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Appendix B

B-23

TEMPERATURE POLLING

The

Get Temperature

(‘]‘) command is defined as follows:

Host Response

']'

The Reader response protocol (hex format):

temperature low byte

0x00-0xFF

temperature high byte

0x00-0xFF

The

Get Temperature

command then returns the standard status response string.

Temperature is returned scaled up by 10, i.e. 370 indicates a temperature of 37.0

degrees Celsius. If the instrument does not have incubation, or an incubation error

has been detected, 0x00, 0x00 (hex, not ASCII) will be returned as temperature data.

An 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 should be able to 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 <^Z> checksum combination has been sent.

KINETIC INTERVALS

Summary of Contents for PowerWave

Page 1: ...PowerWaveTM Microplate Scanning Spectrophotometer Operator s Manual ...

Page 2: ... ...

Page 3: ...1 Unpacking and repackaging the reader Carefully open the top of the box and remove the dust cover The box will include a power cord an Operator s Manual and optionally a Service Manual Remove the end cap from the top of the reader Lift the reader out of the box and place it on a level surface Remove the reader from the plastic bag Place all shipping material back into the shipping box for reuse i...

Page 4: ......

Page 6: ...ii ...

Page 7: ...retrieval system or translated into any language without the written permission of Bio Tek Instruments Incorporated Trademarks Bio Tek is a registered trademark and PowerWave 200 PowerWave 340 Extensions RoboReader KCJr KCII KC3 KC4 ELx 800 and ELx 808 are trademarks of Bio Tek Instruments Inc Windows is a trademark of Microsoft Inc Restrictions and Liabilities Information in this document is subj...

Page 8: ...eferences to monochronometer to monochromator B1 8 97 Inserted a Manual Update correcting the calibration plate Part Number C 8 97 Corrected Cal Plate PN in Chapters 1 and 4 to 7260522 Corrected Error Codes in Appendix C Added accuracy and repeatability data to Optical Specifications section Added Computer Control commands to Appendix B Printed in the United States of America Bio Tek Printed Mater...

Page 9: ...Avoid spilling liquids on the reader fluid seepage into internal components creates a potential shock hazard Do not operate the instrument if internal components are exposed to fluid Warning Software The microplate reader operator must follow the manufacturer s assay package insert when modifying software parameters and establishing result calculation methods using the reader s on board software W...

Page 10: ...y be adversely affected if 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 contact may damage the instrument surfaces Be certain to rinse and thoroughly wipe all surfaces Caution Power Supply Only use the correct line voltage when operating the Automated M...

Page 11: ...rgy 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 ...

Page 12: ... Susceptibility Verification of compliance was conducted to the limits and methods of EN 50082 1 1992 IEC 1000 4 2 1995 IEC 1000 4 3 1995 and IEC 1000 4 4 1995 Safety This device has been type tested by an independent laboratory and found to meet the requirements of North America Canadian Standards Association CAN CSA C22 2 No 1010 1 1992 Safety Requirements for Electrical Equipment for Measuremen...

Page 13: ... manual carefully before operating this instrument Alternating current Courant alternatif Dreiphasen Wechselstrom Corriente Atterna Corrente alternata Earth ground terminal Borne de terre Erde Bettriebserde Borne de Tierra Terra di funzionamento Protective conductor terminal Borne de terre de protection Schutzleiteranscluss Borne de Tierra de Protección Terra di protezione On Supply Marche aliment...

Page 14: ...cuments d accompanement Achtung siehe Begleitpapiere Atención vease los documentos incluidos Attenzione consultare la doc annessa Caution risk of electric shock Attention risque de choc electrique Gefährliche elektrische Spannung Atención riesgo de sacudida eléctrica Alta tensione in questo documento Alta tensione non significa tensione pericolosa come definito in IEC 417 ...

Page 15: ...rve fitting and statistical software program is preloaded on every PowerWave 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 KC Jr or KC4 for Windows The software provides An easy to use menu driven interface Endpoint curvilinear regressional and statistical calcul...

Page 16: ... the beginning of this paragraph Major topic headings start a new page such as About This Manual above to give you a visual and style clue that a new major subject is being introduced One or more subheadings may appear below each major heading Registration Card Once the Microplate Reader has been set up and is running successfully please take a moment to fill out and mail the postage paid Warranty...

Page 17: ...thout cost to the Purchaser any Product which in Bio Tek s sole judgment is defective by reason of defects in the materials or workmanship This Warranty is VOID if the Product has been damaged by accident or misuse or has been damaged by abuse or negligence in the operation or maintenance of the Product including without limitation unsafe operation operation by untrained personnel and failure to p...

Page 18: ...xiv ...

Page 19: ...ng and Repackaging the Instrument 2 1 After Unpacking Verify Performance 2 3 Before Repackaging the Instrument 2 3 Adjusting the Reader s Wavelength Table Setting 2 4 Printing and Data Communications 2 6 Installing a Printer 2 6 Parallel Port Pin Definition 2 7 Setting Up the Serial Port Communications with Other Devices 2 8 Attaching the Serial Cable 2 8 Serial Port Pinout Description 2 9 Setting...

Page 20: ... Main Menu Screen 3 3 Define 3 4 Select Assay 3 4 Assay Name 3 5 Define Method Map Formula and Curve 3 6 Method 3 6 Read Type 3 6 Delay in First Read 3 7 Incubation Temperature 3 8 Single or Dual Wavelength 3 9 Wavelength Selection 3 10 Number of Kinetic Reads Kinetic Duration 3 10 Kinetic Interval 3 11 Kinetic Number of Reads 3 12 Kinetic Duration 3 12 Shake Mode Selection 3 12 Shake Time 3 13 Sh...

Page 21: ...lue 3 24 Number of Blanks 3 25 Blank Location 3 25 Number of Standards 3 26 Number of Standard Replicates 3 26 Average Standards 3 27 Standard Concentration 3 27 Reuse of Standard Curves 3 28 Number of Controls 3 30 Type of Control 3 31 Number of Control Replicates 3 31 Location of Controls 3 32 Number of Samples 3 32 Number of Sample Replicates 3 33 Sample Location 3 33 Panel 3 35 Formula Definit...

Page 22: ...7 Transformation Scope Variable 3 48 Defining the Curve Fit 3 52 Curve Fit 3 52 Edit Standard Outliers 3 54 Axis Selection 3 55 Extrapolation of Unknowns 3 55 Reading a Microplate 3 57 Select Assay 3 57 Run Time Prompts 3 58 Enter Number of Samples 3 59 Enter Plate ID 3 59 Enter Sample ID 3 60 Prompts for Well Location 3 60 Beginning the Plate Read 3 61 Printing Reports and Assay Lists 3 62 Result...

