Page 1 PowerWave Microplate Scanning Spectrophotometer Operator’s Manual...
Page 3 Manual Upate PowerWave Rev. C1, P/N 7261000 The following information pertains to revised instructions for unpacking and repackaging the reader as shown in Figure 2-1, Chapter 2, page 2-2. Figure 2-1 illustrates how to unpack the reader. Figure 2-1: Unpacking and repackaging the reader •...
Page 7 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.
Page 8 Revision Appendix Revision Date Chapter Change 4/97 First Release 4/97 Front Pg Changed Enzyme Immunoassay Analyzer to Microplate Spectrophotometer. Changed ELISA-based data to microplate-based data. Changed all references to monochronometer to monochromator. 8/97 Inserted a Manual Update correcting the calibration plate Part Number.
Page 9: Warnings And Precautions
Warnings and Precautions The PowerWave should be operated on a flat surface away from direct sunlight or strong incandescent light. Excessive humidity should be avoided. Hazard Warnings Warning! Power Rating. PowerWave’s must be connected to a power receptacle that provides voltage and current within the specified rating for the system.
Page 10 Precautions The following precautions are provided to help you avoid damaging the system: Caution: Service. The system should be serviced by authorized service personnel. Only qualified technical personnel should perform troubleshooting and service procedures on internal components. Caution: Environmental Conditions. Do not expose the system to temperature extremes.
Page 11 Electromagnetic Interference and Susceptibility USA FCC CLASS A Warning: Changes or modifications to this unit not expressly approved by the manufacturer could void the user's authority to operate the equipment. This equipment has been tested and found to comply with the limits for a Class A digital device, pursuant to Part 15 of the FCC Rules.
Page 12 Based on the testing below, PowerWave bears the CE mark. EC DIRECTIVE 89/336/EEC EN 50081-1, CLASS A-Emissions The system has been type tested by an independent testing laboratory and found to meet the requirements of EC Directive 89/336/EEC for Radiated Emissions and Line Conducted Emissions.
Page 13 Safety Symbols The following warning and informational symbols may be found in various locations on the PowerWave. Only qualified personnel who recognize shock hazards and are familiar with the safety precautions should use this instrument. Read the manual carefully before operating this instrument.
Page 14 Off (Supply) Arrest (alimentation) Aus (Trennung vom Netz) Desconectado Aperto (sconnessione dalla rete di alimentazione) Caution (refer to accompanying documents) Attention (voir documents 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...
Page 15: Intended Use Statement
ELISA test kit, in accordance with the test kit's procedure. A versatile curve 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.
Page 16 About This Manual The intent of this Operator’s Manual is to quickly instruct the new user how to set up and operate Bio-Tek’s PowerWave. To help you read and understand this manual, certain document conventions have been used. Important information will be flagged in this manner, with the information icon that appears at the beginning of this paragraph.
Page 17 Warranty This Warranty is limited and applies only to new products, except for computer-based software which is covered under a separate Warranty Policy, manufactured by Bio-Tek Instruments, Inc. (“Bio-Tek”). Bio-Tek makes no warranty whatsoever regarding the condition of used products. Bio-Tek warrants the instrument (hereinafter collectively referred to as “Products”...
Page 19: Table Of Contents
Table of Contents Notices ..................... Warnings and Precautions .............. Intended Use Statement ..............Chapter 1 Introduction..............1-1 Introducing the PowerWave ............... 1-1 Quality Control ..................1-2 Hardware Features ................1-3 Software Features................1-3 Package Contents................1-4 Optional Accessories ................1-4 Specifications..................1-5 Technical Support ................
Page 20 Adjusting the Fuse Configuration or Fuse Replacement ......2-15 Other Utility Options ..................2-17 Setup Options ................... 2-17 Output Options.................. 2-19 Read Options ..................2-20 Chapter 3: Operation..............3-1 Introduction ...................3-1 The Keyboard ..................3-1 The Cooling Fan ................. 3-2 The Startup Screen ..................3-2 The Main Menu Screen ...................
Page 21 Number of Kinetic Points Selection ..........3-15 Onset OD Selection..............3-15 Well Scanning ................3-16 Map Definition....................3-17 Map Generation................3-18 Mapping Direction ............... 3-19 Replication Direction ..............3-19 Start Mapping at Well Location ........... 3-22 Blank Map Selection..............3-22 Constant Blank Value..............
Page 22 Formula Entry................3-42 Validation Formula Examples............3-44 Number of Required Controls/Blanks.......... 3-45 Cutoff Formulas................3-45 Grayzone..................3-46 Positive or Negative Call for Cutoff ..........3-46 Transformations ................3-47 Transformation Formula Definition ..........3-47 Transformation Scope Variable........... 3-48 Defining the Curve-Fit................... 3-52 Curve-Fit..................
