multi-mode validation plate user guide -...

Multi-Mode Validation Plate

For the SpectraMax® Paradigm® Multi-Mode Detection Platform and the FilterMax™ Multi-Mode Microplate Readers

User Guide

5008531 ASeptember 2010

This document is provided to customers who have purchased Molecular Devices, Inc. (“Molecular Devices”) equipment, software, reagents, and consumables to use in the operation of such Molecular Devices equipment, software, reagents, and consumables. This document is copyright protected and any reproduction of this document, in whole or any part, is strictly prohibited, except as Molecular Devices may authorize in writing.

Software that may be described in this document is furnished under a license agreement. It is against the law to copy, modify, or distribute the software on any medium, except as specifically allowed in the license agreement. Furthermore, the license agreement may prohibit the software from being disassembled, reverse engineered, or decompiled for any purpose.

Portions of this document may make reference to other manufacturers and/or their products, which may contain parts whose names are registered as trademarks and/or function as trademarks of their respective owners. Any such usage is intended only to designate those manufacturers' products as supplied by Molecular Devices for incorporation into its equipment and does not imply any right and/or license to use or permit others to use such manufacturers' and/or their product names as trademarks.

Molecular Devices makes no warranties or representations as to the fitness of this equipment for any particular purpose and assumes no responsibility or contingent liability, including indirect or consequential damages, for any use to which the purchaser may put the equipment described herein, or for any adverse circumstances arising therefrom.

For research use only. Not for use in diagnostic procedures.

The trademarks mentioned herein are the property of Molecular Devices, Inc. or their respective owners. These trademarks may not be used in any type of promotion or advertising without the prior written permission of Molecular Devices, Inc.

Product manufactured by Molecular Devices, Inc.

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Molecular Devices, Inc. is ISO 9001 registered.

© 2010 Molecular Devices, Inc.

All rights reserved.

Printed in Austria.

Safety Information

All Warnings and Cautions in this document include an exclamation point, a lightning bolt, or a light burst symbol framed within a triangle. Please pay special attention to the specific safety information associated with these symbols.If the equipment is used in a manner not specified, the protection provided by the equipment may be impaired.

Warning and Caution DefinitionsThe exclamation point symbol is an international symbol which serves as a reminder that all safety instructions should be read and understood before installation, use, maintenance, and servicing is attempted.When this symbol is displayed in this guide, pay special attention to the specific safety information associated with the symbol.

CAUTION! A CAUTION calls attention to a condition or possible situation that could damage or destroy the product or the operator’s work.

Electrical SafetyTo prevent electrically related injuries and property damage, properly inspect all electrical equipment prior to use and immediately report any electrical deficiencies. Contact a service representative for any servicing of equipment requiring the removal of covers or panels.

WARNING! A WARNING calls attention to a condition or possible situation that could cause injury to the operator.

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Safety Information

High Voltage

Do not remove system covers. To avoid electrical shock, use supplied power cords only and connect to properly grounded (three-holed) wall outlets. Do not use multi-plug power strips.

Chemical and Biological SafetyNormal operation of the Multi-Mode Validation Plate may involve the use of materials that are toxic, flammable, or otherwise biologically harmful. When using such materials, observe the following precautions:

• Handle infectious samples according to good laboratory procedures and methods to prevent the spread of disease.

• Observe all cautionary information printed on the original solutions containers prior to their use.

• Dispose of all waste solutions according to your facility’s waste disposal procedures.

• Operate the Multi-Mode Validation Plate in accordance with the instructions outlined in this manual, and take all the necessary precautions when using pathological, toxic, or radioactive materials.

• Splashing of liquids may occur; therefore, take appropriate safety precautions, such as using safety glasses and wearing protective clothing, when working with potentially hazardous liquids.

• Use an appropriately contained environment when using hazardous materials.

WARNING! This symbol indicates the potential of an electrical shock hazard existing from a high voltage source and that all safety instructions should be read and understood before proceeding with the installation, maintenance, and servicing of all modules.

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Multi-Mode Validation Plate User Guide

• Observe the appropriate cautionary procedures as defined by your safety officer when using flammable solvents in or near a powered-up instrument.

• Observe the appropriate cautionary procedures as defined by your safety officer when using toxic, pathological, or radioactive materials.

Moving PartsTo avoid injury due to moving parts, observe the following:

• Never attempt to exchange labware, reagents, or tools while the instrument is operating.

• Never attempt to physically restrict any of the moving components of the instrument when using the Multi-Mode Validation Plate.

• Keep the instrument work area clear to prevent obstruction of the movement.

CleaningObserve the cleaning procedures outlined in this user guide for the Multi-Mode Validation Plate. Before cleaning equipment that has been exposed to hazardous material:

• Appropriate Chemical and Biological Safety personnel should be contacted.

• The Chemical and Biological Safety information contained in this user guide should be reviewed.

Note: Observe all warnings and cautions listed for any external devices attached or used during operation of the Multi-Mode Validation Plate. Refer to applicable external device user’s manuals for operating procedures of that device.

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Safety Information

MaintenancePerform only the maintenance described in this guide. Maintenance other than that specified in this guide should be performed only by service engineers.

Warnings and Cautions Found in this GuidePlease read and observe all cautions and instructions. Remember, the most important key to safety is to operate the Multi-Mode Validation Plate with care.The WARNINGs and CAUTIONs found within this document are listed below.

CAUTION! The validation plate must have the same hardware ID number as that selected in the protocol configuration and must be placed on the microplate carrier in landscape orientation with well A1 in the back-left corner. The validation read will fail if these conditions are not met.

Note: It is your responsibility to decontaminate components of the Multi-Mode Validation Plate before requesting service by a service engineer or returning parts to Molecular Devices for repair. Molecular Devices will NOT accept any items which have not been decontaminated where it is appropriate to do so. If any parts are returned, they must be enclosed in a sealed plastic bag stating that the contents are safe to handle and are not contaminated.

