Molecular Devices ImageXpress Pico User manual

5063666 F
May 2019
ImageXpress™Pico
Automated Cell Imaging System
with CellReporterXpress™ Software Version 2.1
Installation Guide

ImageXpress Pico Personal Imaging System Installation Guide
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©2019Molecular Devices, LLC.
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Contents
Safety Information 5
Warnings, Cautions, Notes, and Tips 5
Symbols on Instrument Labels 6
Protective Housing 7
Electrical Safety 8
Moving Parts Safety 10
Lifting Hazard 10
Chemical and Biological Safety 11
Chapter 1: ImageXpressPicoAutomated Cell Imaging System 13
ImageXpressPico System Features 14
CellReporterXpress Software Features 16
Theory of Operation 17
Obtaining Support 21
Product Documentation 22
About This Guide 22
Chapter 2: Setting Up the ImageXpressPico System 23
Package Contents 23
Environmental Requirements 25
Space and Table Requirements 26
Power Requirements 27
Unpacking the Instrument 28
Placing the Instrument 31
Removing the Transport Lock 31
Connecting Cables 32
Powering On the Instrument 34
Chapter 3: Installing the CellReporterXpress Software 35
Chapter 4: Finishing Setup of the ImageXpressPico System 37
Logging In to the Software 37
Installing an Objective 38
Installing a Filter Cube 40

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Inserting the Plate Holder or Slide Holder 42
Chapter 5: Environmental Control System 45
Environmental Control System Warnings and Cautions 46
Environmental Control System Hardware 47
Using the Environmental Control System 50
Appendix A: Instrument Specifications 59
Instrument Dimensions without Environmental Control 61
Instrument Dimensions with Environmental Control 64
Appendix B: Optional Parts and Accessories 67
Objectives 67
Filter Cubes 68
Accessories 68
Appendix C: Environmental Control System Gas Regulator Details 69
Using a Gas Cylinder 70
Using a Lab Gas Line 72
Using an Oil-Free Air Compressor 74
Unsupported Gas Supplies 74
Appendix D: Packing the Instrument for Transport 75
Appendix E: Electromagnetic Compatibility 81

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Safety Information
This section describes the safe use of the instrument. Safety includes an understanding of
the information in this guide, the safety labels on the instrument, and the precautions that
you must follow before and during operation of the instrument.
It is important to confirm that everyone involved with the operation of the instrument has:
Received instruction in general safety practices for laboratories.
Received instruction in specific safety practices for the instrument.
Read and understood all Safety Data Sheets (SDS) for all materials being used.
Read and understood all instrument documentation, including all warnings and
cautions.
Always remember that the key to safety is operating the instrument with care.
WARNING! If the instrument is used in a manner not specified by Molecular
Devices, the protection provided by the equipment might be impaired.
Warnings, Cautions, Notes, and Tips
All warning symbols in the user guide are framed within a yellow triangle. An exclamation
mark is used for most warnings. Other symbols can warn of other types of hazards such as
biohazard, electrical, or laser safety warnings as are described in the text of the warning.
When warnings and cautions are displayed in this guide, be careful to follow the specific
safety information related to them.
The following user-attention statements can be displayed in the text of Molecular Devices
user documentation. Each statement implies a particular amount of observation or
recommended procedure as described:
WARNING! A warning indicates a situation or operation that could cause
personal injury if precautions are not followed. The warning symbol can vary
depending on the warning. The definition of the symbol is included in the text of
the warning.
CAUTION! A caution indicates a situation or operation that could cause damage to
the instrument or loss of data if correct procedures are not followed.
Note: A note calls attention to significant information.
Tip: A tip provides useful information or a shortcut, but is not essential to the
completion of a procedure.

