Molecular Devices FlexStation 3 User Manual
FlexStation® 3 Benchtop Multi-Mode
Microplate Reader
User Guide
0112-0127 B
June 2013
www.moleculardevices.com
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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
0112-0127 B 3
FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide
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
4 0112-0127 B
Contents
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
0112-0127 B 5
FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide
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
6 0112-0127 B
Description
Introduction
1
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
The FlexStation® 3 Benchtop Multi-Mode Microplate Reader combines
the performance of the Molecular Devices SpectraMax® M5e MultiMode 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 cellbased 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 multidetection optics provides a single reader capable of performing a broad
span of applications that pass through drug discovery and research
environments.
0112-0127 B 7
FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide
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
8 0112-0127 B
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.
Description
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.
0112-0127 B 9
FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide
Conventions Used for Precautionary Information
This user guide uses the following conventions to provide technical and
safety information of special interest.
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.
Electrical Safety
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.
10 0112-0127 B
Description
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:
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.
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.
0112-0127 B 11
FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide
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
12 0112-0127 B
System Overview
The FlexStation 3 Benchtop Multi-Mode Microplate Reader is a
monochromator-based microplate reader that has 6-well, 12-well, 24well, 48-well, 96-well, and 384-well microplate reading capability for
absorbance, fluorescence intensity, fluorescence polarization, timeresolved fluorescence, and luminescence. When using the integrated
pipettor, the instrument offers 96-well and 384-well microplate-tomicroplate 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.
Description
0112-0127 B 13
FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide
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.
14 0112-0127 B
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
Description
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
0112-0127 B 15
FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide
Figure 1-3: Instrument, Front View
Figure 1-4: Instrument, Rear View
16 0112-0127 B
Description
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.
Note: The top cover can be lifted back, as shown in the figure below,
for certain limited troubleshooting procedures. See
Instrument on page 137.
Opening the
Figure 1-5: Lifting Off the Top Cover
Note: To achieve optimal performance during readings, you must
operate the system with the top cover in place.
0112-0127 B 17
FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide
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
Starting Up the System on page 67 and Setting the Temperature on
See
page 72.
18 0112-0127 B
Description
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
0112-0127 B 19
Connecting the Cables on page 61.
FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide
Lamp Cover
The plastic lamp cover provides access to the flash lamp on the right side
of the instrument (as viewed from the rear).
CAUTION! Flash lamp access and maintenance are restricted to servicetrained users.
Figure 1-8: Rear View
Replacing the Flash Lamp on page 123.
See
20 0112-0127 B
Description
Drawers
The instrument has three drawers that open on the right side. The two
drawers in the fluidics module open and close to move the pipette tip
rack and compound plates (or reservoirs) into and out of the instrument.
The reading chamber drawer in the detection module transports the
assay microplate into the reading chamber.
Figure 1-9: Instrument with Drawers Open and Carriages Accessible
Small plastic pushers, in the front left corner of each drawer, hold the
plates, racks, or reservoirs securely in place when the drawers are closed.
Figure 1-10: Drawer Detail
0112-0127 B 21
FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide
CAUTION! Do not obstruct the movement of any of the drawers. If you
must retrieve a plate after an error condition or power outage, and if the
drawer will not open, it is possible to open the drawer manually. See
Opening a Drawer Manually on page 132.
You can open and close the drawers using either the SoftMax Pro
Software or by pressing the drawer keys on the instrument control panel.
Using the SoftMax Pro Software, open the Control menu and click Tip
Drawer for the tip rack drawer, Compound Drawer for the compound
plate drawer, or Open Drawer for the reading chamber drawer.
Figure 1-11: SoftMax Pro Software Control Menu
You can also open or close the reading chamber drawer with the Drawer
button on the Status Bar.
Figure 1-12: SoftMax Pro Software Drawer Button
22 0112-0127 B
Description
Tip Rack Drawer
The top drawer holds the pipette tip rack.