Page 23 Chapter 4: Performance Verification ..........4-1 Achieving Maximum Performance..............4-1 Installation and Operational Qualification Procedures........ 4-2 Routine Procedure ..................4-4 Accessing Tests and Calibration Using the Utility Option ......4-5 Test Options ....................4-5 System Self Test ................. 4-6 Checksum ................... 4-8 Calibration Validation ..................
Page 25: Introducing The Powerwave
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.
Page 26: Quality Control
Bio-Tek’s PowerWave may be configured with all, or selected options for optimum performance: • PowerWave 340 instruments are capable of reading plates at wavelengths between 340 and 800 nm. • I-model instruments have a four-zone incubation chamber which controls temperature from 4° above ambient to 50° C.
Page 27: Hardware Features
• 55 assays are available onboard; 75 assays can be pre-programmed and exported to the reader using Bio-Tek’s Extensions software. • Automatically stores results for last 10 plates. • Spectral scanning on PowerWave 200 using KC4 software. PowerWave Operator’s Manual...
Page 28: Package Contents
Package Contents _______________________________________________ The contents of the PowerWave 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) •...
Page 29: Specifications
200 to 800 nm for PowerWave 200 340 to 800 nm for PowerWave 340 λ accuracy: +/- 2 nm for PowerWave 200 and 340 λ repeatability: +/- 0.2 nm for PowerWave 200 and 340 Regular read mode end point or kinetic with interval at least 20 seconds.
Page 30 Repeatability (STD): +/- 1% +/- 0.005 Abs from 0 to 2.000 Abs +/- 3% +/- 0.005 from 2.000 to 2.5 Abs (measured @ 405 nm) • Display: 2 X 24 Character LCD • Light Source: Xenon Arc Flash bulb • Dimensions: 16.0”...
Page 31 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. PowerWave Operator’s Manual...
Page 32: Technical Support
Technical Support ____________________________________________________________________________________________________________________________________ Bio-Tek’s PowerWave is backed by a superior support staff. If the Microplate Reader ever fails to work perfectly, please contact the Technical Support Staff. • You can telephone the Technical Assistance Center between 8:30 AM and 6:00 PM Eastern Standard Time, Monday through Friday, except Holidays.
Page 33: The Operating Environment
The Operating Environment For best operation, install Bio-Tek’s PowerWave on a level surface in an area where ambient temperatures between 18°C (65°F) and 40°C (104°F) can be maintained. The reader is sensitive to extreme environmental conditions. Conditions to avoid are: •...
Page 34 with improper packaging materials may void your warranty. If the original packing materials have been damaged, replacements are available from the manufacturer. • An RMA (Return Materials Authorization) must be obtained before returning equipment for service. Contact Bio-Tek’s Technical Assistance Center at 1-800-242-4685 for an RMA number, and mark the RMA number on the outside of the shipping container.
Page 35: After Unpacking, Verify Performance
After Unpacking, Verify Performance Each time the PowerWave is turned on, it automatically runs a self-test. If an error occurs, contact Bio-Tek. Before Repackaging the Instrument: • Decontaminate the reader prior to shipping. (Refer to the Decontamination procedure in Appendix A.) •...
Page 36: Adjusting The Reader's Wavelength Table Setting
Adjusting the Reader’s Wavelength Table Setting To check, or change the software wavelength: • Power up the reader. At the Main Menu Screen (Figure 2-2), press the soft key under the UTIL menu option. R E A D Y 9 : 4 5 P M R E A D D E F I N E R E P O R T...
Page 37 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. PowerWave Operator’s Manual...
Page 38: Printing And Data Communications
Printing and Data Communications _____________________________ Installing a Printer The PowerWave’s parallel port (LPT1) (see Figure 2-7) allows connection to Epson-compatible printers such as the Canon Bubble Jet. The Bubble Jet printer should be configured for Epson graphics compatibility. The reader’s parallel port requires a 25-pin D-sub connector.
Page 39: Parallel Port Pin Definition
Make sure the securing screws on both ends of the cable are tightened, and power up the reader and printer. 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)
Page 40: Setting Up The Serial Port Communications With Other Devices
Setting Up the Serial Port for Communications with Other Devices The PowerWave has a 25-pin serial (RS232) port located on the rear panel of the instrument (see Figure 2-7). The serial port allows the reader to communicate with a computer, using standard communications software and/or RS232 protocols.
Page 41: Serial Port Pinout Description
Signal Table 2-2: Serial Pinout Description Setting Up Communications 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: •...
Page 42: Changing The Baud Rate On The Reader
Changing the Baud Rate on the Reader To change the baud rate from the default of 2400, to either 1200 or 9600: • Power up the instrument. At the Main Menu screen (Figure 2-8), press the soft key beneath the menu option UTIL. R E A D Y 9 : 4 5 P M R E A D...
Page 43 S E T T I N G S : N O P A R I T Y S T O P - B I T S D A T A - B I T S Figure 2-12: Viewing Other Communication Settings PowerWave Operator’s Manual 2-11...
Page 44: Lamp Replacement
Lamp Replacement _______________________________________________ The reader’s lamp should operate without replacement for about 1 billion flashes. The intensity of the lamp will slowly drop over time until the reader’s run time self check detects a low signal, and warns the user with a displayed message.
Page 45: Bulb Removal