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Symbols on Instrument Labels
Each safety label found on the instrument contains an alert symbol that indicates the type of
potential safety hazard related to the label. The following table lists the alert symbols that
can be found on Molecular Devices instruments.
Symbol Indication
Indicates that the product documentation must be consulted.
Indicates a potential lifting hazard. For information about the weight of the instrument,
Indicates a potential heat hazard.
Indicates a rotating parts hazard.
Indicates the location of a fuse.
Indicates the instrument serial number.
Indicates the instrument manufacture date.
Indicates that you should consult the instructions for use.
Indicates CSA certification.
Indicates European technology conformity.

Safety Information
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Symbol Indication
Indicates compliance with Australian radio communication requirements.
Indicates compliance with the Waste Electrical and Electronic Equipment (WEEE)
Directive of the European Union.You must not discard this electrical or electronic
product or its components in domestic household waste or in the municipal waste
collection system.
For products under the requirement of the WEEE directive, contact your dealer or
local Molecular Devices office for the procedures to facilitate the proper collection,
treatment, recovery, recycling, and safe disposal of the device.
Indicates the environmental friendly use period for China RoHS. The symbol may
indicate the number of years in the use period.
Indicates that there is an authorized representative in the European community.
Indicates the instrument manufacturer.
Indicates the manufacturer catalog number.
Protective Housing
The instrument features a protective outer housing, which is designed to protect you from
exposure to LED light, hot surfaces, and moving parts.
WARNING! Do not manually open the plate door, open the protective housing,
or attempt to gain access to the interior of the instrument. These actions can
damage the instrument components and result in hazardous exposure to LED light,
hot surfaces, moving parts, or high voltage.

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Electrical Safety
To prevent electrically related injuries and property damage, inspect all electrical equipment
before use and immediately report all electrical deficiencies. Contact Molecular Devices
Technical Support to service equipment that requires the removal of covers or panels.
WARNING! HIGH VOLTAGE. Within the instrument is the potential of an
electrical shock hazard existing from a high-voltage source. Read and understand
all safety instructions before you install, maintain, or service the instrument.
Do not remove the instrument covers. To prevent electrical shock, use the supplied power
cords only and connect to a properly grounded wall outlet.
To ensure sufficient ventilation and provide access for disconnecting power from the
instrument, maintain a clearance of 20 cm to 30 cm (7.9 in. to 11.8 in.) at the rear of the
instrument.
Power off the instrument when not in use.
If the instrument does not power on, you may need to replace the instrument fuses. See
Replacing Fuses on page 9 for details.

Safety Information
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Replacing Fuses
If the instrument does not seem to get power after you press the Power button, confirm
that the power cord is securely connected to a functioning power outlet and to the power
port on the rear of the instrument.
If the power failed while the instrument was running, verify that the power cord is not loose
or disconnected and that power to the power outlet is functioning properly.
If these checks fail to remedy the loss of power, replace the fuses. See Instrument
Specifications on page 59 for fuse details.
CAUTION! Do not touch or loosen screws or parts other than those specifically
designated in the instructions. Doing so could cause misalignment and possibly void
the warranty.
The fuses are located in the fuse carrier, which is below the power port at the rear of the
instrument.
WARNING! HIGH VOLTAGE Always power off the instrument and disconnect the
power cord from the main power source before you perform a maintenance
procedure that requires removal of a panel or cover or disassembly of an interior
instrument component.
To replace fuses:
1. Press and hold the Power button to power off the instrument.
2. Disconnect the power cord from the power port.
3. Gently press on the carrier-release tab (below the power port) with a small flat-head
screwdriver,and pull the fuse carrier to remove it from the instrument.
4. Gently pull the old fuses from the carrier by hand.
5. Gently place the new fuses into the carrier.
6. Press the fuse carrier into the instrument until the carrier snaps into place.
7. Connect the power cord into the power port.
8. Press the Power button to power on the instrument.
Note: If the instrument still does not power on after you change the fuses, contact
Molecular Devices Technical Support. See Obtaining Support on page 21 for details.