WARNING! If the equipment is used in a manner not specified, the protection provided by the equipment may be impaired.

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CAUTION! Handle the validation plate by the edges only. Contaminants on the standards, such as dust or fingerprints may cause tests to fail unexpectedly.

CAUTION! Clean standards with the supplied SpeckGRABBER tool only. Other cleaning tools, such as brushes, compressed air, solvents, or cotton swabs can damage the standards.

CAUTION! Never touch the SpeckGRABBER cleaning pad. Oil from fingerprints can be transferred to the surfaces of standards during cleaning. The cleaning pad should be cleaned before each use.

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Safety Information

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Contents

Safety Information. . . . . . . . . . . . . . . . . . . . . 3Warning and Caution Definitions . . . . . . . . . . 3Electrical Safety . . . . . . . . . . . . . . . . . . . . . 3

High Voltage . . . . . . . . . . . . . . . . . . . . . . 4Chemical and Biological Safety . . . . . . . . . . . 4Moving Parts . . . . . . . . . . . . . . . . . . . . . . . . 5Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . 5Maintenance . . . . . . . . . . . . . . . . . . . . . . . . 6Warnings and Cautions Found in this Guide . . 6

Chapter 1 Multi-Mode Validation Plate Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

Introduction . . . . . . . . . . . . . . . . . . . . . . . 11Absorbance Validation Overview . . . . . . . . 13Fluorescence Validation Overview . . . . . . . 16Luminescence Validation Overview. . . . . . . 21

Chapter 2 Installing the Multi-Mode Validation Plate Software . . . . . . . . . . . . . . 25

Introduction . . . . . . . . . . . . . . . . . . . . . . . 25Before Installing or Updating the Software. . . . . . . . . . . . . . . . . . . . . . . . . 26Installing the Multi-Mode Validation Plate Software on Windows XP . . . . . . . . . . . . . 27Installation Use Cases . . . . . . . . . . . . . . 30

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Contents

Chapter 3 Multi-Mode Validation Plate Tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Performing a Validation Read . . . . . . . . . . . 33Viewing the Results of a Validation Read . . . 38Printing or Exporting the Results of a Validation Run . . . . . . . . . . . . . . . . . . . 45

Chapter 4 Basic Maintenance and Troubleshooting. . . . . . . . . . . . . . . . . . . . . . 47

Overview . . . . . . . . . . . . . . . . . . . . . . . . . 47Cleaning the Multi-Mode Validation Plate . . . 47Replacing the Multi-Mode Validation Plate Batteries . . . . . . . . . . . . . . . . . . . . . . . . . 49Updating the Electronic Certificate After Recalibration. . . . . . . . . . . . . . . . . . . 51Troubleshooting the Multi-Mode Validation Plate and Test Result Failures . . . 53Handling Reproducible Failures . . . . . . . . . 55

Appendix A Available Validation Tests . . . . 57Tests and Supported Hardware . . . . . . . . . . 57Tests with Parameters . . . . . . . . . . . . . . . . 61

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1
Multi-Mode Validation Plate Overview
IntroductionThe Multi-Mode Validation Plate (Figure 1-1) is used to validate the performance of absorbance, fluorescence, and luminescence measurements performed on the following Molecular Devices instruments:

• SpectraMax® Paradigm® Multi-Mode Detection Platform

• FilterMax™ F3 Multi-Mode Microplate Reader• FilterMax™ F5 Multi-Mode Microplate Reader

Validation tests performed with the validation plate only validate detector performance. Factory-defined and user-defined settings are not changed.Multi-Mode Validation Plate validation tests differ for the SpectraMax Paradigm Multi-Mode Detection Platform, FilterMax F5 Multi-Mode Microplate Reader, and FilterMax F3 Multi-Mode Microplate Reader. Refer to Tests and Supported Hardware on page 57 for the instruments supported by each test. Refer to Tests with Parameters on page 61 for an overview of available tests. For more information about various types of instrument measurement tests, refer to the appropriate section of this overview:

• Absorbance Validation Overview on page 13• Fluorescence Validation Overview on page 16• Luminescence Validation Overview on page 21

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Validation tests are performed in the Multi-Mode Analysis Software using a special validation protocol. Results from validation reads are saved in a software database and can be accessed from the Results Selection List at any time.

This guide assumes the user has a good working knowledge of the Multi-Mode Analysis Software and covers topics specific to the validation plate only. Topics covered include:

• Multi-Mode Validation Plate Overview on page 11

Figure 1-1 Multi-Mode Validation Plate

Item Description

1 Well A1

2 Validation plate luminescence On button

3 Battery compartment cover

4 Hardware ID number

901840L.AI

1

3

2

4

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• Installing the Multi-Mode Validation Plate Software on Windows XP on page 27

• Multi-Mode Validation Plate Tests on page 33 • Basic Maintenance and Troubleshooting on

page 47

Absorbance Validation OverviewAbsorbance parameters are tested using the standards shown in Figure 1-2. Table 1-1 on page 14 provides descriptions of the parameters validated in absorbance tests.

Figure 1-2 Absorbance Standards

Item Description

1 Absorbance X alignment check

2 Absorbance functional checks

3 Absorbance Y alignment check

4 Absorbance standard curve

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Table1-1 Absorbance Parameters validated by the Multi-Mode Validation Plate

Parameter Details

Functionality • Beam Intensity: Validates that the absorbance beam is within the operating range of the detector.

• Filter Integrity: Validates the absorbance filter’s proper out-of-band rejection.

• Wavelength Plausibility (FilterMax Multi-Mode Microplate Readers only): Validates the absorbance filter slide configuration to ensure filter definitions are assigned to the correct slots.