Bulb Removal • Always wear safety glasses when handling the bulb. • Turn the PowerWave off and let it sit for 5 minutes before proceeding. • Remove the bottom plate mounting hardware (6). • Remove the bulb socket mounting hardware (2) and carefully remove the bulb housing.
Page 46: Adjusting The Line Voltage Input Range
Adjusting the Line Voltage Input Range The PowerWave is equipped with a four-voltage range power input module. This power input module, located on the right side of the instrument, can be adjusted for 100 VAC, 120 VAC, 230 VAC and 240 VAC voltage inputs.
Page 47: Adjusting The Fuse Configuration Or Fuse Replacement
Contact Technical Services if the fuse replacement fails to rectify the problem. • Use a small, flat-blade screw driver to remove the fuse module from the power input module. PowerWave Operator’s Manual 2-15...
Page 48 • The fuse module has two fuse configurations: The USA configuration has a fused Hot (PN 46023) .75 amp slo blo). The European configuration has both Hot and Neutral fused (PN 46051) .315 amp slo blo T 5 X 20 mm. •...
Page 49: Other Utility Options
Other Utility Options ______________________________________________ The PowerWave may be configured a number of ways, depending on user preference. Configuration options are accessed via the Utility Options menu (Figure 2-15), and include: • SETUP: Setting the date and time. • OUTPUT: Selecting whether reports will be output to a Printer, the Computer display or both;...
Page 50 • Enter the new date, using the NUMERIC keys. The cursor is positioned under the first editable field, and advances automatically. To change the date format , press the soft key beneath MMDDYY or DDMMYY. The display updates to reflect the new format (Figure 2-17).
Page 51: Output Options
Figure 2-21: Choosing Whether to Include samples in Column Report • If NO is selected, the column report will only include standard blank and control well results. • Press ENTER to advance to the PRINT CURVE FIT screen (Figure 2-22). PowerWave Operator’s Manual 2-19...
Page 52: Read Options
P R I N T C U R V E - F I T ? Y E S Figure 2-22: Choosing Whether to Print Curve Fit data • Any previously defined selection is displayed on the top line of the screen. To change the report option, press the soft key under YES or NO.
Page 53: Chapter 3: Operation
PowerWave, and its software. Introduction The PowerWave features a 25-pad keyboard and a 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.
Page 54: The Cooling Fan
The Cooling Fan Located on the bottom right edge of the instrument is the switch which activates the cooling fan. When the PowerWave is on, the temperature in the measurement chamber will be approximately 7°C above ambient temperature. For users that need the measurement chamber to be close to ambient, Bio-Tek recommends using the cooling fan.
Page 55: The Main Menu Screen
REPORT option: Allows printing of results and protocol descriptions. The user will be prompted for the name of a previously run assay with valid data. • UTIL option: The user will be prompted to select options from the list of onboard utilities. PowerWave Operator’s Manual...
Page 56: Define
Define The Main Menu option, DEFINE, allows the user to edit previously defined assays stored in memory. • From the Main Menu, press the soft key beneath the DEFINE menu option to access the SELECT ASSAY NUMBER screen. Select Assay At the Select Assay Number screen, •...
Page 57: Assay Name
The cursor will wrap around the edit field. • Use SOFT KEYS 1, 2, 3 & 4 when using alphanumeric characters on the display above the soft key in the assay name. PowerWave Operator’s Manual...
Page 58: Define (Method, Map, Formula And Curve)
Define (Method, Map, Formula and Curve) The DEFINE Option screen (Figure 3-5) allows the user to edit the Method, Map, Formula or Curve Fit. 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 3-5: Define Option screen Method...
Page 59: Delay In First Read
Figure 3-7: Delay in first read entry • Enter the time in minutes and seconds, using the numeric keys. • Use Main Menu to save the time value entered. • Use Previous Screen save the time value entered. PowerWave Operator’s Manual...
Page 60: Incubation Temperature
INCUBATION TEMPERATURE ENTRY OPTIONAL FEATURE The incubation temperature screen (Figure 3-8) allows the user to set the assay incubation temperature. I N C U B A T I O N T E M P : 3 7 C A M B I E N T T E M P E R A T U R E Figure 3-8: Incubation Temperature entry •...
Page 61: Single Or Dual Wavelength
Press SOFT KEY 1 to select SINGLE wavelength. The top line of the display is updated to reflect the selection. • Press SOFT KEY 2 to select DUAL wavelength. The top line of the display is updated to reflect the selection. PowerWave Operator’s Manual...
Page 62: Wavelength Selection
WAVELENGTH SELECTION The MEAS selection screen (Figure 3-10) allows the user to select the wavelength(s) for the assay. Any previously defined wavelength appears on the top line of the display and the options on the second. M E A S : 4 5 0 R E F : 6 3 0 4 0 5 4 5 0...
Page 63: Kinetic Interval
= 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 numeric entry fields. PowerWave Operator’s Manual 3-11...
Page 64: Kinetic Number Of Reads
KINETIC NUMBER OF READS ENTRY Use this screen (Figure 3-13) to enter the number of kinetic reads. K I N E T I C T O T A L N U M B E R R E A D S : Figure 3-13: Kinetic Reads entry screen •...
Page 65: Shake Time
S H A K E S P E E D : M E D I U M L O W M E D I U M H I G H V A R I Figure 3-17: Shake Speed Selection screen PowerWave Operator’s Manual 3-13...
Page 66: Kinetic Data Analysis Selection
• Use SOFT KEY 1 to select low-speed (17 Hz) shaking. • Use SOFT KEY 2 to select medium-speed (18 Hz) shaking. • Use SOFT KEY 3 to select high-speed (19 Hz) shaking. • Use SOFT KEY 4 to select variable-speed shaking ( 1 second of each speed repeated).