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Moving Parts Safety
The instrument contains moving parts that can cause injury. Under normal conditions, the
instrument is designed to protect you from these moving parts.
To prevent injury:
Never try to exchange labware, reagents, or tools while the instrument is operating.
Never try to physically restrict the moving components of the instrument.
WARNING! Do not attempt to access the interior of the instrument unless
specifically instructed to do so. The moving parts inside the instrument can cause
injury. Do not operate the instrument with any covers or panels removed.
Note: Observe all warnings and cautions listed for all external devices attached to or in
use during the operation of the instrument. See the applicable user guide for the
operating and safety procedures of that device.
Lifting Hazard
WARNING! LIFTING HAZARD. The ImageXpressPico system weighs
approximately 38 kg (84lb). Use great care when lifting or moving the instrument.
To prevent injury, use a minimum of two people to lift the instrument.
CAUTION!
Always lift the instrument to move it. Do not slide or push the instrument. Sliding
or pushing can damage the feet on the bottom of the instrument.
Moving the instrument can damage sensitive parts and disrupt optical
alignments. When transporting the instrument, use the original packaging and
shipping box to properly secure the instrument. Your warranty does not cover
problems caused during or as a result of shipment or relocation.

Safety Information
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Chemical and Biological Safety
Normal operation of the instrument can involve the use of materials that are toxic,
flammable, or otherwise biologically harmful. When using such materials, observe the
following precautions:
Handle infectious samples based on good laboratory procedures and methods to
prevent the spread of disease.
Observe all cautionary information printed on the original containers of solutions before
their use.
Dispose of all waste solutions based on the waste disposal procedures of your facility.
Operate the instrument in accordance with the instructions outlined in this guide, and
take all the required precautions when using pathological, toxic, or radioactive materials.
Splashing of liquids can occur. When working with potentially hazardous liquids, take
applicable safety precautions, such as wearing safety glasses and protective clothing.
Observe the applicable cautionary procedures as defined by your safety officer when
using hazardous materials.
Use a compressed gas supply in a well-ventilated area. The instrument is not air-tight.
Gas can escape into the atmosphere surrounding the instrument. When you use
potentially toxic gas, observe the cautionary procedures your safety officer defines to
maintain a safe work environment, such as the use of an automatic warning system.
Observe the applicable cautionary procedures as defined by your safety officer when
using flammable solvents in or near a powered-up instrument.
Observe the applicable cautionary procedures as defined by your safety officer when
using toxic, pathological, or radioactive materials.
WARNING! Never use the instrument in an environment where potentially
damaging liquids or gases are present.

