Molecular Devices FlexStation 3 Manuel utilisateur

www.moleculardevices.com

FlexStation® 3 Benchtop Multi-Mode

Microplate Reader

User Guide

0112-0127 B

June 2013

FlexStation_manual.book Page 1 Tuesday, June 25, 2013 10:22 AM

This document is provided to customers who have purchased 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 non-

transferrable license. 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.

Each product is shipped with documentation stating specifications and other

technical information. Molecular Devices products are warranted to meet the stated

specifications. Molecular Devices makes no other warranties or representations

express or implied, including but not limited to, the fitness of this product 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. The

sole obligation of Molecular Devices and the customer's sole remedy are limited to

repair or replacement of the product in the event that the product fails to perform as

warranted.

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

Product manufactured by Molecular Devices, LLC.

1311 Orleans Drive, Sunnyvale, California, United States of America 94089.

Molecular Devices, LLC is ISO 9001 registered.

The trademarks mentioned herein are the property of Molecular Devices, LLC 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, LLC.

Patents: http://www.moleculardevices.com/productpatents

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0112-0127 B 3

Contents

Chapter 1: Description. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

How To Use This User Guide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

User Guide Organization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

Safety Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

Conventions Used for Precautionary Information. . . . . . . . . . . . . 10

Electrical Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

Service-Trained Users . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

Avoiding Mechanical Problems During Fluid Transfer . . . . . . . . . 11

Safety Messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

System Overview. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

System Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15

Covers and Instrument Panels. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

Drawers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

Fluidics Module. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

Detection Module . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

Computer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28

Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28

Consumables. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

Overview of Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

Choosing an Experiment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

Preparing the Instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

Preparing the Software . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Running the Experiment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Analyzing the Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Theory of Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Instrument Design . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Assay Read Types . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38

SoftMax Pro Software. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

Set Up the Reader and Software Parameters . . . . . . . . . . . . . . . . 42

Acquire Data from the Reader . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43

Perform Complex Data Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . 44

User Interface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45

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FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide

4 0112-0127 B

Chapter 2: Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49

General Precautionary Information . . . . . . . . . . . . . . . . . . . . . . . . . . 49

Unpacking the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50

Unpacking the Fluidics Module and Accessories . . . . . . . . . . . . . . 51

Unpacking the Detection Module . . . . . . . . . . . . . . . . . . . . . . . . . . 51

Removing the Shipping Screws . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52

Installing the Fluidics Module . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53

Installing the Pipettor Head . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56

Setting Up the Computer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61

Connecting the Cables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61

Installing the Drawer Adapters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62

Microplate Adapter Installation. . . . . . . . . . . . . . . . . . . . . . . . . . . . 62

Compound Baseplate Installation . . . . . . . . . . . . . . . . . . . . . . . . . . 63

Installing SoftMax Pro . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63

Chapter 3: Operating Procedures . . . . . . . . . . . . . . . . . . . . . . . . 65

Overview. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66

Starting Up the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67

Using the Control Panel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70

Setting the Temperature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72

Displaying the Temperature. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72

Setting the Temperature with the Control Panel . . . . . . . . . . . . . . 73

Setting the Temperature with SoftMax Pro Software . . . . . . . . . . 74

Setting Up the Software . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75

SoftMax Pro Software Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . 78

Read Types. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79

Read Modes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79

Wavelengths . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80

Sensitivity. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80

Timing. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81

Assay Plate Type . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82

Wells to Read. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83

Automix . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84

AutoCalibrate. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85

Settling Time . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85

AutoRead . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85

SoftMax Pro Software Parameters for Fluid Transfer. . . . . . . . . . . . 86

Compound Source. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87

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Contents

0112-0127 B 5

Compound Transfer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 88

Triturate Selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92

Pipette Tips Layout. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93

Compound and Tip Columns . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 93

Pipette Tip Air Gap . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97

Settings Displayed in Plate Sections . . . . . . . . . . . . . . . . . . . . . . . . . 98