• High Low: Validates the extreme high and low limits of the measurement range.

Alignment Quantifies the alignment between the Multi-Mode Validation Plate and the absorbance read head of the detector. Measured values are compared to values calibrated at the factory, which represent the proper alignment of a Society of Biomolecular Screening (SBS) standard plate.

Note: Use the Multi-Mode Analysis Software labware optimization feature to compensate for dimensional variations between production lots of microplates.

Precision Validates repeatability between kinetic cycles. Precision is reported as the standard deviation of the repeated readings.

Note: The Multi-Mode Validation Plate hardware features a standard curve covering the absorbance measurement range for the standard wavelengths supported by the instrument. This feature is required to determine precision for the entire measurement range.

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Linearity Validates the linearity of optical density (OD) measurements by comparing the measured values to NIST-traceable standard values.

Note: The Multi-Mode Validation Plate hardware features a standard curve covering the absorbance measurement range for the standard wavelengths supported by the instrument. This feature is required to determine linearity.

Accuracy Validates the reading accuracy of OD measurements by comparing the measured values to NIST-traceable standard values.

Wavelength Accuracy

SpectraMax Paradigm Multi-Mode Detection Platform only: Validates the wavelength accuracy of OD measurements by comparing the measured values to NIST-traceable standard values.

Table1-1 Absorbance Parameters validated by the Multi-Mode Validation Plate (cont’d)

Parameter Details

Note: The printed certificate included with the Multi-Mode Validation Plate provides the standard curve of NIST-traceable absorbance values (in absorbance units) as determined on equipment regularly calibrated using NIST-traceable absorbance standards.

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Fluorescence Validation OverviewFluorescence parameters are tested using the standards shown in Figure 1-3. Table 1-2 on page 17 provides descriptions of parameters validated in fluorescence tests.

Figure 1-3 Fluorescence Standards

Item Description

1 Fluorescence X alignment check

2 Fluorescence standard curve

3 Fluorescence functional checks and extended testing

4 Fluorescence Y alignment check

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Table1-2 Fluorescence Parameters Validated by the Multi-Mode Validation Plate

Parameter Details

Functionality • SpectraMax Paradigm Multi-Mode Detection Platform: Cartridges that cannot be supported by the validation plate hardware are tested for their functionality.

• Blocking (FilterMaxF5 Multi-Mode Microplate Reader only)*: The Blocking parameter validates the integrity of polarizing filters installed on FilterMax F5 Multi-Mode Microplate Reader filter slides.* Does not apply to FilterMax F3

Multi-Mode Microplate Reader.

Alignment Quantifies the alignment between the validation plate and the fluorescence read head of the detector. Measured values are compared to values calibrated at the factory, which represent the proper alignment of a Society of Biomolecular Screening (SBS) standard plate.


Measurement range

The validation plate hardware features a standard curve for the fluorescence top wavelengths. For details, refer to Available Validation Tests on page 57 (Appendix A). This feature is required to determine linearity.

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Precision Validates the precision of fluorescence top measurements by determining the kinetic cycle-to-cycle repeatability for each standard of the curve. For details, refer to Available Validation Tests on page 57 (Appendix A). Precision is reported as a coefficient of variation (CV) value.• SpectraMax Paradigm Multi-Mode Detection

Platform and FilterMaxF5 Multi-Mode Microplate Reader*: Also validates the precision of fluorescence polarization measurements.* Does not apply to FilterMax F3

Multi-Mode Microplate Reader.

Note: The Multi-Mode Validation Plate hardware features a standard curve covering the fluorescence measurement range for the standard wavelengths supported by the instrument. This feature is required to determine precision for the entire measurement range.

Linearity Compares values measured in the test with linear calibration values determined at the factory.

Note: The Multi-Mode Validation Plate hardware features a standard curve for the fluorescence top wavelengths. This feature is required to determine linearity.

Table1-2 Fluorescence Parameters Validated by the Multi-Mode Validation Plate (cont’d)

Parameter Details

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Signal • Determines the signal to background (S/B) ratio for a type of standard measured with a particular filter set. Using the equivalent label concentration of the standard (see the printed certificate), the reciprocal S/B is transformed into background equivalent label concentration. A small reciprocal S/B value indicates the proper combination of label, filter set, and light source is present.

• Determines the signal to background noise ratio (S/N) for a type of standard measured with a particular filter set. Using the equivalent label concentration of the standard (see printed certificate), the reciprocal S/N is scaled to the detectable equivalent label concentration. A small reciprocal S/N value indicates the baseline of the instrument is stable.

Note: Noise (N) is measured as the precision value of the blank standard on the Multi-Mode Validation Plate. In the assay used to determine instrument specifications, the detection limit is based on replicate uniformity of assay blanks. Passing the Validation Plate Signal test provides supporting evidence, but does not guarantee, that the instrument can achieve the published detection limits.


Parameter Details

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Dynamic range Quantifies the useful measurement range between the noise level N (three times the standard deviation of repeated blank reads) and signal S at the higher end of the standard curve of the Multi-Mode Validation Plate. Dynamic range equals LOG10(S/N).

Note: When parameter Linearity passes (indicated as PASS in the validation plate results), the Dynamic Range may be regarded as the Linear Dynamic Range.


Parameter Details

Note: The printed certificate included with the Multi-Mode Validation Plate provides the standard curve of fluorescein in units of an equivalent fluorescein concentration measured under standard conditions using NIST SRM 1932 (available through Invitrogen.com, article number F36915).

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Luminescence Validation OverviewLuminescence parameters are tested using the powered luminescence standards on row H of the validation plate (see Figure 1-4). Table 1-3 on page 21 provides descriptions of the parameters validated in luminescence tests.