Page 23: ...5 System Self Test 4 6 Checksum 4 8 Calibration Validation 4 9 Calibration Plate Test 4 9 Requirements 4 9 Entering Calibration Plate Specifications 4 10 Running the Calibration Plate Test 4 12 Calibration Plate Analysis 4 13 Linearity Test 4 15 Equipment Required 4 15 Procedure 4 16 Verification of Alignment 4 17 Appendix A Decontamination A 1 Appendix B Computer Control B 1 Appendix C Error Code...

Page 24: ...xx ...

Page 25: ...capability of spectral scanning using a monochromator This allows the user to perform endpoint and kinetic analysis in 96 well microplates using any wavelength between 200 and 800 nm for PowerWave 200 and 340 to 800 nm for PowerWave 340 thereby achieving the maximum absorbance for any sample The reader features superior optical specifications with an extended dynamic range of up to 4 000 absorbanc...

Page 26: ...s according to instructions and specific recommendations included in the package insert for the test to be conducted Failure to conduct Quality Control checks could result in erroneous test data Regulatory Agency Approvals The instrument has been tested approved and externally labeled to show compliance to the following test standards ETL CAN CSA C22 2 No 1010 1 92 EN 61010 1 CE EN50081 1 Emission...

Page 27: ...arallel port 25 pin female connector Internal bar code option Four zone incubation chamber option Robotics interface option Software Features _______________________________________________ Easy to use menu driven interface Endpoint Kinetic and Scanning calculations Curve fitting with 4 parameter cubic quadratic linear 2 P cubic spline and point to point methods Transformation and formula calculat...

Page 28: ...Wave package includes Microplate Reader Power Cord Operator s Manual and Warranty Registration Card Dust cover PN 7342066 Parallel cable PN 71072 Optional Accessories Service Manual for all instrument models PN 7261001 Replacement lamp assembly PN 49789 Serial cable PN 75053 Calibration Test Plate PN 7260522 ...

Page 29: ...rbance Measurement Range 0 4 000 Abs 200 340 900 nm Accuracy 1 010 Abs from 0 to 2 000 Abs 3 010 Abs from 2 000 to 3 000 Abs measured with neutral density filters calibration test plate 405 450 490 550 630 690 nm Linearity 1 from 0 to 2 000 Abs 3 from 2 000 Abs to 3 000 Abs measured 405 nm Repeatability STD 1 005 Abs from 0 to 2 000 Abs 3 from 2 000 Abs to 3 000 Abs measured 405 nm Fast mode of re...

Page 30: ...e 1 100 VAC 90 110 VAC 50 60 Hz Range 2 120VAC 103 127 VAC 50 60 Hz Range 3 230 VAC 207 253 VAC 50 60 Hz Range 4 240VAC 216 264 VAC 50 60 Hz Incubation optional Temperature Control 4 over ambient to 50 C Temperature Variation 0 5 C 37 C with the plate sealed Optional internal Bar Code reader If enabled the internal bar code reader recognizes a number of common bar code types during the plate read ...

Page 31: ...R allows the reader to function with an autoloading robot Using computer control commands from a host PC the reader s functions can be controlled in conjunction with the robotic system The Robot interface model can be configured with all options available for the standard PowerWave ...

Page 32: ...802 655 4044 European 32 3 829 0455 Emergency Assistance is available Monday through Friday 6 00 PM to 9 00 PM EST Fax 802 655 3399 E mail labtac biotek com INTERNET http www biotek com You may write a letter with your comments and send it to Bio Tek Instruments Technical Assistance Center Highland Park Box 998 Winooski VT USA 05404 0998 Lionheart Technologies Europe N V Sneeuwbeslaan 33 Wilrijk B...

Page 33: ...can reduce the linear performance range and affect the instrument s readings Dust Optical density readings may be affected by extraneous particles such as dust in the microplate wells A clean work area is necessary to ensure accurate readings Unpacking and Repackaging the Instrument If the shipping box has been damaged inspect the instrument for visible dents and scratches as you unpack it If the ...

Page 34: ... and mark the RMA number on the outside of the shipping container Figure 2 1 illustrates how to unpack the reader Figure 2 1 Unpacking and packing the reader Carefully open the top of the box and remove the dust cover The box will include a power cord an Operator s Manual and optionally a Service Manual Remove the end caps from the reader Lift the reader out of the box and place it on a level surf...

Page 35: ...trument Decontaminate the reader prior to shipping Refer to the Decontamination procedure in Appendix A Once the reader is clean pack it in its original shipping box using original packing materials This shipping system was designed to be used no more than five times If the container is damaged and or has been used more than five times contact Technical Services for a new set of shipping materials...

Page 36: ... Option Screen From the Select Utility Option menu press the soft key beneath the menu option SETUP The Edit Setup Information screen is displayed Figure 2 4 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 L A M B D A m o r e Figure 2 4 The Edit Setup Screen From this menu press the soft key under the LAMBDA menu option The wavelength will be displayed Figure 2 5 To advance to the next wav...

Page 37: ...avelength Entry screen When the last wavelength has been entered the software exits the routine and displays the following screen Figure 2 6 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 L A M B D A M O R E Figure 2 6 The Edit Setup screen Press the Main Menu key to return to the Main Menu ...

Page 38: ...graphics compatibility The reader s parallel port requires a 25 pin D sub connector Figure 2 7 Serial and Parallel connectors The parallel port s pinout definition is described in Table 2 1 A printer cable is supplied with the reader Should the cable that came with the reader become lost or damaged Bio Tek offers replacement printer cables Contact your authorized Bio Tek dealer for information on ...

Page 39: ...are tightened and power up the reader and printer iii IMPORTANT To avoid system instability connect the printer to the reader before powering up the reader Turn on the PowerWave first then the printer Parallel Port Pin Definition Table 2 1 illustrates the pin definitions for the reader s 25 pin socket female D sub Parallel connector Parallel Port Pinout Pin Signal Pin Signal 1 PSTROBE 14 NC 2 D0 1...

Page 40: ...RS232 protocols Appendix B contains information on required protocols for computer control of the reader Attaching the Serial Cable Power down the computer and the reader Connect the appropriate serial cable to both machines The serial port on the reader is a DTE configuration with a 25 pin pin male D sub connector The connector s pinout is illustrated in Table 2 2 Power up the reader and the comp...

Page 41: ...ations Parameters Before serial communications are initiated between the PowerWave 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 9600 Baud 8 Data Bits 2 Stop Bits and No Parity The user may change the Baud rate on the reader from the recommended default setting of 9600 ...

Page 42: ... the UTILITY OPTIONS screen press the soft key beneath the menu option SETUP Figure 2 9 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 Figure 2 9 Selecting Setup At the EDIT SETUP INFORMATION screen press the soft key beneath the MORE menu option to advance to the EDIT SETUP RS232 option screen Figure 2 10 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 L A M...

Page 43: ...2 0 0 2 4 0 0 9 6 0 0 V I E W Figure 2 11 Selecting the Baud Rate 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 To view the reader s other communication settings Figure 2 12 press the soft key beneath the VIEW menu option R S 2 3 2 S E T T I N G S N O P A R I T Y 2 S T O P B I T S 8 D A T A B I T S Figu...