Other Software Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99

Reading a Microplate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99

Loading Tips and Microplates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99

Starting the Reading. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 100

Selecting a Plate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101

Replacing Data in a Plate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101

Viewing Experiment Progress . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102

Data Display . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102

Shutting Down the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103

Optimizing Fluorescence Assays . . . . . . . . . . . . . . . . . . . . . . . . . . . 103

Using Spectral Scanning to Optimize Excitation and Emission

Wavelengths for Fluorescence Assays . . . . . . . . . . . . . . . . . . . . . 105

Optimizing Time-Resolved Fluorescence Assays . . . . . . . . . . . . . . 110

Optimizing Fluorescence Polarization Assays . . . . . . . . . . . . . . . . . 111

Optimizing Luminescence Assays . . . . . . . . . . . . . . . . . . . . . . . . . . 112

Chapter 4: Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113

Obtaining Support. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114

Moving the Instrument. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114

Cleaning the Instrument. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 115

Cleaning Up Spills . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116

Cleaning the Fan Filter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116

Cleaning the Barrels on the Pipettor Head. . . . . . . . . . . . . . . . . . 117

Using the Microplate Adapters . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120

Microplate Adapter Installation . . . . . . . . . . . . . . . . . . . . . . . . . . 120

Removing the Microplate Adapter . . . . . . . . . . . . . . . . . . . . . . . . 120

Using the Compound Baseplate. . . . . . . . . . . . . . . . . . . . . . . . . . . . 121

Replacing Fuses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121

Replacing the Flash Lamp . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123

Long-Term Shutdown . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 128

Chapter 5: Troubleshooting Procedures. . . . . . . . . . . . . . . . . . 129

Problems During Startup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130

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FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide

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Opening a Drawer Manually . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 132

Understanding Potential Mechanical Problems . . . . . . . . . . . . . . . 133

Background . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133

Before Using the Instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133

Avoiding Mechanical Problems . . . . . . . . . . . . . . . . . . . . . . . . . . . 134

In Case of Power Failure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135

Recovering from Mechanical Problems in Flex Mode when Using

Fluidics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135

Assessing a Mechanical Problem . . . . . . . . . . . . . . . . . . . . . . . . . . 136

Opening the Instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137

Evaluating the Tip Rack. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 138

Inspecting Inside the Fluidics Module . . . . . . . . . . . . . . . . . . . . . . 139

Removing the Pipettor Head . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 142

Expelling Undispensed Fluid from Tips . . . . . . . . . . . . . . . . . . . . . 145

Recovery Procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 146

General Error Messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 147

Error Messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 147

Other Error Messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .148

Tilting or Removing the Fluidics Module . . . . . . . . . . . . . . . . . . . . . 149

Tilting the Fluidics Module . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149

Removing the Fluidics Module. . . . . . . . . . . . . . . . . . . . . . . . . . . . 151

Appendix A: Parts and Accessories . . . . . . . . . . . . . . . . . . . . . . 153

Ordering Parts and Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154

Appendix B: Performance Specifications . . . . . . . . . . . . . . . . . 155

Glossary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163

Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167

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0112-0127 B 7

Description

This chapter provides background information on the system, including

descriptions of the principal components and overviews of how the

system functions. It is divided into the following sections:

•Introduction, see page 7

•How To Use This User Guide, see page 9

•Safety Information, see page 9

•System Overview, see page 13

•System Components, see page 15

•Overview of Operation, see page 32

•Theory of Operation, see page 33

•SoftMax Pro Software, see page 41

Introduction

The FlexStation® 3 Benchtop Multi-Mode Microplate Reader combines

the performance of the Molecular Devices SpectraMax® M5e Multi-

Mode Microplate Reader with an integrated 8-channel or 16-channel

pipettor into one compact benchtop reader. This integrated system

provides users with a multi-detection platform capable of increasing the

liquid handling throughput and flexibility for biochemical-based and cell-

based assays. Using an integrated 8-channel or 16-channel pipettor

increases assay flexibility by transferring reagents from 96 or 384 distinct

wells in a source plate to the read plate. This method enables users to

define individual reagents or compound concentrations to be delivered

to each well during the assay. The direct transfer from a source

microplate reduces consumption and allows more assay conditions to be

explored in a single microplate, making the system equally amenable to

agonist and antagonist assay formats. These additions can either occur

concurrently with kinetic analysis of reactions or before an assay to

automate reagent additions. Combined fluid transfer with multi-

detection optics provides a single reader capable of performing a broad

span of applications that pass through drug discovery and research

environments.

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FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide

8 0112-0127 B

Based on the SpectraMax M5e instrument platform, the FlexStation 3

instrument can address detection modalities including absorbance,

fluorescence intensity, fluorescence polarization, time-resolved

fluorescence, and luminescence. Dual monochromators allow users to

target the optimal assay excitation and emission wavelengths, while

eliminating the need to change expensive band pass filters between

experiments. Dual PMTs are integrated into the system to provide

flexibility to detect multiple detection modes, while a separate PMT

provides additional sensitivity for luminescence applications. Reference

diodes automatically adjust to slight fluctuations in excitation intensity to

reduce measurement noise. Absorbance applications are enhanced using

top-quality UV-grade fibers to provide high light transmission in the

lowest wavelengths. These optical characteristics enable the system

performance to be comparable to a top-of-the-line dedicated

spectrophotometer or spectrofluorometer with no trade-off between

instrument performance and the number of read modes.

Figure 1-1: FlexStation 3 Instrument

FlexStation_manual.book Page 8 Tuesday, June 25, 2013 10:22 AM

Description

0112-0127 B 9

How To Use This User Guide

This user guide was written to ensure safe and proper use of the system.

Before use, read this user guide carefully to realize the full capabilities of

the system. Also, if something is unclear during daily use or if a problem

occurs, please refer to this user guide.

User Guide Organization

This user guide is organized as follows:

•Description on page 7 provides background information on the

system, including component descriptions, functional overviews,

and safety information.

•Installation on page 49 provides instructions for setting up the

reader.

•Operating Procedures on page 65 provides instructions for

starting up the reader, setting parameters for the various read

modes, and reading a microplate.

•Maintenance on page 113 provides instructions for cleaning the

fan filter, changing the fuses, and moving the system to a new

location.

•Troubleshooting Procedures on page 129 provides instructions

for diagnosing and solving common problems, as well as a list of

error conditions.

•Parts and Accessories on page 153 provides a list of spare parts.

•Performance Specifications on page 155 provides the technical

specifications for the instrument.

•Glossary on page 163 provides a list of terms and definitions.

Safety Information

When operated properly in a safe environment and according to the

instructions in this user guide, there are no known hazards associated

with the FlexStation 3 instrument. However, proper use requires an

understanding of situations that are potentially dangerous and can result

in serious injury. All users must be familiar with the guidelines in this

section before working with the system.

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FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide

10 0112-0127 B

Conventions Used for Precautionary Information

This user guide uses the following conventions to provide technical and

safety information of special interest.

Electrical Safety

Note: A note gives background information that is provided to clarify a

particular step or procedure. A not can also provided an instruction to

ensure correct results and optimal performance.

CAUTION! A caution is an instruction that, if not followed, can result in

damage to the system or in loss of data.

WARNING! A warning is an instruction that, if not followed, can result

in potential injury to a person working with the system.

WARNING! BIOHAZARD. A biohazard warning indicates a condition

involving potentially infectious biological agents requiring that proper

handling precautions be taken.

WARNING! Follow all instructions in this user guide and on system

labels. If you use the system in a manner not specified by Molecular

Devices, then any protections provided by the system might be

impaired.

FlexStation_manual.book Page 10 Tuesday, June 25, 2013 10:22 AM

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