Figure 1-4 Luminescence Standards

Item Description

1 Validation plate luminescence On button

2 Luminescence standard curve (row H)

Table1-3 Luminescence Parameters Validated by the Multi-Mode Validation Plate

Parameters Description

Functionality SpectraMax Paradigm Multi-Mode Detection Platform: Cartridges that cannot be supported by the validation plate hardware are tested for their functionality. For details, refer to Available Validation Tests on page 57.

901843L.AI

1

2

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Alignment Quantifies the alignment between the Multi-Mode Validation Plate and the luminescence read head of the instrument. Measured values are compared to values calibrated at the factory, which represent the proper alignment of an SBS standard plate.


Precision Validates the precision of luminescence measurements by determining the kinetic cycle-to-cycle repeatability for each standard. Precision is reported as a CV value.

Note: The validation plate hardware features a standard curve covering the luminescence measurement range for the standard wavelengths supported by the instrument. This feature is required to determine precision for the entire measurement range.

Linearity Compares values measured in the test with linear calibration values determined at the factory.

Note: The Multi-Mode Validation Plate hardware features a standard curve covering the luminescence measurement range for the standard wavelengths supported by the instrument. This feature is required to determine linearity.

Table1-3 Luminescence Parameters Validated by the Multi-Mode Validation Plate (cont’d)


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Signal • Determines the signal to background ratio (S/B) for the luminescence standard. Using the equivalent label concentration of the standard (see the printed certificate), the reciprocal S/B is transformed into background equivalent label concentration. The reciprocal S/B value (smaller values are better) indicates proper light collection and detection efficiency, which are prerequisites to achieving the detection limit of the instrument.

• Determines the signal to background noise ratio (S/N) for the luminescence standard. Using the equivalent label concentration of the standard (see printed certificate), the reciprocal S/N is transformed into the detectable equivalent label concentration.The reciprocal S/N value (the smaller the better) indicates the baseline of the instrument is properly stable, which is a prerequisite to achieve the detection limit of the instrument.

Note: Noise (N) is measured as the precision value of the blank standard on the Multi-Mode Validation Plate. In the assay used to determine instrument specifications, the detection limit is based on replicate uniformity of assay blanks. Passing the Validation Plate Signal test provides supporting evidence, but does not guarantee, that the instrument can achieve the published detection limits.

The PerkinElmer ATPLiteTM glow assay was used to determine the detection limit specification of the instrument.



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Dynamic range

Quantifies the useful measurement range between the noise level N (three times the standard deviation of repeated blank reads) and signal S at the higher end of the standard curve of the Multi-Mode Validation Plate. Dynamic range equals LOG10(S/N).

Note: When parameter Linearity passes (indicated as PASS in the validation plate results), the Dynamic Range may be regarded as the Linear Dynamic Range.



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2
Installing the Multi-Mode Validation Plate Software
IntroductionThe Multi-Mode Validation Plate Software installation CD contains components required to:

• install or update the Multi-Mode Validation Plate Software.

• install or update the Multi-Mode Validation Plate Electronic Certificate.

When you intend to work with an already installed software, and just update your certificate, you might proceed to section Updating the Electronic Certificate After Recalibration on page 51.

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Before Installing or Updating the SoftwareBefore you begin to install or update your Multi-Mode Validation Plate Software, please consider the following:The Multi-Mode Validation Plate requires that the Multi-Mode Analysis Software is already installed on your Multi-mode System Controller PC. To prevent unintentional update of your instrument, please check the following statements, before proceeding to follow the step-by-step instructions.

• When operating the instrument in an integrated system, contact your local Technical Support Representative to check the compatibility of the integration with the Multi-Mode Analysis Software version required for the Multi-Mode Validation Plate.

• When operating the instrument with validated protocols, contact your local Laboratory Manager to check, whether an update of the instrument complies with your laboratory regulations.

For the optimum support of your instrument, we generally recommend that you update to the most recent version. If you have accidentally updated following the next steps, and now need to downgrade, please contact your local Technical Support Representative for assistance.

Note: When confirming to proceed, please follow the step by step instructions in the following section. Remember the five digit HW ID number (hardware identifier: 5xxxx) on your Multi-Mode Validation Plate. This number can be found on the label on top of the Multi-Mode Validation Plate Toolbox, or on the printed certificate. It is also engraved on top of the Multi-Mode Validation Plate.

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Installing the Multi-Mode Validation Plate Software on Windows XPThe Multi-Mode Validation Plate Software installer uses a simple interface to guide the installation of required components and the Multi-Mode Validation Plate Software.

CAUTION! Before installing or updating components, please read Before Installing or Updating the Software on page 26.

To check versions and compatibility of the required components:

1. Exit all Windows programs before running the installer program.

2. Make sure the current user account has Administrator privileges. Accounts with Standard or Restricted access are not permitted to run the installer program. Contact the site system administrator for more information about account privileges.

3. Insert the Multi-Mode Validation Plate Software CD into the CD drive and browse to the contents of the CD.

4. Double-click Installer.exe.

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The Multi-Mode Validation Plate Software Installer displays all currently installed components (Figure 2-1): Multi-Mode Analysis Software (by version) is a

prerequisite to install or update the Multi-Mode Validation Plate Framework and Application components.

Multi-Mode Validation Plate Framework (by version) is a part of the Multi-Mode Validation Plate Software installation disks. It integrates the Multi-Mode Validation Plate Application with the Multi-Mode Analysis Software.

Multi-Mode Validation Plate Application is a part of the Multi-Mode Validation Plate Software installation disks and is the Multi-Mode Validation Plate Software Plug In to the Multi-Mode Analysis Software.

Available Plates is a list of electronic certificates available for install or update. Running the Multi-Mode Validation Plate requires the one certificate that matches the hardware ID of your Multi-Mode Validation Plate (compare item 4 in Figure 1-1 on page 12).