Page 44: ... will slowly drop over time until the reader s run time self check detects a low signal and warns the user with a displayed message The bulb should be replaced at this time using a replacement bulb part numbered PN 49789 Refer to Figure 2 13 for a detailed view of access to the lamp assembly Figure 2 13 Location of the lamp assembly Bulb Housing Bulb Socket Bottom Plate Mounting Hardware ...

Page 45: ...fore proceeding Remove the bottom plate mounting hardware 6 Remove the bulb socket mounting hardware 2 and carefully remove the bulb housing Remove the old bulb from the socket Using lens paper or gloves replace the bulb and mount it back into the bulb housing There should be no fingerprints on the bulb Replace the bottom using the mounting hardware ...

Page 46: ...tting 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 Figure 2 14 illustrates the power input module Figure 2 14 Selecting voltage input Unplug the reader and remove the power cord Use a small f...

Page 47: ...itch board 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 Fus...

Page 48: ...o replace a bad 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 t...

Page 49: ...e Counts TESTS System which checks reader optics CHKSUM which checks version of software installed CALPLATE which runs Calibration Plate Test See page 4 5 for more information To set these user configurable options At the Main Menu screen press the soft key beneath UTIL to access the Utility Options menu 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 Figure 2 15 Sele...

Page 50: ...NTER to return to the EDIT SETUP INFORMATION screen To edit the TIME press the soft key positioned beneath the TIME menu option At the Time entry screen Figure 2 18 use the NUMERIC keys to enter the correct time The cursor is positioned at the first editable field T I M E 0 3 1 1 P M 1 2 H R 1 2 H O U R 2 4 H O U R A M P M Figure 2 18 Entering the Time Selecting the Format Select a 12 or 24 hour f...

Page 51: ... advance to the REPORT TYPE menu screen Figure 2 20 R E P O R T T Y P E M A T R I X C O L U M N M A T R I X B O T H Figure 2 20 Selecting Report Type Any previously defined 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 Figure 2 21 S A M P L E S I N C...

Page 52: ...ompt preferences Figure 2 23 Select YES to prompt for identifications and sample counts before a microplate is read Press the soft key beneath YES or NO to select Plate ID prompts for microplate identification Sample ID will prompt for an identification for each sample Sample Count will ask for the number of samples on each plate Pressing ENTER after each selection advances the display When select...

Page 53: ... 2 X 24 character LCD display allowing the user to access the reader s program menus to view test results The reader s bi directional serial port allows computer control of the instrument and provides the means for downloading additional assay definition files to the instrument The Keyboard Figure 3 1 shows the instrument s keyboard layout Figure 3 1 Keyboard ...

Page 54: ...ture sensitive testing is being completed as it may introduce dust into the internal workings of the instrument The Startup Screen The PowerWave performs a self test when powered on displaying the Startup screen until initialization is complete During this period no keys are active Should the instrument fail to pass the self test a beep will sound and an error code will display Refer to Appendix C...

Page 55: ...ponds to a displayed menu option to activate that option READ option or press the key labeled READ on the keyboard for plate reading prompts The user will be asked to select an assay from a list of choices DEFINE option Allows the creation of a reading and data reduction protocol The user will be prompted to select an assay from an assay definition list REPORT option Allows printing of results and...

Page 56: ...nge depends on the number of assays programmed in the reader s memory The reader has 55 open assays available The assay s name and number are displayed on the screen Figure 3 3 S E L E C T A S S A Y N U M B E R 0 1 N A M E H B S A G 1 Figure 3 3 Assay Selection screen Press ENTER to advance to the EDIT ASSAY NAME screen You may change the default assay number to a more descriptive one see assay na...

Page 57: ... change the Assay name The cursor is positioned at the first editable field Use the OPTION key to sequentially advance the character positioned above the cursor The characters will cycle through the alphabet A Z with a space following Z Use the LEFT RIGHT ARROW keys to move the cursor to the previous or next editable field The cursor will wrap around the edit field Use SOFT KEYS 1 2 3 4 when using...

Page 58: ...arameters Kinetic analysis iii NOTE Some screens shown here may not appear on some reader models READ TYPE This option allows the user to enter which read type Endpoint Kinetic or Scan Endpoint The plate is read once The unit of measure is Optical Density Kinetic The plate is read several times Three different types of data are available a For a rate determination the data calculated is the rate o...

Page 59: ...canned divide the scanned OD result by the total number of scans The following keys are active during this screen Figure 3 6 R E A D T Y P E K I N E T I C E N D P O I N T K I N E T I C S C A N Figure 3 6 Read Type screen Use SOFT KEY 1 to select Endpoint read mode Use SOFT KEY 2 to select Kinetic read mode DELAY IN FIRST READ ENTRY Selecting the Delay in First Read option allows the user to enter ...

Page 60: ...M P E R A T U R E Figure 3 8 Incubation Temperature entry Use SOFT KEY 1 or 2 to select ambient incubation Use SOFT KEY 3 or 4 to select an adjustable temperature Use the LEFT and RIGHT ARROW keys to move the cursor between the two digits on the input temperature Use NUMERIC keys to enter the incubation temperature The range is 25 to 50 C Incubation performance is specified from four degrees over ...

Page 61: ... the difference between the two readings iii NOTE Dual wavelength readings significantly reduce optical interference caused by scratches or fingerprints which absorb light equally at both wavelengths Choose a REF filter that is far from the MEAS filter in the spectrum for best results Any previously defined wavelength appears on the top line of the display and the options on the second W A V E L E...

Page 62: ...option displayed above the soft key The display updates to reflect the selection Use the LEFT or RIGHT ARROW keys to move the cursor between the measurement and reference filter selections NUMBER OF KINETIC READS KINETIC DURATION SELECTION This menu allows the user to either select the total number of kinetic reads or the length of time the assay will run kinetic duration Figure 3 11 Any previousl...

Page 63: ... minutes and seconds between each kinetic read K I N E T I C I N T E R V A L 0 1 2 3 5 6 Figure 3 12 Kinetic Interval entry screen 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 or equal to 2 Use the LEFT and RIGHT ARROW keys to move to the next or previous...

Page 64: ...4 to enter in hours minutes and seconds the duration of the kinetic reaction K I N E T I C D U R A T I O N 1 1 2 3 4 5 Figure 3 14 Kinetic Duration entry screen Use the NUMERIC keys to enter the time duration in hours minutes and seconds The maximum duration time is 80 hours Use the LEFT and RIGHT ARROW keys to move between entry fields SHAKE MODE SELECTION Use this screen Figure 3 15 to enter the...