Page 67: Number Of Kinetic Points Selection
Figure 3-20: Onset OD screen • Use NUMERIC keys to enter the onset OD. 3.000 Abs is the maximum value. • Use the LEFT and RIGHT ARROW keys to move the cursor within the entered OD field. PowerWave Operator’s Manual 3-15...
Page 68: Well Scanning
WELL SCANNING If Scanning is chosen as the Read Type, use the following screen (Figure 3-21) to enter the number of scan points per well: E N T E R N U M B E R S C A N P O I N T S ? Figure 3-21: Number of Scans screen The maximum number of selectable points is 31 (odd number only).
Page 69: Map Definition
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: Selecting the Map option on the DEFINE screen PowerWave Operator’s Manual 3-17...
Page 70 • At the DEFINE Options screen, press Soft Key 2 to begin the plate MAP process. MAP GENERATION This option (Figure 3-23) allows the user to select manual or automatic plate map generation for the assay. Any previously defined value is shown on the top line of the display and the available options on the second.
Page 71 R E P D I R E C T I O N ? A C R O S S D O W N A C R O S S Figure 3-25: Mapping the Direction of the Replicates PowerWave Operator’s Manual 3-19...
Page 72 • Press SOFT KEY 1 to map the replicates DOWN the column, following the direction 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.
Page 73 The map above shows Map Direction DOWN, Rep Direction ACROSS STD1 STD2 STD3 STD4 STD5 STD1 STD2 STD3 STD4 STD5 The map above shows Map Direction ACROSS, Rep Direction DOWN Figure 3-25A: Examples of Mapping Directions PowerWave Operator’s Manual 3-21...
Page 74 START MAPPING AT WELL LOCATION The Start Mapping at Well Location screen (Figure 3-26) is only shown if automatic mapping is selected. This option allows the user to enter the location of the well that will be the starting point for automatic mapping. Previously defined Starting Well Location appears on the top line of the display.
Page 75 COLUMN enables a single blank well or an average of blank wells to be selected for each column. The blank OD or average OD will be subtracted from other wells in the column. Use manual mapping to position blanks, standards, controls and samples. PowerWave Operator’s Manual 3-23...
Page 76: Constant Blank Value
• P-ACROSS enables a blank in every even-numbered column to be subtracted from the well to the left of it in every odd column. Use manual mapping to set up the appropriate map by placing the standards, controls and samples in only the odd columns. •...
Page 77: Number Of Blanks
Use the NUMERIC & ALPHA keys to enter a Blank Location, based upon the plate geometry. • Use the ARROW keys to move the cursor to the next or previous editable field. The cursor is positioned beneath the first editable field. PowerWave Operator’s Manual 3-25...
Page 78: Number Of Standards
NUMBER OF STANDARDS This option (Figure 3-31) allows the user to enter the number of standards that will be used in the assay. Any previously defined value will be displayed on the screen. If the number of standards is altered, the number of replicates for the standard automatically defaults to 1.
Page 79: Average Standards
Use the NUMERIC & ALPHA keys and the DECIMAL POINT key to enter Standard Concentration values. The range is .00001 to 999999, but the total field cannot exceed 6 characters for any concentration. Valid well locations for the defined geometry are listed below. PowerWave Operator’s Manual 3-27...
Page 80: Reuse Of Standard Curves
For example, enter A1 as A 0 1. REUSE OF STANDARD CURVES The PowerWave now has the ability to reuse a standard curve that has already been established (Figure 3-35). Limitations of the Reuse of Standard Curves ♦...
Page 81 If YES is chosen, a new standard curve will be generated. The plate map is not changed. (If “Prompt for Sample ID” is enabled in UTIL section, the user will be prompted to enter the number of samples - see page 2-20 for more information on the UTIL options.) PowerWave Operator’s Manual 3-29...
Page 82: Number Of Controls
If NO is chosen, the stored standard curve will be used. If Auto mapping had been used to originally map the standards, blanks, controls and samples defined for this assay, the map will be automatically regenerated without the standards, beginning in well xxx (where xxx was chosen as the Starting well in the map, usually well A01).
Page 83: Type Of Control
Any previously defined number will be displayed. N U M B E R R E P L I C A T E S : Figure 3-40: Number of Control Replicates screen PowerWave Operator’s Manual 3-31...
Page 84: Location Of Controls
• Use the NUMERIC keys to enter a value for Number of Control Replicates. The range is 1 to 12 replicates. The software performs a check to ensure the number of replicates, multiplied by the number of controls, does not exceed the number of undefined wells remaining on the plate.
Page 85: Number Of Samples
Any previously defined Sample Location appears on the display. S A M P # 1 L O C A T I O N R E P # 1 : A 0 Figure 3-44: Sample Location screen PowerWave Operator’s Manual 3-33...
Page 86 • Use the NUMERIC, ALPHA keys and DECIMAL POINT keys to enter the sample 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: ⇒...
Page 87: Panel
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. PowerWave Operator’s Manual 3-35...
Page 88 • 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 alphanumeric characters on the display above the soft key in the assay name.