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Figure 2-1 Starting the Software Installation

Item Description

1 Certificates for plate 50139 and 50142 installed

2 Certificate update for plate 50140 on the CD

3 V1: compatible with Validation Plate Application < 3.2

4 V3: compatible with Validation Plate Application > 3.1

1

2

43

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Installation Use Cases

Use Case A: Multi-Mode Analysis Software • Multi-Mode Analysis Software not installed: Use the

Multi-Mode Analysis Software Installation disks to install the required basis for the Multi-Mode Validation Plate Software. Please read Before Installing or Updating the Software on page 26.

• Multi-Mode Analysis Software installed, but version is not current: When an older version of the Multi-Mode

Validation Plate Software is also installed: You can continue using the already installed software, and install or update one or more plates under Use Case D: Available Plates on page 32. An update of the certificate is required after recalibration of the plate at factory.

First installation of the Multi-Mode Validation Plate Software: You might need to update the Multi-Mode Analysis Software to the indicated version. Use the Multi-Mode System Updater Package Installation disks. Please read Before Installing or Updating the Software on page 26.

Note: When a component installation requires restarting the system, restart before installing the next component listed. Failure to restart can result in unwanted software actions. After the system restarts, browse to the contents of the installer CD and relaunch the installer to continue installing components.

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Use Case B: Multi-Mode Validation Plate Framework • Former version of Multi-Mode Validation Plate

Software is installed: You can continue working with the already installed Software and proceed to install or update one or more plates under Use Case D: Available Plates on page 32.

• Installation or Update of Multi-Mode Validation Plate Software: Click the Install/Update button in this section of the installer window to start the installation of the Multi-Mode Validation Plate Software.

Use Case C: Multi-Mode Validation Plate Application • Former version of Multi-Mode Validation Plate

Software is installed: You can continue working with the existing Software and proceed to Use Case D: Available Plates on page 32.

• Installation or Update of Multi-Mode Validation Plate Software: Click the Install/Update button in this section of the installer window to continue the installation of the Multi-Mode Validation Plate Software.

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Use Case D: Available Plates • The Multi-Mode Validation Plate was already installed:

The Installer automatically detects the Hardware ID of your previously installed Multi-Mode Validation Plate. If there is a more current certificate for your plate on the installation disks, you will be prompted to update your certificate that might be expired, even before the installer window Figure 2-1 appears. For details, see Updating the Electronic Certificate After Recalibration on page 51.

• First Install of a Multi-Mode Validation Plate: Scroll through the available certificates and select the HW ID number of your Multi-Mode Validation Plate by clicking in the list. Select suffix -V1 for use with FilterMax

Multi-Mode Microplate Readers and Software version 3.1 or lower.

Select suffix -V3 when working with Software Version 3.2 or higher. V3 is compatible with both the SpectraMax Paradigm Multi-Mode Detection Platform and the FilterMax Multi-Mode Microplate Readers.

Click Install to install the selected certificate. In the Multi-Mode Validation Plate Software

Installer, click the Close button. The Multi-Mode Validation Plate can now be used with the software. See Performing a Validation Read on page 33.

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3
Multi-Mode Validation Plate Tests
Performing a Validation ReadValidation reads are performed using the Multi-Mode Validation Plate Validation protocol.To perform a validation read:

1. Click the Start > Molecular Devices > Multi-Mode Analysis Software > Multi-Mode Analysis Software. The Multi-Mode Analysis Software opens.

2. In the Protocol Selection List pane, click the Validation Plate protocol.

Figure 3-1 Selecting Multi-Mode Validation Plate Test Protocol

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3. From the toolbar, click Run button. The Validation Plate Protocol Run window appears.

Figure 3-2 Selecting Tests to Perform in a Validation Run on a FilterMax™ F5 Multi-Mode Microplate Reader

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Figure 3-3 Selecting Tests to Perform in a Validation Run on the SpectraMax Paradigm Multi-Mode Detection Platform

4. In the Select a plate field, select the hardware ID number of the Multi-Mode Validation Plate being used to perform the validation. The hardware ID number is engraved on the Multi-Mode Validation Plate.

5. Select the tests desired to perform. Each test lists the excitation and/or emission filter slides used in the test and the time required to complete the test.

6. Click Next to start the validation read. The microplate carrier extends from the detector.

CAUTION! The Multi-Mode Validation Plate must have the same hardware ID number as that selected in the protocol configuration and must be placed on the microplate carrier in landscape orientation with well A1 in the back left corner. The validation read may fail if these conditions are not met.

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CAUTION! Handle the Multi-Mode Validation Plate by the edges only. Contaminants on the standards, such as dust or fingerprints can cause tests to fail unexpectedly.

CAUTION! When not in use, return the Multi-Mode Validation Plate to its toolbox. Keep the toolbox closed to prevent contamination by dust and aerosols.

7. Place the validation plate that has the hardware ID number entered in Step 4 on the microplate carrier in landscape orientation with well A1 in the back-left corner.

8. Place the Multi-Mode Validation Plate into the detector and click OK. The Run Selected Tests window appears (see Figure 3-4) and the validation read starts.

Note: If the luminescence test is selected, an instruction dialog prompts to turn the Multi-Mode Validation Plate on. Press the Multi-Mode Validation Plate luminescence ON button. The green LED illuminates for approximately two seconds to indicate that the Multi-Mode Validation Plate is powered on.

If the green LED does not illuminate when the Multi-Mode Validation Plate is turned on, see Troubleshooting the Multi-Mode Validation Plate and Test Result Failures on page 53.

Note: Selected tests are performed in the order listed in the Run Selected Tests window. Depending on the tests selected, the detector might appear to be idle during the validation run. These delays are normal. Table 3-1 describes common delays that occur during validation runs.