Page 65: ...keys to enter the shake interval Valid ranges are 0 1 hours 0 59 minutes and 0 59 seconds Press SOFT KEY 1 to select CONTINUOUS SHAKE The shake time will be set automatically according to the kinetic interval Use the LEFT and RIGHT ARROW keys to move the cursor between hours minutes and seconds SHAKE SPEED Use this screen Figure 3 17 to select the shake speed The shake movement is a repeated 021 i...

Page 66: ... R R A T E R S Q R O N S E T Figure 3 18 Kinetic Data Analysis selection screen Use SOFT KEY 1 to select the kinetic rate calculation This method will apply a linear fit to calculate the maximum slope in mOD min based on the number of kinetic points specified Use 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 lin...

Page 67: ...o MAX where max is the total number of reads Use SOFT KEY 1 or 2 to select All points iii Note If the number of kinetic reads is changed during assay definition be certain also to adjust the kinetic points The number of kinetic points must be equal to or less than the total kinetic reads ONSET OD SELECTION Use this screen Figure 3 20 to enter the onset OD value E N T E R O N S E T O D 1 2 3 4 Figu...

Page 68: ...e 31 scan positions are fixed in the software The user must determine the optional 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 iii Note If too many scans are chosen the reader may be reading the sides of the well The reader will rea...

Page 69: ...p Selection Blanking constant Number of Blanks Location of Blanks Number of Standards Number of Standard Replicates Averaging of Standards Concentration and Location of Standards Number of Controls Control Type Definition Number of Control Replicates Control Location Number of Samples Number of Sample Replicates Sample Location 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 Figure 3 22 Sele...

Page 70: ...or Automatic Map Generation Automatic Plate Map Generation refers to the ability of the software to automatically generate a Plate Map after the number of blanks controls standards and samples have been identified Manual Plate Map Generation refers to the ability of the user to identify where each blank control standard or sample will be located Press SOFT KEY 1 for sample plate map generation The...

Page 71: ...ne see Figure 3 25A M A P P I N G D I R E C T I O N D O W N D O W N A C R O S S Figure 3 24 Mapping Direction Entry screen Press SOFT KEY 1 to map DOWN the column Press SOFT KEY 2 to map ACROSS the row REPLICATION DIRECTION This option Figure 3 25 allows the user to specify how replicates are mapped on the plate Any previously defined Replication Direction appears on the top line of the display an...

Page 72: ...of the map listing 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 Examples of mapping directions are shown in Figure 3 25A ...

Page 73: ...B NC NC SMP SMP SMP SMP SMP SMP C D E F G H The map above shows Map Direction ACROSS Rep Direction ACROSS 1 2 3 4 5 6 7 8 9 10 11 12 A STD1 STD1 B STD2 STD2 C STD3 STD3 D STD4 STD4 E STD5 STD5 F PC PC G NC NC H SMP SMP The map above shows Map Direction DOWN Rep Direction ACROSS 1 2 3 4 5 6 7 8 9 10 11 12 A STD1 STD2 STD3 STD4 STD5 PC NC SMP SMP B STD1 STD2 STD3 STD4 STD5 PC NC SMP SMP C D E F G H ...

Page 74: ...the cursor to the previous or next editable field The cursor will wrap around the edit field Use the NUMERIC 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 character BLANK MAP This option Figure 3 27 allows the user to select which blanking method to apply t...

Page 75: ...al blanking value on air as a blank value FULL enables a single blank well or an average of blank wells up to 48 to be subtracted from the whole plate CONST Constant allows entry of a user specified absorbance value This value wil be subtracted from each well on the plate ROW enables a single blank well or an average of blank wells to be selected for each row The maximum number of blanks is 48 Man...

Page 76: ...al mapping to set up the appropriate map by placing the standards controls and samples in only the A C E and G rows CONSTANT BLANK VALUE This entry screen Figure 3 28 only appears when a Constant Blank map is selected The user enters a value that is subtracted from each well on the plate E N T E R B L A N K I N G C O N S T A N T 1 2 0 Figure 3 28 Constant Blank Value screen Use the NUMERIC keys to...

Page 77: ... 48 Use the CLEAR key to clear the Number of Blanks value from the display BLANK LOCATION The Blank Location screen Figure 3 30 allows the user to define where the blank well occurs on the microplate This screen only appears if Manual Map Generation has been selected Any previously defined value is displayed E N T E R T H E L O C A T I O N O F B L A N K 1 A 1 2 Figure 3 30 Defining the Location of...

Page 78: ...ected curve fit method The maximum number of standards is 12 The minimum is 4 for 4 P fit cubic cubic spline and logit log 3 for quadratic and 2 for linear and point to point NUMBER OF STANDARD REPLICATES This option Figure 3 32 allows the user to enter the number of replicates per standard in the assay Any pre defined value appears on the display E N T E R N U M B E R O F S T D R E P L I C A T E ...

Page 79: ...on Use SOFT KEY 2 to select NO do not average the replicates The top line of the display updates to reflect the selection STANDARD CONCENTRATION The Standard Concentration field Figure 3 34 allows the user to enter a concentration value for each standard Any previously defined value is displayed If automatic map generation is selected the location field is only editable for the first replicate of ...

Page 80: ...mitations of the Reuse of Standard Curves The reuse of standard curves can only be done in assay positions 51 through 55 Each of these positions can only store one standard curve Only 5 curves can be stored at any one time In order for a standard curve to be reused the assay name from the stored curve and the name of the assay being run must be identical Standard curves cannot be reused on panels ...

Page 81: ...ts have been generated the reader will prompt if this standard curve should be stored in memory The following display will appear Figure 3 36 S A V E S T A N D A R D C U R V E Y E S Y E S N O Figure 3 36 Save Standard Curve screen Choosing YES will store the curve to be used at a later time The next time this assay is used the instrument will prompt if there are standards on the plate Figure 3 37 ...

Page 82: ... 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 re used NUMBER OF CONTROLS The Number of Control screen Figure 3 38 allows the user to enter the number of controls that will be used in the assay Any previously defined value will appear on the display E N T E R N U M B E R O F C O N T R ...

Page 83: ...C C T L 3 C T L 4 M O R E Figure 3 39 Types of Control screen 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 CTL4 Use the CLEAR key to clear the Control Type from the display NUMBER OF CONTROL REPLICATES The Number of Control Replicates screen Figure 3 40 allows the user to enter ...

Page 84: ... is editable C O N T R O L 1 L O C A T I O N T Y P E P C R E P 1 A 0 2 Figure 3 41 Location of Controls screen Use the CLEAR key to clear the value on the display Use the NUMERIC ALPHA keys Use these keys to enter characters and numeric values for valid well locations depending on the plate geometry Valid locations are VALID WELL LOCATIONS For all prompts of a well location only the ALPHA keys are...

Page 85: ... value will be displayed E N T E R N U M B E R O F S A M P L E R E P L I C A T E S 2 Figure 3 43 Number of Sample Replicates screen 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 does not exceed the number of undefined wells remaining on the plate SAMPLE LOCATION Th...