Page 89 The OPTION key will increment the assay number by one. The display will be updated as the keys are pressed. • The NUMERIC keys are used to choose the assay numbers of the tests to be run. The range is 1 to 55. PowerWave Operator’s Manual 3-37...
Page 90 Once the first assay has been chosen, the software will prompt for the location that the reader should start mapping the assay (Figure 3-49). S T A R T M A P P I N G W E L L L O C A T I O N : Figure 3-49: Well location to begin mapping •...
Page 91 Figure 3-49A: Panel Report PowerWave Operator’s Manual 3-39...
Page 92: Formula Definition
Formula Definition Defining the Formula involves identifying the formula type and entering the actual formulas. Formula definition screens are displayed on the PowerWave in the order detailed below. Formulas created using Bio-Tek’s Extensions (Define Reader Protocol software) cannot be edited by using the reader.
Page 93: Formula Type
Figure 3-51: Validation Type selection screen • Press SOFT KEY 1 to select Control Validation Formula. • Press SOFT KEY 3 to select Assay Validation Formula. • Press SOFT KEY 4 to select Blank Validation Formula. PowerWave Operator’s Manual 3-41...
Page 94: Formula Entry
FORMULA ENTRY Use this screen (Figure 3-52) to enter the formula used in the assay. Each formula can contain a maximum of 24 characters. Spaces are unnecessary. F O R M U L A # 1 : M A T H O T H E R M A P F U N C T N...
Page 95 FUNCTION list: FUNCTION LIST LOG10 Log Base 10 ALOG10 Ante Log Base 10 Absolute Value Power ALOG Anti Log SQRT Square Root • Functions are followed by wells, well types or numbers in parentheses. PowerWave Operator’s Manual 3-43...
Page 96: Validation Formula Examples
VALIDATION FORMULA EXAMPLES • Blank Validation: An assay protocol states that the blank wells on a plate should have an OD of less than 0.050. The formula is entered on the reader as a Blank Validation Formula: BLK < 0.050. •...
Page 97: Number Of Required Controls/Blanks
All wells whose OD is higher than the mean of STD2 on a plate will be called POS. This formula would be entered as STD2;x in the Cutoff formula definition screen. PowerWave Operator’s Manual 3-45...
Page 98: Grayzone
NOTE: A syntax error in your formula will result in a “Token Error” message when running the assay. GRAYZONE ENTRY Use the Grayzone screen (Figure 3-54) to enter the grayzone percentage for the assay. The grayzone is the area around the cut-off, that is, samples that fall within an area defined by the cutoff (+/- 5%) would be considered equivocal or questionable.
Page 99: Transformations
Divide all OD’s on the plate by 2 and multiply by 100. Enter the formula: (OD/2)*100 This formula will be applied to the OD’s of all samples, standards, controls and blanks that are present on the plate map. PowerWave Operator’s Manual 3-47...
Page 100: Transformation Scope Variable
TRANSFORMATION SCOPE VARIABLE For more complicated transformations, a Transformation Scope Variable (TVar) can be defined. The TVar will allow the user to define a variable to be used in the transformation formula. This variable will define the scope of the transformation: whether to apply the transformation to just the samples (SMP) or to all wells defined on the plate (OD).
Page 101 SMP - NC;x To program this on the reader, • At the Scope Variable screen, select SMP. Press Enter. • Press TRANS. • Use the Math, and Map keys to enter the formula as shown above. PowerWave Operator’s Manual 3-49...
Page 102 If OD is chosen, the formula definition screen will appear. Use the formula keys (Math, Other, Map and Function) to define the transformation variable (TVar). Once the variable has been defined, it can be used in a transformation formula. The TVar will be available as a MAP option when writing the transformation formula.
Page 103 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. PowerWave Operator’s Manual 3-51...
Page 104: Defining The Curve-Fit
• Extrapolation of Unknowns These screens are displayed on the PowerWave in the order in which they appear in the assay. If a closed variable (i.e., an element of the assay definition that the user cannot access or modify) is being used in the assay, the entry screen is omitted.
Page 105 Logit/Log. Point to Point or “PT to PT”: A plot that connects each standard point with a line, with no averaging of the values to “smooth” the curve at each standard. PowerWave Operator’s Manual 3-53...
Page 106: Edit Standard Outliers
C U R V E - F I T T Y P E : C - S P L I N E N O N E L I N E A R Q U A D * M O R E C U R V E - F I T T Y P E : S P L I N E...
Page 107: Axis Selection
Press SOFT KEY 1 to select YES (extrapolate the unknowns). The top line of the display updates to reflect this selection. • Press SOFT KEY 2 to select NO. The top line of the display updates to reflect this selection. PowerWave Operator’s Manual 3-55...
Page 108 Note: If extrapolation is chosen for point to point curve fit, unknown concentrations will be extrapolated linearly from the nearest segment of the curve. If the plot includes both increasing and decreasing segments, the curve printout will be labeled “Ambiguous.” The resulting values, which actually are extrapolated, may not be indicated as such.
Page 109: Reading A Microplate
Figure 3-64: Assay Selection screen • Press ENTER to advance to the EDIT ASSAY NAME screen. • Labeled keys may be used to move around the reader’s menu structure, specify the assay number or clear the display: PowerWave Operator’s Manual 3-57...
Page 110: Run-Time Prompts
Run-Time Prompts After the assay is selected, the user may be prompted for information, depending on preferences selected in the reader’s Utilities, whether a manual map was set up, or if the assay was created in, and downloaded from, Bio- Tek’s Extensions...
Page 111: Enter Number Of Samples