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Figure 3-4 Validation Run in Progress

Note: FilterMax Multi-Mode Microplate Reader tests require the standard filter slides in factory configuration. If the required testing filter slide is not currently inserted in the detector, the test will not run. A message will display indicating which slide must be inserted.

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Viewing the Results of a Validation ReadThe results of a validation read are displayed in the Result Viewer window. Results are displayed in three screens:

• Test Summary displays information about the validation read and the Multi-Mode Validation Plate, and it reports results for each test performed.

• Test Detail displays the results reported by all parameters measured in the tests performed.

• Test Data displays the actual data for each precision test parameter measured. For example, luminescence, absorbance, and fluorescence top standard curves. The acceptable upper and lower limits for each test parameter are also displayed where applicable.

Results can also be printed or exported to a text file (see Printing or Exporting the Results of a Validation Run on page 45).

Table3-1 Delays That Can Occur During Validation Runs

Test Type Delay

Absorbance <= 340nm (FilterMax F5 Multi-Mode Microplate Reader only)

A 30-second delay occurs to allow the deuterium lamp to warm up before performing this test.

Fluorescence(SpectraMax Paradigm Multi-Mode Detection Platform and FilterMax F5 Multi-Mode Microplate Reader only)

The detector performs a read height optimization before performing fluorescence tests, resulting in a one minute delay.

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To view the results of a validation read:• If validation read has just completed: The Result

Viewer appears, including the Test Summary (Figure 3-5).

• If you are viewing a previous validation read: In the Results Selection List at left, select the desired results and, from the toolbar, click View the selected result. The Test Summary appears (Figure 3-5).

Figure 3-5 Viewing the FilterMax Multi-Mode Microplate Reader Test Summary

Note: Validation read results can be located by using the Result Viewer search features to find results for all protocols named Multi-Mode Validation Plate.

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Figure 3-6 Viewing the SpectraMax Paradigm Multi-Mode Detection Platform Test Summary

1. In the Test Summary, click Next to view the results reported for all parameters tested. The Test Detail window appears (Figure 3-7).

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Figure 3-7 Viewing the FilterMax Multi-Mode Microplate Reader Results Reported by Each Parameter Tested

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Figure 3-8 Viewing the SpectraMax Paradigm Multi-Mode Detection Platform Results Reported by Each Parameter Tested

2. Click Next to view the actual data measured for each precision parameter tested. The Test Data window displays (Figure 3-9).

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Figure 3-9 Viewing the Actual FilterMax Multi-Mode Microplate Reader Data For Each Precision Parameter Measure

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Figure 3-10 Viewing the Actual SpectraMax Paradigm Multi-Mode Detection Platform Data for Each Precision Parameter Measured

3. Click Cancel or Close to close the Result Viewer.You can print or export the results, if desired. See Printing or Exporting the Results of a Validation Run on page 45.

Note: Results may be printed or exported from any of the results screens.

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Printing or Exporting the Results of a Validation RunThe results of the validation run displayed in the Result Viewer can be printed or exported to a text file.To print or export results:

1. From the toolbar, select the desired action. Table 3-2 describes the available actions.

Table3-2 Result Viewer Print and Export Actions

Button Action Description

Print results

Prints the results in a report formatted using the settings configured in the Multi-Mode Analysis Software.

Export results

Exports the results to a text file. Select the storage directory and file name for the exported results.Exported results contain the data displayed in the Results Viewer, as well as the raw data measured and results from tests that are not displayed in the Result Viewer. This information can be useful for troubleshooting unexpected test failures. See Troubleshooting the Multi-Mode Validation Plate and Test Result Failures on page 53.

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4
Basic Maintenance and Troubleshooting
OverviewUsers can perform several basic maintenance procedures to ensure the performance of the validation plate.

• Cleaning the Multi-Mode Validation Plate on page 47.

• Replacing the Multi-Mode Validation Plate Batteries on page 49.

• Updating the Electronic Certificate After Recalibration on page 51.

• Troubleshooting the Multi-Mode Validation Plate and Test Result Failures on page 53.

Cleaning the Multi-Mode Validation PlateUnder normal use, the design of the validation plate protects the standards from becoming easily contaminated. Most contamination can be avoided by storing the validation plate in the supplied case when not in use. Cleaning should be considered only when a test fails repeatedly during a validation read.Dirt, dust, or lint on the surface of a standard may cause a test to fail. When a test fails unexpectedly, examine the standards for contamination. Small particles of dirt, dust, or lint can be safely removed using the SpeckGRABBER tool provided with the check plate.Never use solvents or attempt to open the validation plate casing during cleaning. If contamination serious enough to warrant the use of a solvent is present, return the validation plate to your Molecular Devices Service Representative.

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To clean standards with the SpeckGRABBER tool:

CAUTION! Clean standards with the supplied SpeckGRABBER tool only. Other cleaning tools, such as brushes, compressed air, solvents, or cotton swabs can damage the standards.

1. Place the validation plate into the supplied case on a stable work surface to protect it from being dropped during cleaning.If cleaning must be performed outside the supplied case, place a clean, lint-free, folded towel on the work surface to protect the validation plate in case it is dropped.

2. Examine the standards for contamination. Individual dirt, dust, or lint particles can be removed safely with the SpeckGRABBER tool. If more serious contamination is present, return the validation plate to the manufacturer.

CAUTION! Never touch the SpeckGRABBER cleaning pad. Oil from fingerprints may be transferred to the surfaces of standards during cleaning. The cleaning pad should be cleaned before each use.

3. If necessary, clean the SpeckGRABBER cleaning pad. Detailed instructions are available at the manufacturer website: http://www.kinetronics.com.

Note: This section provides an overview of cleaning standards with the SpeckGRABBER tool. Complete instructions for the tool are available at the manufacturer website: http://www.kinetronics.com.

Note: Consider cleaning standards only if a test has failed unexpectedly.

