Molecular Devices FlexStation 3 User manual

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.
© 2013 Molecular Devices, LLC.
All rights reserved.
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
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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
1
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
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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.
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Description
0112-0127 B 11
Service-Trained Users
There are two types of users described in this user guide. Most
procedures required for operating and troubleshooting can be
performed by any user who has read the instructions in this user guide
and is familiar with the system. However, all installation procedures, and
some more complex service and troubleshooting procedures, require the
expertise of a service-trained user. Whenever the following warning
message appears, a service-trained user must perform the procedure to
ensure user safety and to prevent instrument damage.
Example:
Avoiding Mechanical Problems During Fluid Transfer
Because of the complex mechanical nature of the FlexStation 3
instrument, including both fluidics and optical reading, smooth and
reliable operation of the system depends on both good design and
operator knowledge.
To prevent problems of a mechanical nature, be sure to read all sections
of this user guide before attempting a reading with fluidics. See
Understanding Potential Mechanical Problems on page 133.
Safety Messages
Observe the following warnings and precautions:
High internal voltages. Always turn off the power switch and unplug the
system power cord before removing labeled covers or panels.
Xenon-arc flash lamp. Do not look directly at the flash lamp while it is
illuminated. The lamp emits ultraviolet radiation at levels that can injure
the eye if viewed directly.
Electrical grounding. Never use a two-prong plug or extension cord to
connect primary power to the system. Use of a two-prong adapter
disconnects the utility ground, creating a severe shock hazard. Always
connect the system power cord directly to a three-prong receptacle with
a functional ground.
WARNING! The following procedures must be completed by a
service-trained user. Do not attempt the following procedures if you
have not been trained properly by appropriate Molecular Devices
personnel.
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FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide
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Spilled liquids. Avoid spilling liquids on the system. Fluid spilled into
internal components creates a potential shock hazard. Wipe up all spills
immediately. Do not operate the system if internal components have
been exposed to spilled fluid. Unplug instrument if there is a fluid spill in
the instrument and contact Technical Support.
Replacement fuses. Use replacement fuses with the required current
rating and specification. Improper fuses or short-circuiting the fuse
holders can cause fire or damage the instrument.
Power rating.Ensure the system is connected to a power receptacle that
provides voltage and current within the specified rating for the system.
Use of an incompatible power receptacle can create electrical shock and
fire hazards.
Remove watches and jewelry before removing any panels from the
instrument.
Warning labels. There are several labels affixed to the instrument covers
and inside panels. The purpose of these labels is to alert the user to use
caution when servicing a component or the instrument. The user should
be aware that ignoring the instructions on any instrument label can
result in a hazardous condition that can cause injury.
Identification labels:The following label, among others, appears on the
instrument.
\
Figure 1-2: FlexStation 3 Instrument Label
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Description
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System Overview
The FlexStation 3 Benchtop Multi-Mode Microplate Reader is a
monochromator-based microplate reader that has 6-well, 12-well, 24-
well, 48-well, 96-well, and 384-well microplate reading capability for
absorbance, fluorescence intensity, fluorescence polarization, time-
resolved fluorescence, and luminescence. When using the integrated
pipettor, the instrument offers 96-well and 384-well microplate-to-
microplate fluid transfer, 8 or 16 wells at a time.
The top portion of the instrument, the fluidics module, uses an 8-channel
or 16-channel pipettor, to expand assay flexibility by transferring
reagents from a 96-well or 384-well source plate to the assay plate.
When transferring reagents from distinct wells of a microplate, users can
define individual reagents or concentrations to be delivered in each well
during their assay. This direct transfer allows more assay conditions to be
explored in a single microplate, reducing reagent consumption as well as
making the system more amenable to both agonist and antagonist assay
formats. In addition, kinetic cell-based assay throughput (for example,
calcium mobilization) is increased when 8 or 16 wells of a microplate are
analyzed in conjunction rather than individually.
Integrated pipetting provides flexibility, and also offers users parameters
to optimize assay robustness. Trituration, mixing via aspiration and
dispense of the pipettor, encourages mixing to either resuspend source
plate compounds or spontaneously mix reagents to promote a rapid
response with minimal assay variability. Dispense parameters can also be
optimized for each experiment to accommodate cells with different
adherence characteristics to prevent cell dislodging. Furthermore, the
integrated fluidics platform uses disposable pipette tips to minimize
reagent cross contamination between wells or experiments.
The fluidics module interfaces with the lower reading chamber which
encloses a high-powered Xenon flash lamp as the light source. Sensitivity
or read-speed can be optimized by varying the number of lamp flashes
per read.
The two holographic diffraction grating monochromators allow selection
of any wavelength between 200 nm and 1000 nm in absorbance; 250 nm
and 850 nm in fluorescence intensity, time-resolved fluorescence (TRF),
or luminescence; and 400 nm and 750 nm for readings in fluorescence
polarization. Excitation and emission wavelengths are optimized using
the mirrored optics to focus light into the sample volume, and cutoff
filters to reduce stray light and minimize background interference.