Use the LEFT & RIGHT ARROW keys to move the cursor to the previous or next editable field. • Use SOFT KEYS 1, 2, 3 & 4 when using alphanumeric characters on the display in the field above the soft key. PowerWave Operator’s Manual 3-59...
Page 112: Enter Sample Id
Enter Sample ID If prompted, the user can enter a starting sample identification from 00001 to 99999 (Figure 3-67). The software will automatically increment each subsequent sample identification by 1. The sample IDs will be assigned by following the mapping order previously defined. E N T E R S A M P L E I D :...
Page 113: Beginning The Plate Read
• To halt in read in progress, press the STOP key. NOTE: If using the incubation option, the reader will wait for the incubator to reach temperature before reading the plate. PowerWave Operator’s Manual 3-61...
Page 114: Printing Reports And Assay Lists
Select ASSAY to provide a map and a listing of all of the assay’s settings, such as wavelengths, numbers of well types, all validation, cutoff, transformation, and general formulas and curve fit parameters. • Select LIST to provide a list of all assays in the PowerWave. 3-62 Operation...
Page 115: Result
Select YES if you wish to remove a replicate. P R I N T R E S U L T S ? Y E S Figure 3-75: Print Results screen • Press YES to compile reports, or NO to return to the Main menu. PowerWave Operator’s Manual 3-63...
Page 116: Map
Press ENTER to enter the assay and begin printing the map and other assay parameters. List • Select REPORT at the Main Menu, then select LIST. The entire list of assays stored in the PowerWave’s memory will be sent to the printer. 3-64 Operation...
Page 117: Chapter 4: Performance Verification
• 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.
Page 118: Installation And Operational Qualification Procedures
Tests outlined in this section may be utilized to confirm initial and ongoing performance of PowerWave. Your PowerWave reader was fully tested at Bio-Tek prior to shipment and should operate properly upon initial setup. If it is suspected that problems...
Page 119 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 sheet supplied with the calibration plate.
Page 120: Routine Procedure
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) Performance Verification...
Page 121: Accessing Tests And Calibration Using The Utility Option
Accessing Tests and Calibration Using the Utility Option From the Main Menu screen, press the soft key that corresponds to UTIL to access the PowerWave’s Utility menu. R E A D Y 4 5 P M R E A D...
Page 122: System Self Test
Main Screen or move backwards through the menu structure. System Self Test When SOFT KEY 1 is pressed, the PowerWave runs the instrument’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...
Page 123 Range: PASS Thermistor: PASS Zone 3: 31.3 Min: 31.3 Max: 31.3 Range: PASS Thermistor: PASS Zone 4: 29.9 Min: 29.9 Max: 29.9 Range: PASS Thermistor: PASS SYSTEM TEST PASS Figure 4-3: Output Results of the System Test PowerWave Operator’s Manual...
Page 124: Checksum
Checksum When SOFT 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) Performance Verification...
Page 125: Calibration Validation
Calibration Validation It is considered good laboratory practice to periodically validate the calibration of the PowerWave. Calibration validation should occur monthly by performing either of the tests in this section. The tests are: Calibration Plate Test Linearity Test (wet test) alternate method...
Page 126: Entering Calibration Plate Specifications
ENTERING CALIBRATION PLATE SPECIFICATIONS Using the Data Sheet provided with the Calibration Plate, enter the calibration values. To enter the values into the reader: From the Main Menu: R E A D Y 4 5 P M R E A D D E F I N E R E P O R T U T I...
Page 127 Enter the values listed on the Calibration insert packaged with the plate. The cursor will be positioned under the first editable field. • When all values have been entered, press the Main Menu key. PowerWave Operator’s Manual 4-11...
Page 128: Running The Calibration Plate Test
Running the Calibration Plate Test Power up the PowerWave. From the reader’s Main Menu display: • Press the soft key beneath UTIL to access the Utility Options menu. • At the SELECT UTILITY OPTION menu (Figure 4-4), press the soft key beneath TESTS.
Page 129 Please see the data sheet supplied with the Calibration 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 The Calibration Report has three report parameters: • Alignment: This portion of the test measures the alignment of the microplate carrier with the optical path. A reading of > .015 represents an out-of-alignment condition. Wells A01, A12, H08, and H12 are the only valid alignment holes for the reader on the 7260522 calibration test plate.
Page 131: Linearity Test
Yellow food coloring (Example: Durkee yellow food coloring, .3 oz. bottle)  • Flat-bottom microplates (Example: Costar #3590)  • Tween 20 (polyoxyethylenesorbitan Monolaurate) • Beakers • Rack of 10 test tubes (10 ml minimum volume) PowerWave Operator’s Manual 4-15...
Page 132: Procedure
PROCEDURE Create a 0.5% solution of Tween 20 and distilled water. As an example, add 1 ml of Tween 20 to 200 ml of distilled water. Shake well for several minutes. Add 6 drops of yellow food coloring to the 200 ml of the 0.5% Tween solution.
Page 133: Verification Of Alignment
• Use different solution volumes (not less than 100 microliters) and densities. For example, put 100 microliters in rows A through D, and 150 microliters in rows E through H. PowerWave Operator’s Manual 4-17...