Page 86: ...ample and its location on the plate Valid characters and numeric entries are based on the selected plate geometry The following are acceptable entries for well locations based on plate geometry 96 well plate range A01 A12 B01 B12 C01 C12 D01 D12 E01 E12 F01 F12 G01 G12 and H01 H12 ...

Page 87: ... will be used when the assay is selected for a Panel Panel assays cannot re use standard curves The type and number of controls blanks standards and replicates in the assays chosen for the Panel will be copied into the Panel definition If the user wishes to change any of the map or assay parameters they must be changed in the full assay first To use the Panel option press DEFINE and choose Assay N...

Page 88: ...er of assays comprising this panel N U M B E R O F A S S A Y S 2 Figure 3 46 Number of Assays screen Use the NUMERIC keys to update the display and the cursor to advance to the next field The limit is 8 assays The next screen Figure 3 47 when defining a panel prompts to select the mapping direction of the assays The available options are DOWN or ACROSS iii NOTE All original map directions for the ...

Page 89: ...hoose which assays will be used in the panel Figure 3 48 It is important to note that these assays must have been previously defined to be used in the panel definition and must all use the same wavelength S E L E C T A S S A Y N U M B E R 2 2 N A M E H B S A G 1 Figure 3 48 Selecting the Assay Number The OPTION key will increment the assay number by one The display will be updated as the keys are ...

Page 90: ...the Assay setups for reference before mapping them in the Panel For example if the user wishes to run Assays 1 8 and 22 within a Panel Assay 1 has a total of 12 wells defined for controls blanks and standards and in the Panel the mapping for Assay 1 begins in well A01 with the map direction DOWN The user wants to run 6 samples in Assay 1 Assay 1 now fills wells A01 through B03 The mapping for Assa...

Page 91: ...PowerWave Operator s Manual 3 39 Figure 3 49A Panel Report ...

Page 92: ... 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 Validation 0 1 Control Validation 0 4 Assay Validation 0 4 Transformations 0 1 Cut off Formulas 0 1 General Formulas 0 4 If a curve fit method is also specified here s the order in which the processing occurs Blank Validation Control Validation Assay...

Page 93: ...EY 2 to select Transformation formula Press SOFT KEY 3 to select Assay validation formula Press SOFT KEY 4 to select General Formula VALIDATION TYPE SELECTION Use this option to select which Validation Formula type Control Assay or Blank Validation formulas to enter for the assay Figure 3 51 S E L E C T V A L I D A T I O N T Y P E C O N T R O L A S S A Y B L A N K Figure 3 51 Validation Type selec...

Page 94: ...igure 3 52 Formula entry screen Use LEFT and RIGHT ARROW keys to move the cursor to the previous or next editable field Press SOFT KEY 1 to place the next item on the MATH list at the cursor position See the table that follows for the order of items on the MATH list MATH LIST Addition sign Subtraction sign Multiplication sign Division sign Percent Equal Greater then Greater than or equal to Less t...

Page 95: ...Press SOFT KEY 3 to place the next defined item on the plate map list i e STD NC PC BLK at the cursor position Press SOFT KEY 4 to place the next option on the FUNCTION list at the cursor position See the table that follows for the order of items on the FUNCTION list FUNCTION LIST LOG10 Log Base 10 ALOG10 Ante Log Base 10 AB Absolute Value PWR Power ALOG Anti Log LOG Log SQRT Square Root Functions...

Page 96: ...Validation Formula NC 0 100 Positive Control Validation An assay protocol states that the Positive Control wells must have an OD higher than 1 000 but less than 2 500 Two Control Validation Formulas can be entered PC 1 000 and PC 2 500 Or one formula can be used if the formula is 24 characters or less PC 1 000ANDPC 2 500 Assay Validation An assay protocol states that in order for an assay to be va...

Page 97: ...ve Similar to entering validation formulas cutoff formulas use the same math functions EXAMPLE A cutoff between negative and positive could be the average of the negative controls added to an absorbance of 0 050 This would be NC x 0 050 If a user wishes to define a cut off value that determines whether a well would be called POS or NEG or EQUIV then the user would define a Cutoff Formula For examp...

Page 98: ...N T E R G R A Y Z O N E 0 5 Figure 3 54 Grayzone Percentage screen Use the NUMERIC keys to enter the grayzone value The range is 1 to the number of defined replicates of a control POSITIVE OR NEGATIVE CALL FOR CUTOFF The positive or negative call for cutoff allows the user to define the calls above cutoff as positive or negative Figure 3 55 Calls below the cutoff will be the reverse of the above c...

Page 99: ...w corresponding to Formula Figure 3 56 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 Ï Figure 3 56 The Define screen This will bring the user to a screen asking to Select Formula Type At this screen Figure 3 57 select TRANS Then enter the formula using the Math Other Map and Function keys S E L E C T F O R M U L A T Y P E C U T O F F T R A N S V A L M O R E Ï Figure 3 57 Select Formula Typ...

Page 100: ...ples SMP or to all wells defined on the plate OD From the assay Define Menu press the arrow corresponding to Formula 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 Ï This will bring the user to a screen asking to Select Formula Type At this screen press MORE S E L E C T F O R M U L A T Y P E C U T O F F T R A N S V A L M O R E Ï The options displayed now include Trans Var Select the arrow t...

Page 101: ...tifiers CTRL PC NC STD etc defined will become available as MAP options when writing the transformation formula EXAMPLE 2 An assay map contains an NC and a CTRL1 both in duplicate The remainder of the map is filled with samples The assay data reduction requires that the mean of the NC be subtracted from all the SAMPLES on the plate but not from the CTRL1 On paper the formula reads SMP NC x To prog...

Page 102: ...an of CTL1 from all OD s on the plate Divide by the mean of the NC minus the mean of CTL1 then multiply by 100 On paper the formula reads OD NC x CTL1 x NC x CTL1 x 100 On the reader the formula NC x CTL1 x will be programed as the Transformation Variable since the transformation will apply to all standards controls and samples on the plate To do this At the SCOPE VARIABLE selection choose OD and ...

Page 103: ...00 This would divide all the wells by Std1 presumably the 0 standard and multiply by 100 Go to TVAR choose OD Select STD1 from MAP At transformation formula screen enter OD TVAR 100 GENERAL FORMULA A general formula is any desired data reduction that may accompany other reports This feature is not used in the current version of the reader ...

Page 104: ... 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 sim...

Page 105: ... 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 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 t...

Page 106: ...ect 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 EDIT STANDARD OUTLIERS This screen Figure 3 61 allows the user to select which method None or Manual will be used to edit Standard Outlier values Any previously defined edit method is displayed E D I T...

Page 107: ...xis identification option displayed above the soft key The top line of the display updates to reflect the selection EXTRAPOLATION OF UNKNOWNS This screen Figure 3 63 allows the user 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 Not available for all curve fit types E X T ...

Page 108: ...olated 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 ...