Only tips specified by Molecular Devices for use with the FlexStation 3
instrument can be safely used. See
CAUTION! Do not use parts and accessories that are not authorized by,
specified by or provided by Molecular Devices. Using unauthorized parts
can damage the instrument.
Parts and Accessories on page 153.
Compound Plate Drawer
The compound plate drawer holds a 96-well or 384-well microplate. The
instrument can simultaneously transfer a column of fluids from the
compound plate:
• Eight fluids from a 96-well compound plate to a 96-well assay
plate
• Sixteen fluids from a 384-well compound plate to a 384-well
assay plate
Note: Be sure to install the compound baseplate before placing a
compound plate in the drawer.
Reading Chamber Drawer
The reading chamber drawer opens to accept a 96-well and 384-well
microplate for analysis in the reading chamber. It is the lowest of the
three drawers.
The reading chamber drawer operation varies, depending on the
incubator status. When the incubator is off, the reading chamber drawer
is open at power up and after a read. When the incubator is on, the
drawer closes automatically to maintain the temperature of the reading
chamber.
Note: Be sure to install the black microplate adapter before placing an
assay plate in the drawer for standard 96-well and 384-well microplates.
See Installing the Drawer Adapters on page 62.
0112-0127 B 23
FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide
Fluidics Module
The fluidics module houses the pipettor head, several motors, and all the
fluidics components. There are two horizontally-moving carriers, one for
the pipette tip rack and the other for the compound plate. The pipettor
head moves vertically between the drawers.
The fluidics module can be opened, by service-trained users, from the
inside front panel, if necessary for maintenance, or from the inside top
panel to install or remove the pipettor head. The entire fluidics module
can be removed for maintenance or to transport the system to another
location.
Figure 1-13: Fluidics Module
Installing the Fluidics Module on page 53 or See Troubleshooting
See
Procedures on page 129.
24 0112-0127 B
Description
Pipettor Head
The instrument is configured with an 8-channel pipettor head for use
with 96-well microplates or a 16-channel pipettor head for use with 384well microplates.
Figure 1-14: Pipettor Head
Installing the Pipettor Head on page 56.
See
The barrels on the pipettor head require periodic cleaning to remove
silicone lubricant, dust, and other miscellaneous contamination. See
Cleaning the Barrels on the Pipettor Head on page 117.
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FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide
Detection Module
The detection module is the lower portion of the instrument. This
module houses the reading chamber, the optics bench, several cables
and optic fibers, the power supply, the flash lamp, and other hardware.
The fluidics module attaches to the detection module and can be tilted
off to the side, to provide access to the optical system for
troubleshooting or maintenance. The detection module is contained in a
molded plastic housing, to which the top cover is attached at the back of
the instrument.
Figure 1-15: Detection Module Detail
Reading Chamber
The reading chamber includes the assay plate carriage that holds the
assay microplate in the reading chamber during read cycles. The reading
chamber can be maintained at an elevated temperature. It contains both
top and bottom read heads that can be selected in the software.
The instrument uses a plate sensor to assure that an assay plate is
present in the reading chamber before a reading begins.
26 0112-0127 B
Description
Optical System
The optical system includes a xenon flash lamp, monochromators,
excitation bandpass filters, emission cut-off filters, PMTs, and
photodiodes.
There are a number of cables and fibers that exit the optics bench and
enter the reading chamber. They are the excitation fibers (thin and black
or red, has a collar and pins), emission fibers (black and fatter, with
attached electrical cord), electrical connector to the read head (green
with brass fitting).
Figure 1-16: Optical System
CAUTION! Optical fibers are very fragile, especially the excitation fiber.
Handle cables with extreme care. Do not flex, twist, bend, or stretch the
optical cables.
0112-0127 B 27
FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide
Computer
The FlexStation 3 instrument works as a system with the SoftMax Pro
Software. The SoftMax Pro Software must be installed on a dedicated
computer to communicate with and control instrument functions.