Page 134 • The factory repeatability and accuracy tests that follow are  performed with Costar flat (#3590) and round bottom (#3797)  microplates with Durkee yellow food coloring, using a 405 nm filter. Different solutions and microplates, may vary repeatability and accuracy. Accuracy (specification +/-1%+/-.010) @ 405nm to 2.00abs 1) Turn the microplate around (A1 is now in the H12 position and repeat the test procedure described in the Repeatability section.
Page 135 This appendix contains the procedure for decontaminating the PowerWave. Decontamination Procedure If the PowerWave is to be shipped after being exposed to potentially hazardous material, it should be decontaminated. The following procedure outlines how to decontaminate the instrument before packaging and shipment.
Page 136: Appendix A: Decontamination
Procedure • A solution of .5% Sodium Hypo Chlorite (NaOCL) solution (Bleach) is used. Commercial bleach is 5% NaOCL; household bleach is 3% NaOCL. When using commercial bleach use a 10:1 mixture; if using 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.
Page 137: Appendix B: Computer Control
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’s KCJR, KCII, KC3 or KC4 for...
Page 138 PowerWave as the reader. COMPUTER CONTROL PROGRAMMING PowerWave’s RS-232C serial port 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.
Page 139 SIGN CHARACTERS Positive value / not designated as a blank well Negative value- / not designated as a blank well Out-of-range value-/ not designated as a blank well Positive value / used as a blank well Negative value- / used as a blank well Out-of-range value / used as a blank well...
Page 140 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: Halt the motors that move the micro plate and the filter wheel;...
Page 141 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 string to the computer. When the reader sends data to the computer, the data is sent first, then the status string.
Page 142 Computer Control Command Set A command from the computer to the reader consists of a single ASCII character, and, in some cases, a supporting argument. Table B-3 describes the supported computer control command set. Command Function Move the microplate carrier to its full-out position, then move inward into the reading chamber.
Page 143 Command Function Set wavelengths (see page B-19 for description). 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.) Appendix B...
Page 144 Number of Data bytes Checksum for this file structure. Loop through each byte of the following array, starting at 2. Put that number in Array 1. (This is the checksum for this structure.) NOT USED 2 - 7 8 - 29 NOT USED Options Bit Set to 1...
Page 145 Number of Data bytes Filter #2 (ASCII filter #) 53 - 55 Read mode (0 or 2) 0 Read-and eject 2 Repeated reads\Kinetics NOT USED 58-59 NOT USED Read timing value 11 - 9999 seconds between repeated reads on single 60-61 wavelength;...
Page 146 Number of Data bytes Bit 6: 1 sets this well as the blank well; 0 does not set this well as a blank well. Bit 7: Not used 163-165 166-167 Length of read in Bit Set to 1 Bit Set to 0 minutes for TTL minutes in Repeated-Reads...
Page 147 Status String Format Following execution of each command, PowerWave sends a status string back to the computer if Return of Status is enabled. This string consists of 5 ASCII characters -- RS, S3, S2, S1, and ETX: ⇒ RS A record separator that marks the beginning of the status string.
Page 148 Example of Application Subroutine XAMPLE Goal: Read a microplate at 405 nm, blank on well A1, print the heading and optical densities for all wells, and send the optical densities for all wells to the computer. • Set up a 170-character array (see Table B-4). Each character should be 1 byte.
Page 149 Array [53-55] Read and eject. Array [56] Array [57] = 0, 0 Array [58-59] low byte, high byte is the order = 0, 0 low byte, high byte is the order Array [60-61] = 0, 0 Array [62-63] low byte, high byte is the order Array [64] Full blank map.
Page 150 Note: Each of the 96 wells is given an 8-bit designation describing what sector it is in, if the sector is to be read, and if it is to be used for blanking. The first 4 digits (bits 0 - 3), describe the sector number, and in this case the sector is sector 1.
Page 151 Send this setup to the reader. Send each character out the serial port using the appropriate command from the asynchronous computing package used. Cmdstr = V(170byte array) Check for Status =000, which is an automatic return message from the reader. Start the reader.
Page 152 NOTE: All ASCII numeric representations send high digit first. INSTRUMENT IDENTIFICATION When a single '}' character is sent to the spectral reader, a PowerWave 200 will respond with a single ‘0’. A PowerWave 340 will response with a single ‘1’. All other instruments will return <NAK>. STRIP READS The Select Strips (‘%’) command is defined as follows:...
Page 153 If only a single strip is to be read, both bytes should be set to the same value, i.e. 0x03, 0x03 indicates read strip 3 only. If the last strip is lower than the first strip, only the first strip will be read. A first strip argument of 0x00 will cause the entire plate to be read (as will 0x01, 0x0C).
Page 154 MULTIPLE WAVELENGTH READS To cause a read to return data for each indicated wavelength (rather than performing dual wavelength subtracts or blanking), use the following procedure: a) Set both wavelengths in the standard reader definition table (see ‘V’ command) to '000'. In this case, the single/dual wavelength select parameter (byte #30, bit #6) will be ignored.
Page 155 An aborted read is indicated by a <DLE> response at any time. Any data received, including checksums, should be considered invalid and irrelevant in this case. After data for the very last plate has been sent, the reader performs some final FMEA tests to ensure that no errors may have occurred during the read process.