Page 109: ...efined 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 Figure 3 64 S E L E C T A S S A Y N U M B E R 6 5 N A M E H B S A ...

Page 110: ...ssays were created typical prompts might include The number of samples Standard concentrations Assay ID Fill pattern Blank method First well location Replicate count for each well type Wavelength mode Report preferences etc Refer to the Extensions Reader Protocol Manual for more information on user prompts that might be encountered Utility read options might include Enter number of samples Plate I...

Page 111: ...be unique iii NOTE Use caution when creating multiple Plate IDs The reader does not warn the user that he is about to exceed the maximum of 10 plate IDs stored in memory If an 11th Plate ID is added it will overwrite the first Plate ID stored in memory NOTE If ithe internal barcode Scanner option is installed the reader will automatically scan the plate barcode label and use this as the Plate ID P...

Page 112: ...Figure 3 67 Enter Sample ID screen Use the KEYPAD to enter numbers and the LEFT RIGHT arrow keys to move the cursor Prompts for Well Location Well locations Figure 3 68 can be changed at run time if a Manual Map has been specified and the user has requested a sample count at run time via the Utilities menu S A M P 1 L O C A T I O N R E P 1 G 0 1 Figure 3 68 Enter Well Location screen Use the KEYBO...

Page 113: ... E I N C A R R I E R A N D P R E S S R E A D K E Y Figure 3 69 Plate and Read prompts Press the READ key to initiate the plate read After the plate has been read all requested reports will be generated To halt in read in progress press the STOP key iii NOTE If using the incubation option the reader will wait for the incubator to reach temperature before reading the plate ...

Page 114: ...S A Y L I S T Figure 3 71 Print Report Menu Select the RESULT option to obtain an exact copy 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 Select MAP to provide a location matrix of the Blanks Standards Controls and Samples for a selected assay Select ASSAY to provide...

Page 115: ...f YES is selected for this screen you will be able to edit the replicates for each standard If NO is selected the user will be asked if reports should be printed You will see this screen only if you have a standard curve for this assay and have selected YES for Edit Standard Outliers E D I T S T D 1 R E P 1 Y E S Y E S N O Figure 3 74 Edit Replicate screen If YES was previously selected you will b...

Page 116: ... of programmed well locations in the selected geometry Assay Select REPORT at the Main Menu then select ASSAY S E L E C T A S S A Y N U M B E R 0 1 N A M E H B S A G Figure 3 77 Select Assay screen 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 Select R...

Page 117: ...icroplate covers when not reading the plate Although the PowerWave works with all flat U bottom and V bottom microplates it performs best with optically clear flat bottomed wells Non uniformity in the optical density of the well bottoms can cause loss of accuracy especially with U and V bottom polyvinyl microplates Check for this by reading an empty microplate Dual wavelength readings can eliminat...

Page 118: ...at Bio Tek prior to shipment and should operate properly upon initial setup If it is suspected that problems may have occurred during shipment if the device was reshipped by you or if regulatory requirements dictate that Installation Qualification Testing is necessary the following tests should be performed After the initial confirmation of operation this testing should be performed annually Syste...

Page 119: ... Calibration Test Plate provides a multiband calibration filter in location C6 The filter is Didymium glass V10 The filter should be scanned between 580 and 590 nm in 1 nm increments using KC4 For PowerWave 340 or if KC4 is not available select six wavelengths at 1 nm increments near the expected peak The wavelength of the maximum absorbance should be compared with the wavelength written on the sh...

Page 120: ...n Routine Procedure To ensure proper operation of PowerWave on an ongoing basis the following Operational Qualification Procedure should be conducted monthly System Self Test described above Calibration Test Plate described above ...

Page 121: ...this chapter we will discuss the TESTS utility The soft keys positioned under the menu options allow you to select the option 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 Figure 4 1 Selecting Tests from the Utility Options screen SOFT KEY 1 Press this key to select TESTS Use the MAIN MENU and PREVIOUS SCREEN keys to return to the Main Screen or move backwards throu...

Page 122: ...ument s System Self Test and reports results in a pass fail format The results are sent to the printer via the instrument s parallel port and a printout similar to Figure 4 3 is produced Photodiodes The Optics portion of the System Self Test confirms that the eight reading and 1 reference channels have adequate range gain within operational limits by comparison of the ambient readings to air in or...

Page 123: ...1 2 3 4 5 6 7 8 Air 13888 37248 38931 36215 38675 39845 34691 38657 39729 Dark 3737 3735 3751 3749 3734 3753 3742 3747 3737 Delta 10151 33513 35180 32466 34941 36092 30949 34910 35992 Lambda 547 Gain 1 68 Resets 4 Channel Ref 1 2 3 4 5 6 7 8 Air 13677 37188 38765 36129 38326 39678 34693 38571 39682 Dark 3736 3733 3748 3746 3732 3751 3740 3745 3735 Delta 9941 33455 35017 32383 34594 35927 30953 348...

Page 124: ...FT KEY 2 is selected the software s checksum appears on the display as shown in the example below Software P N Software Version Code Checksum 1ED3 dependent on software version The second screen shows Configuration P N Configuration Version Blank ...

Page 125: ... Plate Test This test is used to confirm the alignment repeatability accuracy linearity of the PowerWave An alternative method used to determine accuracy repeatability and linearity is the Linearity or Wet Test described later in this chapter The Calibration Test Plate PN 7260522 measures a known optical standard Both accuracy linearity repeatability and alignment are tested Specific calibration v...

Page 126: ... Utility Options menu select SETUP by pressing the soft key beneath this menu option 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 Figure 4 6 Accessing Setup From the SETUP options menu press the soft key beneath MORE to advance to the EDIT SETUP menu that allows entry of Calibration Plate values 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 L A M B D A M ...

Page 127: ...TER key to advance to the menu that allows entry of Well Location and calibration values W A V E L E N G T H 4 0 5 W E L L C 0 1 C A L I B R A T I O N V A L U E S 0 0 0 0 Figure 4 9 Entering Calibration Values using the Cal Plate Insert After each entry press ENTER to advance to the next consecutive well location Enter the values listed on the Calibration insert packaged with the plate The cursor ...

Page 128: ...P L T E Figure 4 10 Selecting the Calibration Plate Test Select the appropriate wavelength at the CALIBRATION FILTER screen by pressing the soft key beneath the desired value Press ENTER to save the value and advance to the next screen C A L I B R A T I O N L A M B D A 4 0 5 4 0 5 4 5 0 4 9 0 6 3 0 Figure 4 11 The Calibration Filter Selection screen When prompted insert the calibration plate into ...

Page 129: ...alibration Test Plate for more information If you have any questions about using your Bio Tek Universal Calibration Test Plate with your PowerWave on board Test Software or any of the test results indicate FAIL please contact Bio Tek s Technical Assistance Center at 800 242 4685 ...

