200 X axis carrier in out motor did not find the opto home sensor A motor was unable to move to its home position as registered by feedback from an op...

Page 171: ...02 Filter wheel did not find opto home sensor Probable Cause Failed lamp 0300 0301 Light beam not found During the X axis movement the light beam saturation transition max light to no light was never...

Page 172: ...e same position when moved a known number of steps from the home position and back Probable Causes The belt has slipped because of incorrect tension a loose motor pulley or a loose belt clamp Defectiv...

Page 173: ...etrating the measurement chamber Ensure that the bottom and top shrouds as well as the filter wheel cover are correctly installed A faulty motherboard PN 7330400 7330410 or 7330415 or daughter board P...

Page 174: ...ad time Air Blank out of range 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 Lamp is u...

Page 175: ...otocol Probable Causes Assay wavelength does not match the filter table Run system test Verify that wavelengths in the filter wheel match settings in the filter table see Chapter 2 0F00 0F05 Filter si...

Page 176: ...l data exists for the read location Probable Cause Autocalibration was not performed following assay definition download 1300 Motor not homed successfully If the motor is not correctly homed the instr...

Page 177: ...Tek s KCjunior or KC4 PC Software This Appendix provides instructions for programming the computer to control the reader Overview 150 Controlling the Reader With KCjunior 151 Problems 152 Getting Star...

Page 178: ...ware provides the user even more power and flexibility For example complex assays using features not available through the front panel may be defined and run This Appendix provides instructions for co...

Page 179: ...M2 the serial port used for the RS 232 cable connection Baud Rate 2400 4800 or 9600 must match the baud rate on the reader Data Bits 8 Parity None Stop Bits 2 EOT Character Keep the default number 6 C...

Page 180: ...he Read Plate Dialog will appear 2 If desired enter a Results ID and a Plate Description and then click Read Plate The Protocol Definition dialog will appear 3 Select a Read Method Type of Endpoint Ki...

Page 181: ...nd subsequent Setup button to define the following communications parameters Port COM1 2 3 or 4 the serial port used for the RS 232 cable connection Baud Rate 2400 4800 or 9600 must match the baud rat...

Page 182: ...read a plate using KC4 1 Select Data New Plate 2 If prompted to select a protocol select Empty Protocol and click OK If not prompted select Protocol New or use KC4 s Protocol Wizard to step through pr...

Page 183: ...late data received from a plate with 384 well geometry To capture and manipulate the raw data using 384 well geometry you must use KCjunior or KC4 The Optical Density values will be downloaded via the...

Page 184: ...in 60 72 and 96 well Terasaki plate formats 1 Start KCjunior 2 At KCjunior s main menu choose Setup Reader 1 Figure 21 KCjunior main menu Setup Reader 1 dialog 3 The Reader Setup 1 dialog will appear...

Page 185: ...lick Read Plate to enter the Results ID and Plate Description Figure 23 Read Plate dialog 5 Choose New Protocol to define the protocol Figure 24 Protocol Definition General Information dialog ELx800 O...

Page 186: ...ing 384 Well Geometry 6 Define the Read Method parameters Set the Plate Geometry to 8 x 12 Figure 25 Protocol Definition Read Method dialog 7 Click OK to read the plate and display the results Bio Tek...

Page 187: ...96 well Terasaki plate formats 1 Start KC4 2 At KC4 s main menu choose System Readers Figure 26 KC4 main menu System Readers 3 The Reader Selection dialog will appear Select the ELx800 reader and defi...

Page 188: ...ate Figure 28 Data New Plate dialog 5 If prompted to select a protocol select Empty Protocol and click OK If not prompted select Protocol New or use KC4 s Protocol Wizard to step through protocol crea...

Page 189: ...rameters as necessary Click the Help button for assistance 11 When complete click OK 12 Select Data Read Plate The Plate Reading dialog will appear 13 Enter any comments place the plate on the carrier...

Page 190: ...162 Appendix B Using 384 Well Geometry Bio Tek Instruments Inc...

Page 191: ...reports that can be generated and or printed from the ELx800 Refer to Printing Reports in Chapter 3 for details on how to print these reports In addition an Assay List Assay Definition Map and Result...

Page 192: ...164 Appendix C Report Format Figure 31 Samples with calls on Matrix Report Bio Tek Instruments Inc...

Page 193: ...165 Figure 32 Curve Fit Report ELx800 Operator s Manual...

Page 194: ...166 Appendix C Report Format Figure 33 Samples with calls on Column Report Bio Tek Instruments Inc...

Page 195: ...167 Figure 34 Column Report without samples ELx800 Operator s Manual...

Page 196: ...168 Appendix C Report Format Figure 35 Panel Report Bio Tek Instruments Inc...

Page 197: ...169 Figure 36 Assay Detail Report Sheet 1 of 2 ELx800 Operator s Manual...

Page 198: ...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 3...

Page 199: ...ates x 48 well plates x 96 well plates x x Edit Assay Name x Absorbance Values x x Formulas x Curve fits x Assays available 55 10 Results stored 8 1 Blanks and samples only in Map x Blanks standards c...

Page 200: ...172 Appendix D Comparison of the ELx800 and the EL800 Bio Tek Instruments Inc 172...

Page 201: ...ructions are provided so that users can see how it is possible to translate the kit wording into an ELx800 assay program For clarity only the user menu choices from the reader screens are shown Refer...

Page 202: ...d 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 di...

Page 203: ...ng 1 8 ml of diluted stabilized human serum Ready to use ANA Conjugate 1 bottle containing 12 ml of peroxidase labeled antibody to human IgG Colored red ready to use TMB Substrate 1 bottle containing...

Page 204: ...ay 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...

Page 205: ...TO Auto mapping is normally preferred because it fills in the well IDs logically and automatically after determining which direction to map and how many wells to fill DOWN Maps the wells down the colu...

Page 206: ...indicate the average of the Positive Control replicates 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...

Page 207: ...n 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 precoat...

Page 208: ...ds 5 bottles each containing 1 2 ml of diluted human serum with the following concentrations of anticardiolipin autoantibody 100 50 25 12 5 6 25 GPL U ml Ready to use The standard set is calibrated ag...

Page 209: ...ol and Results Quality Control For an assay to be valid all the following criteria must be met Standards and the positive and negative controls must be included in each run The values obtained for all...

Page 210: ...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 c...

Page 211: ...ls are higher still at 17 0 IgM and 10 6 IgG Expected Values The normal range was determined on serum from 102 normal adult blood donors The ranges below are provided as a guide only ELISA assays are...

Page 212: ...llow steps below STEP COMMENTS To program the reading method press METHOD WAVELENGTH Dual or Single Single MEASURE Wavelength s to use 450 PLATE TYPE 96 To program the plate map press MAP AUTO DOWN DO...

Page 213: ...ue 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 Curve 4P...

Page 214: ...186 Appendix E Instructions for Programming a New Assay Bio Tek Instruments Inc...

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