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Chapter 1: ImageXpressPicoAutomated Cell Imaging
System
The Molecular Devices®ImageXpress™ PicoAutomated Cell Imaging System is an affordable
all-in-one platform for automatically acquiring and analyzing images from fluorescently
labeled biological samples in plates and slides. It enables you to increase the throughput of
your image acquisition and analysis, allowing you to gain insights in minutes. With the
addition of a modular option, the system provides environmental control for live cell
imaging.
The core hardware component of the imaging system is a custom-designed, fully automated,
epi-illumination fluorescence microscope. The rapid autofocus and precision sample
movement features of the microscope allow large numbers of high-resolution images to be
acquired in the shortest possible time. All key optical and mechanical elements are
motorized, which allows complete control of the instrument configuration.
When used in combination with the CellReporterXpress™ Image Acquisition and Analysis
Software, which features powerful image analysis capabilities and more than 25 available
predefined experimental protocols, the ImageXpressPico system becomes an extremely
flexible device, ideally suited for user-defined, automated assays.
Key components of the instrument include the following:
Built-in, internal light source comprised of six high-powered LEDs enables very high
sensitivity fluorescent imaging. Additional LED-based light sources allow transmitted light
and overview imaging.
High-sensitivity, 5-megapixel CMOS camera.
Hardware-based autofocus system with precision motorized Z-stage focus.
High-transmission fluorescence imaging optics with world-class chromatic aberration
correction, resolution, and image flatness.
Secondary camera provides overview imaging for slides.
Precision motorized sample (X-Y) stage.
High-quality Leica objectives in a six-position turret.
Filter cubes in a six-position turret.
Motorized selection of stage position, filter cubes, and objectives.
Temperature control up to 40°C (104° F) for live cell imaging.
Optional environmental control system, which enables you to regulate humidity, CO2,
and O2inside the environmental control cassette for multi-day, live-cell, time-lapse
imaging.
Operation and configuration control by the integrated CellReporterXpress software.
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ImageXpressPico System Features
Illumination System
Light Source
The ImageXpressPico system light source is comprised of six high-powered LEDs with a rated
lifetime of more than 20,000 hours. It has an excitation spectrum ranging from ultraviolet to
red. There are no user-replaceable parts in the light source.
Filter Cube Turret
The six-position filter cube turret uses Leica filter cubes, which contain filters that are adapted
and matched to the ImageXpressPico system. Additional filter cubes are available exclusively
from Molecular Devices.
See Filter Cubes on page 68 for details.
Objective (Z) Stage
The Z stage position features a resolution of better than 0.25 μm.
The ImageXpressPico system uses Fluotar objectives manufactured by Leica Microsystems.
The selected objective lens focuses excitation light onto the sample and collects fluorescent
light emitted by the sample.
One or more objectives is included with your initial purchase of the ImageXpressPico
system. After that, you can order additional compatible objectives exclusively from
Molecular Devices. The following objectives are compatible with the ImageXpressPico
system:
Magnification Numerical Aperture Color Band
4x 0.13 Red
10x 0.32 Yellow
20x 0.40 Light Green
40x 0.60 Light Blue
63x 0.70 Dark Blue
CAUTION! To prevent damaging both the instrument and your samples, do not use
any other objectives with the ImageXpressPico system.
See Objectives on page 67 for details.
Motorized Objective Turret
The ImageXpressPico system includes a six-position objective turret.

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Sample (X-Y) Stage
Plate Holder
The plate holder is designed for scanning multi-well microplates in standard ANSI (SBS)
formats with plastic or glass bottoms. It can accommodate other plate formats that have
standard footprint dimensions. Optimal image quality depends on plate flatness, well
bottom thickness, and optical clarity.
A spring-loaded, mechanical clamp holds the sample plate securely in the plate holder. The
clamp automatically opens when the sample stage moves to the load/eject position; it
automatically closes when the sample stage moves the plate into position for imaging.
Slide Holder
The slide holder holds four standard microscope slides of 25 mm x 75 mm (1 in. x 3 in.).
LED Autofocus
The ImageXpressPico system uses a high-powered LEDto autofocus the sample. For certain
assays (particularly those with thin plates and low magnifications), software autofocus is also
available.

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CellReporterXpress Software Features
The CellReporterXpressImage Acquisition and Analysis Software by Molecular Devices is the
user interface for the ImageXpressPico system. You will use the CellReporterXpress software
to work with the ImageXpressPico system and control all its functions.
The CellReporterXpress software integrates image acquisition and analysis into a unified
workflow. Along with the ImageXpressPico system, the CellReporterXpress software
streamlines automated imaging to offer a simplified solution for scaling up microscopy. Its
features include:
A web-based interface that runs on many browsers, including those found on iPads and
Android tablets.
Over 25 available predefined experimental protocols.
High-powered analysis tools equivalent to those found in desktop applications.
Easy-to-manage data with no requirement to configure a database.
A simplified user interface that is easy to learn and easy to use.
As part of setting up the ImageXpressPico system, you will install the CellReporterXpress
software.
Note: See the CellReporterXpress Help or the CellReporterXpress User Guide for
details on using the CellReporterXpress software.