Page 130: ...y of Calibration Plate wells C01 D04 E02 F05 G03 and H06 with known standard values contained in the Specification Sheet that accompanies each Calibration plate If the accuracy specifications are met then the reader also proves to be linear Repeatability This parameter is a measure of the instrument s ability to read the same well with minimum variation between two reads with the well in the same ...

Page 131: ...inations are given in Table 4 1 Table 4 1 Typical Enzyme Substrate Combinations and Stopping Solutions Enzyme Substrate Stopping Solution Alkaline Phosphate o nitrophenyl phosphate 3N sodium hydroxide beta Galactosidase o nitrophenyl beta D galactopyranoside 1M sodium carbonate Peroxidase ABTS 2 2 Amino di 3 ethyl benzthazolin sulfonate citrate phosphate buffer pH 2 8 Peroxidase o phenylenediamine...

Page 132: ...all the color is spread throughout the solution This is entirely a trial and error process If it does not give enough absorbance add another drop of food coloring The reader will display absorbance units up to 3 000 absorbance units it is specified to be linear to 2 000 absorbance units This is a configurable value and may vary depending on the reader model 4 Set up a rack containing 10 tubes numb...

Page 133: ...rsus percentage You should get a straight line passing through zero if your pipetting is accurate 8 The choice of dilutions is strictly arbitrary as is the absorbance of the original solution Use Table 4 2 as a model for calculating the expected absorbances of a series of dilutions given a different absorbance of the original solution Verification of Alignment Use a new microplate free from dust a...

Page 134: ...ure described in the Repeatability section 2 Compare the Means from each well using the readings in the two positions The readings should be within 1 of the OD and 0 010 counts of each other For example if A1 was initially 1 000 then the turn around test should show results between 980 OD and 1 020 OD Repeatability specification 1 005abs 405nm to 2 00abs 1 Read the microplate using a single wavele...

Page 135: ...al Considerations Any laboratory instrument which has been used for clinical analysis is considered a biohazard and should be decontaminated prior to handling Intact skin is generally considered an effective barrier against infectious organisms however small abrasions and cuts may not be always be visible Prophylactic gloves must be worn when handling instruments that have not been decontaminated ...

Page 136: ...household bleach a 6 1 mixture is required This is a caustic solution It is important to wear gloves and eye protection when handling it Wipe down the carrier and all exposed surfaces of the unit with the bleach solution Remove the top shroud of the instrument and wipe down the top surface of the instrument base as well as the inside of the top shroud Reassemble the unit and discard the used glove...

Page 137: ... under computer control For spectrum scanning and multiple wavelength readings PowerWave must be controlled through KC4 The RS 232 Port At baud rates of 1200 2400 and 9600 PowerWave is capable of sending and receiving data through its serial port RS 232C Controlling the Reader with Bio Tek Software PowerWave can be controlled for endpoint or kinetic single or dual wavelength readings using Bio Tek...

Page 138: ... is configured as a DTE that is the unit is wired to emulate a modem Data is received on Pin 2 the RX Pin and transmitted on Pin 3 the TX pin For an illustration see the section Setting Up the Serial Port for Communications in Chapter 2 The transmitted data consist of 96 six character strings Each string consists of a comma a sign character and four digits that represent an optical density reading...

Page 139: ... a blank well N Negative value used as a blank well O Out of range value used as a blank well space Output value has not been read Table B 1 Sign Characters When an asterisk or an O character is used the four digits that follow are undefined and ignored by the receiving program although they should be ASCII printable characters The space character should be followed by four more space characters ...

Page 140: ... No Parity 8 Data Bits and 2 Stop bits are fixed and cannot be changed Refer to Table B 1 for the ASCII control characters used in the computer control protocol and to Table B 2 for the ASCII characters used for PowerWave s buttons and keys USING THE STOP KEY TO HALT SERIAL COMMUNICATIONS Pressing the STOP key on the reader during serial communications will cause the following to occur 1 Halt the ...

Page 141: ...knowledge 06 06 F NAK Negative acknowledge 15 21 U RS Record separator 1E 30 ETX End of text 03 03 C DLE Data link escape 10 16 P CR Carriage return 0D 13 M LF Line feed 0A 10 J CTRL Z Control Z 1A 26 Z Table B 2 ASCII Control Characters Used in Computer Control Protocol All commands can be executed from the serial port As the final part of executing all commands PowerWave transmits a status strin...

Page 142: ... format of these files see Table B 4 W Upload the filter wavelength setup table to the computer by sending 6 three character strings separated by commas These strings correspond to the wavelengths assigned to lambda numbers 1 through 6 for example 405 450 490 630 000 000 A one byte command that returns 24 bytes of data before the status X Simulate pressing the STOP key on the reader This one byte ...

Page 143: ...ription Set temperature Get temperature Spectral scan see page B 20 for details Instrument identification see page B 16 for details Strip read mode see page B 16 for details Get minimum kinetic intervals see page B 24 for details Table B 3 Computer Control Command Set Cont ...

Page 144: ...s structure 2 7 NOT USED 8 29 NOT USED 30 Options Bit Set to 1 Bit Set to 0 Bit 6 dual wavelength single wavelength 31 Options Bit Set to 1 Bit Set to 0 Bit 4 NOT USED Bit 5 NOT USED Bit 6 NOT USED Bit 7 TTL minutes number of reads 32 NOT USED 33 34 NOT USED 35 36 NOT USED 37 NOT USED 38 45 NOT USED 46 49 NOT USED 50 52 Filter 1 ASCII filter Table B 4 Byte Assignment for Parameter Setup File ...

Page 145: ...ds on dual wavelength 62 63 Repeated reads count 1 9999 64 Map number 0 5 0 user constant 1 full blank 2 half blank 3 quarter blank 4 8 rows blank 5 12 columns blank 65 66 User blank constant value 0 000 to 2 999 67 162 Blanking map If constant blank is being used all the wells would be 00000000 sector number 1 12 Bits 0 3 unused Bit 4 unused Bit 5 1 suppresses the reading 0 enables the reading Ta...

Page 146: ...well as a blank well Bit 7 163 165 Not used 166 167 Length of read in minutes for TTL minutes in Repeated Reads range is 1 to 9999 minutes Bit Set to 1 Bit Set to 0 168 Delay in Read No Delay in Read Bit 0 169 170 Length of delay in seconds 0 to 999 Table B 4 Byte Assignment for Parameter Setup File Cont ...

Page 147: ...us string S3 A hexadecimal digit 0 through F where the bits mean 8 stepper motor busy 4 printer busy 2 1 S2 A single digit used as a reader fault or error code number ERROR CODES Error code numbers indicate the following 0 no fault or error 3 analog channel out of range 6 printer busy for more than 3 seconds 7 checksum byte of downloaded file does not match 8 WCF error detected during plate read S...

Page 148: ...r an EL312e The multiple byte strings such as Array 33 34 and Array 35 36 are listed as low byte first and high byte second This is how the reader expects the strings to come across Array 1 Loop through each byte of the following array starting at 2 Put that number in Array 1 This is a checksum for this file structure Comments and Order Array 2 7 NOT USED Array 8 29 NOT USED Array 30 0 Array 31 0 ...