Page 156 Each wavelength is represented by 'xxx,'. (3 ASCII digits followed by a comma). Six wavelengths must be sent (24 bytes), followed by ETX after last comma. If a wavelength is not to be specified, it must be replaced by '000'. The Set Filter Table command returns the standard status response string.
Page 157 calibrate option (1) '0': calibrate only if necessary '1': calibrate before scanning shake time (2) '00' - '99': time in seconds shake speed (1) '0': slow '1': medium '2': fast '3': variable <ETX> <ACK/NAK> The standard status response string is then returned to the host. If an error code is returned with the status, it indicates that the spectral scan has terminated, and no further response data will be transmitted by the reader.
Page 158 For each wavelength: wavelength (3) '200'-'800' For each well selected (1 or 8): comma separator (1) ‘,’ sign (1) '+' or '-' data (4) ‘xxxx' well terminator <CR> <LF> data terminator <^Z> checksum (1) 0x00 - 0xFF If the calibrate option is selected, all wavelengths indicated for the current spectral scan will be calibrated.
Page 159 TEMPERATURE POLLING The Get Temperature (‘]‘) command is defined as follows: Host Response <ACK/NAK> 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.
Page 160 The Get Minimum Kinetic Interval (‘$‘) command is defined as follows: Host Response <ACK/NAK> Reader response protocol (hex format): interval low byte,0x00 0xFF interval high byte,0x00 0xFF The Get Minimum Kinetic Interval command then returns the standard status response string. Intervals are returned in units of seconds.
Page 161: Appendix C: Error Codes
A p p e n d i x C Error Codes This is a list of Error Codes that may appear on the reader. If an error is displayed, call Bio-Tek’s Technical Assistance Center between 8:30 AM and 6:00 PM Eastern Standard Time, Monday through Friday, except holidays.
Page 162 ROBABLE CAUSE • Carrier Axis Case - Bad sensor. The X-axis movement is limited so that the optical sensor cannot be interrupted. • Filter Wheel Case - Bad sensor. The filter wheel movement is limited so that the sensor cannot be interrupted. Or, the filter wheel is not installed.
Page 163 Displayed Error Potential Cause ERROR 0400 Carrier axis failed position verify ERROR 0401 Filter Wheel motor failed position verify ERROR 0402 Monochromator failed position verify ERROR 0403 Robot failed position verify Errors 400, 401, 402 and 403 indicate that an axis failed its Position Verify test.
Page 164 Displayed Error Potential Cause ERROR 0601 LAMBDA #1 Gain out of range ERROR 0602 LAMBDA #2 Gain out of range ERROR 0603 LAMBDA #3 Gain out of range ERROR 0604 LAMBDA #4 Gain out of range ERROR 0605 LAMBDA #5 Gain out of range ERROR 0606 LAMBDA #6 Gain out of range Errors 601-606 indicate that the gain for a specific wavelength is out of the...
Page 165 Displayed Error Potential Cause ERROR 0800 Reader failed offset test Error 800 indicates that the measurement electronics’ dark current offset is outside of acceptable limits at maximum gain. ROBABLE CAUSE • Ambient light leak or faulty printed circuit board. Displayed Error Potential Cause ERROR 0900 Read time dark value out of...
Page 166 ROBABLE CAUSE The measurement electronics full signal level has changed since the power-up self-check was last run. Displayed Error Potential Cause ERROR 0C00 Printer timeout error Error C00 indicates that the time allotted for the instrument to make a valid connection to a printer has expired. ROBABLE CAUSE Printer not connected or powered up.
Page 167 ERROR 1500 Incubator failed to hold temperature within tolerances during the assay. ERROR 1600 Computer 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.
Page 168 Error Codes...
Page 169: Appendix D: Report Format
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...
Page 170 Figure 1: Samples with Calls on Matrix Report Report Format...
Page 171 Figure 2: Curve Fit Report Appendix D...
Page 172 Figure 3: Samples with Calls on Column Report Report Format...
Page 173 Figure 4: Column Report without Samples Appendix D...
Page 174 Figure 5: Panel Report Report Format...
Page 175 Figure 6: Assay Detail Report Appendix D...
Page 176 Figure 7: Assay List Report Format...

Biotek PowerWave Operator's Manual

Warnings and Precautions

Intended Use Statement

Chapter 1 Introduction

Introducing the Powerwave

Chapter 2: Installation

The Operating Environment

Unpacking and Repackaging the Instrument

After Unpacking, Verify Performance

Before Repackaging the Instrument

Adjusting the Reader's Wavelength Table Setting

Printing and Data Communications

Lamp Replacement

Adjusting the Line Voltage Input Range

Adjusting the Fuse Configuration or Fuse Replacement

Other Utility Options

Chapter 3: Operation

Introduction

The Startup Screen

The Main Menu Screen

Define

Define (Method, Map, Formula and Curve)

Method

Map Definition

Formula Definition

Defining the Curve-Fit

Reading a Microplate

Chapter 4: Performance Verification

Achieving Maximum Performance

Installation and Operational Qualification Procedures

Routine Procedure

Accessing Tests and Calibration Using the Utility Option

Test Options

Calibration Validation

Running the Calibration Plate Test

Linearity Test

Verification of Alignment

Appendix A: Decontamination

Appendix B: Computer Control

Appendix C: Error Codes

Appendix D: Report Format

Quick Links

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Summary of Contents for Biotek PowerWave

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