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Theory of Operation
The ImageXpressPico system uses the following components and functions:
Fluorescence Imaging, see below
Filter Cubes, see page 18
Objective Lenses, see page 20
Fluorescence Imaging
Fluorescence is a property of certain classes of molecules (fluorophores) in which photons of
a specific wavelength are absorbed (excitation), and, as a result, photons are emitted at a
longer wavelength (emission) a very short time later. The utility of fluorescence imaging in
biological applications stems from the ability to conjugate fluorescent molecules with
biologically significant probe molecules, so that visualization of the combined fluorophore in
the specimen highlights the specific distribution of the molecules in question.
By attaching different probes to a set of dye molecules with non-overlapping excitation and
emission spectra, you can stain a specimen with multiple fluorophores and either
simultaneously or sequentially image different structures or substances within the same
specimen. The excitation and emission peaks for each dye or fluorescent protein in a given
environment are physical characteristics of that molecule, and their specific properties
determine the initial selection of the optical components to be used, such as the emission
and excitation filters and the dichroic mirror.

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Filter Cubes
In the ImageXpressPico system, the filter cube contains the excitation and emission filters
and the dichroic mirror. These optical components are essential parts of an epi-illumination
fluorescence imaging system in which the illumination and imaging optical paths overlap at
the objective lens.
A bandpass filter in the illumination optical path (called the excitation filter, since it filters the
excitation light) is used to restrict the illumination spectrum to a narrow range of
wavelengths.
A dichroic mirror is a specially designed beam splitter that transmits light above a certain
cutoff wavelength and reflects light at shorter wavelengths. In the illumination path, the
dichroic mirror reflects shorter wavelengths from the light source up through the objective
onto the specimen.
Similarly, when imaging the illuminated sample, it is desirable to collect only the emission
photons from the target fluorophore, rejecting as much as possible any reflected or
scattered excitation light, any light from other dyes, and autofluorescence from the sample
and substrate. This is done by placing a filter in the collection light path, called the emission
filter, to restrict the wavelength range.
The optics in a filter cube are interference filters made by depositing thin film coatings on a
glass support. These components are delicate and can be easily damaged. Always use care
when handling a filter cube.

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Dichroic Transmission Spectrum
An ideal dichroic mirror would have an infinitely sharp cut-off. That is, it would have unity
transmittance coefficient at wavelengths longer than the cut-off, and zero transmittance
(and therefore unity reflectance in a non-absorbing dichroic mirror) at shorter wavelengths.
In practice, the characteristic transmission spectrum for a dichroic mirror looks similar to the
following graph.
In principle, the cutoff wavelength (or midpoint of the cutoff region) of the dichroic mirror
should be chosen to lie halfway between the absorption and emission peaks of the chosen
fluorochrome, as this simultaneously maximizes the amount of excitation light available at
the sample and also the amount of collected fluorescence emission that is transmitted to the
camera. In practice, however, additional considerations such as fluorochrome efficiency can
dictate that the cutoff region is biased toward one peak or the other. This allows, for
example, greater transmission of longer wavelength image photons at the expense of less
reflection of shorter wavelength excitation light.

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Objective Lenses
The ImageXpressPico system can be configured with high-quality Leica Fluotar objectives.
Five objectives are currently available.
You can identify the magnification of an objective by the color band:
Objective Magnification Color Band
4x Red
10x Yellow
20x Green
40x Light Blue
63x Dark Blue
The 40x objective and 63x objective have application-optimized correction collars (CORR)to
compensate for well bottom thickness or coverslip thickness. The collars have a range of
0mm to 2mm correction. Changing this setting adjusts the distances between components
inside the objective barrel. Image quality and resolution are very dependent on properly
setting these collars.
The settings to be used depend on the well bottom thickness of the plate or the coverslip
thickness on the slide on which the specimen is mounted. In general, set the correction collar
for the physical thickness of the plate or slide that you are imaging. The physical thickness
can be determined by the plate specifications from the plate manufacturer.
See Objectives on page 67 for details.