Page 149: ...igh byte is the order Array 62 63 0 0 low byte high byte is the order Array 64 1 Full blank map Range is 0 to 5 for the blank maps Array 65 66 0 No user blank constant 0 000 to 3 000 to be entered Fixed point integer low byte high byte Array 67 162 Each well is given an 8 bit designation EXAMPLE Part 1 Array 67 could be 10000001 which is 81h ...

Page 150: ...ing well One 1 selects the well as a blanking well 0 selects it as a non blanking well In the above case this well is in Sector 1 the reading is enabled and it is used as a blank well The first 8 bit number is A 1 the second A 2 the thirteenth B 1 etc EXAMPLE Part 2 Array 68 162 00000001 which is 1h This means that all other wells are not used as blanking wells and they all belong to Sector 1 If t...

Page 151: ...ppropriate command from the asynchronous computing package used Cmdstr V 170byte array 2 Check for Status 000 which is an automatic return message from the reader 3 Start the reader Send CmdStr S 4 The program has to wait for the data to come across 650 characters plus 5 bytes of status are expected ...

Page 152: ...ill response with a single 1 All other instruments will return NAK STRIP READS The Select Strips command is defined as follows Host Limits Response ACK NAK first strip 1 0 12 first strip to read last strip 1 0 12 last strip to read Example 0x09 0x0C means read strips 9 through 12 The Select Strips command returns the standard status response string To read a range of one or more contiguous strips ...

Page 153: ...Data returned for each set of strips read at an indicated wavelength will start with CR followed by that wavelength s well ODs as defined in the previous section Each set of strips will be terminated with a Z character followed by a checksum Checksum calculation starts with the first byte after the data start code CR up through and including the Z data terminator code The checksum is transmitted a...

Page 154: ...der V command d If reading a range of selected strip s send Select Strips command as defined above e Send Read command S to perform the actual read Response data from the read will consist of an entire 8 x 12 plate for each indicated wavelength or an 8 x strips plate consisting of strip s selected by the Select Strips command Wavelengths indicated by 000 will return no data NOTE Data returned for ...

Page 155: ...e beeping Otherwise another Z character will be sent This indicates that the reader is completely finished with the read and that more commands can be sent by the host Since the command function described above is new there is no reason to maintain compatibility with obsolete products Therefore no assay name report title or time date stamp blank or otherwise will be returned with data collected us...

Page 156: ...us response string After wavelength configuration a self test command sequence must be run to calibrate gains and generate self check information for the new wavelength configuration SPECTRAL SCANNING The Spectral Scan command is defined as follows Host Limits Response ACK NAK well 2 00 scan entire strip 01 08 scan well in indicated row strip 2 01 12 start wave 3 200 800 stop wave 3 200 800 wave s...

Page 157: ...t the spectral scan has terminated and no further response data will be transmitted by the reader Reader response protocol ASCII format for each wavelength undergoing calibration none to all wavelengths selected wavelength 3 200 800 wave terminator 1 CR calibration terminator 1 Z always returned even if no waves calibrated For each wavelength none or all wavelengths selected wavelength 3 200 800 w...

Page 158: ...s returned alone with no wavelengths If a single well is selected for scanning or if last scan is selected for a series option no first strip read is performed In this case the Z terminator is returned alone with no wavelengths Checksum calculation starts with the first byte AFTER the data start code up through and including the Z data terminator code The checksum is transmitted as one hex data by...

Page 159: ...s been detected 0x00 0x00 hex not ASCII will be returned as temperature data An 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 should be able to accept and process the Get Temperature command at any time including during a read cycle The temperature response ...

Page 160: ...onse string Intervals are returned in units of seconds Minimum kinetic intervals are calculated based on a number of different variables including but not limited to the user s selection of strip s wavelength s shake time and baud rate The host PC should ensure that all reader setup commands have been sent for the upcoming read before requesting the minimum kinetic interval filter configuration as...

Page 161: ...which means the instrument will still respond to keypad input An eight in the first digit denotes a serious error which requires that the reader be powered down before additional keystrokes will be accepted 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 ERROR 0202 Monochroma...

Page 162: ...substantial noise 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 Auto Cal sequence PROBABLE CAUSE Carrier A loose belt lo...

Page 163: ...o its home position in too few or too many steps the test fails PROBABLE CAUSE The belt is slipping due to incorrect tension a loose motor pulley or a loose belt clamp This can also be caused by a bad motor drive circuit Displayed Error Potential Cause ERROR 0500 Saturation error Error 500 indicates an empty filter location on the filter wheel or that the filter wheel is not installed PROBABLE CAU...

Page 164: ...ance to specifications PROBABLE CAUSE A bad lamp could provide enough reduction in signal to cause this error Misaligned optics could also cause this error Displayed Error Potential Cause ERROR 0700 Reader failed noise test Error 700 indicates that the measurement electronics do not read repeatably at a maximum gain with the light blocked PROBABLE CAUSE External signals penetrating the measurement...

Page 165: ...lue 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 The measurement electronics background noise has changed since the last power up self check Displayed Error Potential Cause ERROR 0A00 Read time air blank out of range Error A00 indicates that the blank f...

Page 166: ...parallel port may not be correctly selected Displayed Error Potential Cause ERROR 0D00 Reader failed calibration checksum test ERROR 0E00 Wavelength not detected in reader s wavelength table 1 MEAS 2 Ref 3 sec ERROR 0F00 Reader s wavelength or channel signal is out of specified range ERROR 1000 Required reader configuration data missing ERROR 1100 Failed configuration checksum test ERROR 1200 Cali...

Page 167: ...ter control assay definition error ERROR 1700 Kinetic interval too short for selected options ERROR 1800 Too many kinetic intervals selected ERROR 1900 Memory allocation failure These errors indicate the reader has failed one or more internal self tests Contact Bio Tek s Technical Assistance Center for instructions ...

Page 168: ...C 8 Error Codes ...

Page 169: ...ave The following are examples of reports that can be generated and or printed from the PowerWave Refer to Chapter 2 Output Options for details on how to select to print these reports In addition an Assay List Assay Definition Map and Result can be printed by choosing Report from the Main Menu screen ...

Page 170: ...D 2 Report Format Figure 1 Samples with Calls on Matrix Report ...

Page 171: ...Appendix D D 3 Figure 2 Curve Fit Report ...

Page 172: ...D 4 Report Format Figure 3 Samples with Calls on Column Report ...

Page 173: ...Appendix D D 5 Figure 4 Column Report without Samples ...

Page 174: ...D 6 Report Format Figure 5 Panel Report ...

Page 175: ...Appendix D D 7 Figure 6 Assay Detail Report ...

Page 176: ...D 8 Report Format Figure 7 Assay List ...

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