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http://www.kinetronics.com


4. Make sure the cleaning pad is thoroughly dry before using the tool to remove contamination from standards.

5. Gently apply the cleaning pad to the surface of a standard where contamination is present. The high adhesion surface of the pad allows contamination to be removed without applying pressure or brushing the surface of the standard.

6. If the SpeckGRABBER tool is unable to remove the contaminant, return the validation plate to the manufacturer for service.

7. Rinse the cleaning pad with distilled water to remove contaminants removed from standards.

8. After allowing the cleaning pad to dry, return the SpeckGRABBER tool to the validation plate case.

Replacing the Multi-Mode Validation Plate BatteriesThe luminescence standards are powered by two 3 V lithium coin cell batteries (type CR2330). Battery life is approximately 250 validation reads, or one year of normal use. The batteries must be replaced when the green indicator LED on the plate does not light after depressing the validation plate ON button.Replacing the batteries requires a 1.5 mm Allen wrench, which is included in the toolkit supplied with the detector.To replace the batteries:

1. Leave the validation plate in the supplied case to protect the bottom of the plate from dust and other contaminants.

2. Use a 1.5 mm Allen wrench to remove the two screws that secure the battery compartment cover in place (see Figure 4-1).

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3. Remove the battery compartment cover and batteries.

4. Properly dispose of the lithium batteries according to applicable ordinances and regulations.

5. Place the new batteries in the battery compartment, with the positive (+) contact side facing the bottom of the plate.

Figure 4-1 Replacing the Multi-Mode Validation Plate Batteries

Item Description

1 1.5 mm Allen screws

2 Battery compartment cover

3 Type CR2330 3 V lithium coin cell batteries with the positive (+) contacts facing the bottom of the plate

901844L.AI

1

2

3

Note: Use only type CR2330 3 V lithium coin cell batteries for replacement.

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6. Replace the battery compartment cover and tighten the screws with the 1.5 mm Allen wrench.

7. Turn the validation plate on. The green LED lights, indicating the batteries were successfully replaced.

Updating the Electronic Certificate After Recalibration

The electronic certificate provided with the plate expires one year after validation plate issue or calibration. The Multi-Mode Validation Plate Software provides notification automatically when a certificate expires. Return the plate to Molecular Devices for recalibration when the certificate expires. A new electronic certificate is provided with the validation plate.To update the electronic certificate, please follow the instructions below:

1. Insert the Multi-Mode Validation Plate Software CD with the new electronic certificate into the CD drive and browse to the contents of the CD.

2. In the contents of the CD, double-click Installer.exe. The installer checks if a certificate is installed on the computer and if a new certificate is present on the CD. If both certificates are found, a message appears.If the installer does not locate either certificate, the Installer dialog appears (Figure 4-2). Proceed with instructions given in “First Install of a Multi-Mode Validation Plate” under Use Case D: Available Plates on page 32.

Note: If the LED does not light, check both batteries to make sure the positive (+) contact sides face the bottom of the validation plate.

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3. In the message, click Yes to update the certificate. The new certificate replaces the existing version, which is automatically archived. A new message appears, asking if additional certificates will be installed or updated If you click No to cancel updating the certificate. The Installer appears (Figure 4-2).

4. In the message dialog, click Yes to install or update another electronic certificate. The Installer appears.Click No to close the installer without installing the new certificate.

Figure 4-2 Installing an Electronic Certificate

5. To install another certificate, see Use Case D: Available Plates on page 32.

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Troubleshooting the Multi-Mode Validation Plate and Test Result Failures

Perform the following hardware and software troubleshooting techniques when necessary

Table4-1 Troubleshooting the Multi-Mode Validation Plate

If Then

The green LED does not light when the luminescence on button is pressed.

• Try pressing the button with the edge of a fingernail rather than a finger tip.

• While in a dark room, check for the visibility of a faint green glow in well H10

• Replace batteries (see Replacing the Multi-Mode Validation Plate Batteries on page 49).

• Ensure batteries are installed with the positive (+) side facing toward the bottom of the plate.

• Ensure there are no obstructions or lint between the contacts.

Absorbance, luminescence, or fluorescence test fails.

• Confirm that the validation plate is placed on the microplate carrier in landscape orientation with well A1 in the back-left corner.

• Retry the tests that fail to determine if the failure is reproducible. If the failure is reproducible, follow the steps in Handling Reproducible Failures on page 55.

Luminescence test fails.

• The validation plate was not powered on prior to performing the validation read. Press the validation plate luminescence ON button and repeat the test.

• The powered luminescence standards may have timed out before completing the test. Press the validation plate ON button and repeat the test.

• Retry the test to determine if the failure is reproducible. If the failure is reproducible, follow the steps in Handling Reproducible Failures on page 55.

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An absorbance test fails

• Visually inspect the standards in wells C3 to C5 in the third row and wells E3 to E10 in the fifth row for contaminants, such as dust, or damage, such as loose glass, cracks, or scratches.

• Remove minor contaminants such as dust, dirt, or lint particles with the provided SpeckGRABBER tool (see Cleaning the Multi-Mode Validation Plate on page 47).


A luminescence or fluorescence precision test fails.

• An emission filter slide must be present in the detector to power on the Photo Multiplier Tube (PMT).

Note: Allow the detector and PMT to warm up for at least thirty minutes before performing a validation read.


A fluorescence signal test fails

• Confirm that the filter sliders are in the standard factory configuration.

• Contaminants in row 6, well F2 can cause high background noise or invalidate the linearity test. The lower standard might read unexpectedly low or even negative. Inspect well F2 and clean with the provided SpeckGRABBER tool if dust, dirt, or lint is present (see Cleaning the Multi-Mode Validation Plate on page 47).