The instrument is equipped with an 8-pin DIN RS-232 serial port for
connecting to a computer.
SoftMax Pro Software, version 5.1 or later, is required to control the
FlexStation 3 instrument. TheFlexStation 3 instrument is not currently
supported in SoftMax Pro Software, version 6.x.
The minimum computer configuration includes a Pentium processor with
2.8 GHz, 1 GB hard drive.
See
Installing SoftMax Pro on page 63 and Setting Up the Software on
page 75.
Accessories
The following accessories are included with the system:
• Black microplate adapter (for use in reading chamber drawer)
• Compound baseplate (for use in the compound plate drawer)
• Computer cable
• Power cord, USA/Canada
• Power cord, ECI
• Fuses (2 each)
• User Guide
• Pipettor head, 8-channel and/or 16-channel
• Pipette
• Yellow plate for the respective pipettor head
• Hex key
All necessary accessories are shipped with the system.
Accessories on page 153.
Fuses are rated slow-blow (United States/Canada/Metric: 6.3 amp time
delay). See
Replacing Fuses on page 121.
Parts and
28 0112-0127 B
Description
Cables
Molecular Devices recommends that you use high-quality, doubleshielded cables to connect the instrument to the computer. Choose
cables that meet the following requirements:
Serial Interface Cable: The serial interface cable used to connect the
instrument to the computer is a custom cable designed and built by
Molecular Devices. Use the cable supplied by Molecular Devices, or
contact Molecular Devices for specific pin-out requirements: Male DB8
to Female DB9 (custom cable made by Molecular Devices,
P/N 9000-0149).
USB Adapter Cable: Many newer computers do not have a serial port.
You can connect a serial cable between these computers and the
instrument using a USB-to-serial adapter. Molecular Devices has tested
many third-party USB-to-serial adapter cables and has found Keyspan
USA-19HS (Molecular Devices, P/N 9000-0938) to be the most reliable. It
is the only one we recommend.
Note: For specific pin-out requirements, contact Molecular Devices
Technical Support.
Microplate Adapters
The black microplate adapter fits in the assay plate carriage, in the
reading chamber drawer, to elevate standard microplates for both top
reads and bottom reads. Remove the adapter when using high-profile,
6-well, 12-well, 24-well, or 48-well microplates.
Compound Baseplate
Molecular Devices provides a metal baseplate that must be placed in the
compound plate drawer under the compounds plate to reduce stray
light.
0112-0127 B 29
FlexStation 3 Benchtop Multi-Mode Microplate Reader User Guide
Consumables
The system consumables include:
• Microplates
• Pipette tips
One box (10 racks) of pipette tips is shipped with the system pipettor
head. See
CAUTION! Do not use parts and accessories that are not authorized by,
specified by, or provided by Molecular Devices. Using unauthorized parts
can damage the instrument.
Microplates
The FlexStation 3 instrument can accommodate standard 6-well to
384-well microplates and strip wells. When reading absorbance at
wavelengths below 340 nm, special UV-transparent, disposable or quartz
microplates allowing transmission of the deep UV spectra must be used.
Not all manufacturers’ microplates are the same with regard to design,
materials, or configuration. Temperature uniformity within the
microplate can vary depending on the type of microplate used.
Microplates supported for use in this reader are:
Read plate formats not already loaded as defaults in the SoftMax Pro
Software can be added by manually entering the dimensions using the
Plate Editor in the software.
The instrument can accommodate standard 6-well, 12-well, 24-well,
48-well, 96-well, and 384-well microplates. In Flex or other assay types
which you intend on using fluidic integration you can only use 96-well or
384-well formatted assay plates.
Parts and Accessories on page 153.
• 6-well, 12-well, 24-well, 48-well, 96-well, and 384-well standard
formats
• 96-well half area
• 96-well and 384-well low volume
30 0112-0127 B
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