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The system has five assay types available which include Flex, Endpoint,
Kinetic, Spectrum, and multi-point Well Scan. Flex mode assay enables
users to transfer fluid from the source read plate while immediately
reading a fast kinetic assay that uses absorbance, fluorescence, or
luminescence detection technologies. Alternatively, the pipettor head
can be used to transfer fluid before an endpoint or kinetic assay to
automate fluid transfer and minimize user interaction.
In addition to fluid transfer, well contents can be mixed automatically by
shaking before each read cycle, making it possible to perform kinetic
analysis of solid-phase, enzyme-mediated reactions.
Top detection in microplates is available for all fluorescence and
luminescence readings. Bottom detection is available for all assay types
except fluorescence polarization. When reading absorbance at
wavelengths below 340 nm, special UV-transparent, disposable or quartz
microplates that allow transmission of the far ultraviolet spectra must be
used.
A plate drawer adapter is provided with the reader. The adapter is
required for optimum performance when reading from the top or
bottom of standard 96-well and 384-well microplates in all read types,
including absorbance.
Variations in measured fluorescence values are virtually eliminated by
internal compensation for detector sensitivity, photomultiplier tube
voltage, and excitation source intensity.
Using the FlexStation 3 instrument with the PathCheck® Pathlength
Measurement Technology allows normalization of variable well volumes
to absorbance readings. In addition PathCheck technology allows for
pipettor validation, including the online 8-channel and 16-channel
pipettors, and to compare experiments from different days.
Temperature in the microplate chamber is isothermal, both at ambient
and when the incubator is turned on. When the incubator is on, the
reading chamber temperature can be controlled from 2°C above ambient
room temperature to 45°C. Please note that the temperature of the
fluidics module is not regulated and that it is recommend that any
microplates or tips should be at the desired temperature before placing
them inside the instrument.
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Description
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The FlexStation 3 instrument is controlled by an external computer
running the SoftMax® Pro Microplate Data Acquisition and Analysis
Software, which provides integrated instrument control, data display,
and statistical data analysis. The on-board microprocessor calculates and
reports the absorbance, percent transmittance, fluorescence, or
luminescence for each well of a microplate. Data from multiple
wavelengths can be acquired and ratio analysis can be performed during
a single reading, if desired. In addition, different calculations can be
made based on this data using the SoftMax Pro software, including the
subtraction of blanks, quantitation from standard curves, calculation of
IC50 values, and more.
The extreme flexibility and high sensitivity of the FlexStation 3 reader
makes it appropriate for applications within the fields of biochemistry,
cell biology, immunology, toxicology, molecular biology, and
microbiology. Online fluidic integration expands the capabilities of the
instrument to include fast fluorescence (calcium mobilization),
luminescence, and absorbance assays in addition to typical applications
which include ELISA, nucleic acid and protein quantitation,
homogeneous and heterogeneous enzyme-activity assays, and microbial
growth, endotoxin testing, and reporter-gene assays.
System Components
This section describes the major system components listed below.
•Covers and Instrument Panels, see page 17
•Drawers, see page 21
•Fluidics Module, see page 24
•Detection Module, see page 26
•Computer, see page 28
•Accessories, see page 28
•Consumables, see page 30
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Figure 1-3: Instrument, Front View
Figure 1-4: Instrument, Rear View
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Description
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Covers and Instrument Panels
Top Cover
The instrument is protected by a molded plastic housing. The large top
cover protects the fluidics module and the exposed portions of the
detection module.
Figure 1-5: Lifting Off the Top Cover
Note: The top cover can be lifted back, as shown in the figure below,
for certain limited troubleshooting procedures. See Opening the
Instrument on page 137.
Note: To achieve optimal performance during readings, you must
operate the system with the top cover in place.
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Control Panel
The control panel consists of an LCD and six pressure-sensitive
membrane keys which can be used to initiate and regulate the
temperature and to open and close the drawers.
A 2×3-character liquid crystal display (LCD) shows the current instrument
temperature at all times, and the set point temperature when the
incubator is on.
Figure 1-6: Control Panel
See Starting Up the System on page 67 and Setting the Temperature on
page 72.
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Description
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Input/Output Panels
There are two input/output panels on the rear of the instrument.
•The upper input/output panel, on the back cover, consists of a
power switch, fuse box cover, and power cord receptacle.
•The lower panel consists of an RS-232 serial port and parallel
port (not currently active). There are also a number of
identification labels.
Figure 1-7: Input/Output Panels.
For information about attaching the computer cable and power cords to
the instrument, see Connecting the Cables on page 61.
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Lamp Cover
The plastic lamp cover provides access to the flash lamp on the right side
of the instrument (as viewed from the rear).
Figure 1-8: Rear View
See Replacing the Flash Lamp on page 123.
CAUTION! Flash lamp access and maintenance are restricted to service-
trained users.
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