If Then

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Handling Reproducible FailuresReproducible failures indicate that the validation plate might require service. Before contacting Molecular Devices Technical Support, follow the steps in this section to help Technical Support Representatives determine the appropriate course of action.When a failure is reproducible:

1. Perform the entire set of tests available in the protocol. Determining the reason for a test failure is easier when it is also known which read modes and tests pass. Export the test results as described in the following steps and relay them to your Molecular Devices Technical Support Representative.

2. In the Result Viewer, click Export entire test record to a text file. The Save as dialog appears.

3. In the Save as dialog, browse to the desired directory where the exported file will be saved, provide a name for the file, and click Save. The complete results of the validation read are exported to a text file.

An alignment test fails.

• Perform an alignment test for a different read mode. If this alignment test passes, follow the steps in Handling Reproducible Failures on page 55.



If Then

Note: Result viewer printouts might not provide sufficient or specific information contributing to a test failure.

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4. If available, open the results from a previous validation read where all tests passed, and export these results to a text file by following Step 2 and Step 3.

5. Send your Molecular Devices Technical Support Representative an email containing: the exported text file of the results from the

validation read that failed. the exported text file of the results from a

previous validation read that passed (if available).

the date and details of the last service procedure performed on the reader.

any additional observations or concerns regarding the reader.

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A
Available Validation Tests
The following tables provide further detail on available validation plate tests, the parameters specific to these tests, and hardware supported by these tests.

Tests and Supported Hardware

TableA-1 Fluorescence Intensity Top Test

Label of Type

Wave-length

FilterMax F3 Reader

FilterMax F5 Reader

SpectraMax Paradigm Cartridge

Fluorescein 485/535 X X MULTI (0200-7001)FI-FLRH (0200-7003)

Rhodamine 535/595 X MULTI (0200-7001)FI-CY3CY5 (0200-7013)

Coumarin 360/465 X MULTI (0200-7001)FI-COFL (0200-7002)

Coumarin B 406/465 GENEBLAZER (0200-0714)MULTI-TOX (0200-7015)

Texas Red 585/635 MULTI (0700-7001)

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TableA-2 Fluorescence Intensity Bottom Test

Label of Type

Wave-length

FilterMax F3 Reader

FilterMax F5 Reader


Fluorescein 485/535 X MULTI (0200-7001)FI-FLRH (0200-7003)

Rhodamine 535/595 MULTI (0200-7001)FI-CY3CY5 (0200-7013)

TableA-3 Fluorescence Polarization Test

Label of Type

Wave-length

FilterMax F3 Reader

FilterMax F5 Reader


Fluorescein 485/535 X FP-FLUO (0200-7005)

Rhodamine 535/595 FP-RHOD (0200-7006)

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TableA-4 Fluorescence Time Resolved Test

Label of Type

Wave-length

FilterMax F3 Reader

FilterMax F5 Reader


Europium Eu/Eu X MULTI (0200-7001)TRF-EUSA (0200-7004)HTRF (0200-7007)

Eu-acceptor Eu/665 HTRF (0200-7007)

TableA-5 Luminescence Test

Label of Type

Wave-length

FilterMax F3 Reader

FilterMax F5 Reader


generic no filter X X MULTI (0200-7001)LUM (0200-7008)

TableA-6 Absorbance Test

Label of Type

Wave-length

FilterMax F3 Reader

FilterMax F5 Reader


generic fixed X X ABS (0200-7000)

genomic fixed X

generic scan ABS (0200-7000)

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TableA-7 Functionality Test

Label of Type

Wave-length

FilterMax F3 Reader

FilterMax F5 Reader


NA NA LUM-BRET2 (0200-7012)LUM-CHROM (0200-7009)LUM96 (0200-7010)LUM384 (0200-7011)ALPHA384 STD (0200-7016)ALPHA384 HTS (0200-7017)ALPHA 1536 HTS (0200-7018)FI-CFP-YFP (0200-7019)

Note: FilterMax™ Multi-Mode Microplate Reader tests require the standard filter slides in factory configuration.

Note: FilterMax F3 Multi-Mode Microplate Reader absorbance and fluorescence tests require excitation slide 3. Luminescence tests require emission slide 3 in factory configuration, in particular, position 6 must be empty.

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Tests with Parameters

Note: FilterMax F5 Multi-Mode Microplate Reader absorbance tests require excitation slide 2. Luminescence tests require emission slide 1 in factory configuration, in particular, position 6 must be empty. Fluorescence tests require excitation slide 1 and emission slide 1.

Note: SpectraMax Paradigm Multi-Mode Detection Platform tests designed for a detection cartridge are available if that detection cartridge is installed. Functional Tests are available with the cartridge in the top detection cartridge transport.

TableA-8 Fluorescence Intensity Top Read

Label of Type

Wave-length Alignment Precision Linearity

S/B and S/N

Dynamic Range

Fluorescein 485/535 X X X X X

Rhodamine 535/595 X X X X X

Coumarin 360/465 X

Coumarin B 406/465 X

Texas Red 585/635 X

TableA-9 Fluorescence Intensity Bottom Read

Label of Type


S/B and S/N

Dynamic Range

Fluorescein 485/535 X X

Rhodamine 535/595 X X

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TableA-10 Fluorescence Polarization Read

Label of Type


S/B and S/N

Dynamic Range

Fluorescein 485/535 X

Rhodamine 535/595 X

TableA-11 Fluorescence Time Resolved Read

Label of Type


S/B and S/N

Dynamic Range

Europium Eu/Eu X

Eu-acceptor Eu/665 X

TableA-12 Luminescence Read

Label of Type


S/B and S/N

Dynamic Range

generic no filter X X X X X

TableA-13 Absorbance Read

Label of Type


Accuracy

OD Wave-length

generic fixed X X X

generic 405 X X X X

genomic fixed X X X

generic scan X

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