Molecular Devices ImageXpress Pico User manual

5063666 K
October 2020
ImageXpress Pico
Automated Cell Imaging System
with CellReporterXpress Software Version 2.7
Installation Guide

ImageXpress Pico Automated Cell Imaging System Installation Guide
2 5063666 K
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 do as warranted.
For research use only. Not for use in diagnostic procedures.
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: www.moleculardevices.com/patents
Product manufactured by Molecular Devices, LLC.
3860 N. First Street, San Jose, California, 95134, United States of America.
Molecular Devices, LLC is ISO 9001 registered.
©2020 Molecular Devices, LLC.
All rights reserved.

5063666 K 3
Contents
Safety Information 5
Warnings, Cautions, Notes, and Tips 5
Symbols on Instrument Labels 6
Protective Housing 8
Electrical Safety 8
Moving Parts Safety 11
Lifting Hazard 11
Chemical and Biological Safety 12
Chapter 1: ImageXpressPicoAutomated Cell Imaging System 13
ImageXpressPico System Features 14
CellReporterXpress Software Features 16
Theory of Operation 17
Obtaining Support 20
Product Documentation 21
About This Guide 21
Chapter 2: Setting Up the ImageXpressPico System 23
Package Contents 23
Environmental Requirements 25
Space and Table Requirements 26
Power Requirements 26
Unpacking the Instrument 27
Placing the Instrument 29
Removing the Transport Lock 30
Setting Up the Workstation and Monitor Bundle 30
Connecting Cables 31
Starting the System 33
Chapter 3: Installing the CellReporterXpress Software 35
Chapter 4: Finishing Setup of the ImageXpressPico System 37
Logging In to the Software 37
Registering an Instrument 38
Installing an Objective 41
Installing a Filter Cube 43
Inserting a Labware Holder 45

ImageXpress Pico Automated Cell Imaging System Installation Guide
4 5063666 K
Chapter 5: Environmental Control System 49
Environmental Control System Warnings and Cautions 50
Environmental Control System Hardware 51
Using the Environmental Control System 54
Maintaining the Environmental Control System 64
Troubleshooting the Environmental Control System 64
Appendix A: Instrument Specifications 65
Instrument Dimensions without Environmental Control System 67
Instrument Dimensions with Environmental Control System 68
Appendix B: Optional Parts and Accessories 71
Objectives 71
Filter Cubes 71
Accessories 72
Appendix C: Environmental Control System Gas Regulator Details 73
Using a Gas Cylinder 74
Using an Oil-Free Lab Gas Line 76
Using an Oil-Free Air Compressor 77
Unsupported Gas Supplies 77
Appendix D: Packing the Instrument for Transport 79
Appendix E: Electromagnetic Compatibility 87

5063666 K 5
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.

ImageXpress Pico Automated Cell Imaging System Installation Guide
6 5063666 K
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. To prevent injury, use a minimum of two people to
lift the instrument. See Lifting Hazard on page 11 for details on the weight of the
instrument.
Indicates a potential pinch hazard.
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
5063666 K 7
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.

ImageXpress Pico Automated Cell Imaging System Installation Guide
8 5063666 K
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 top 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.
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.
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
5063666 K 9
Replacing Fuses
If the instrument does not power on when 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 fails 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 65 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.

ImageXpress Pico Automated Cell Imaging System Installation Guide
10 5063666 K
To replace fuses:
1. If the instrument is powered on, press the Power button on the front of the instrument to
turn off the power.
When the status light on the Power button is amber, the instrument is fully powered off.
Note: If the ImageXpress Pico system doesn't respond, you can perform a hard
shutdown by holding the Power button for five seconds.
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.
When the light on the Power button is green, the instrument is fully powered on.
Note: If the instrument still does not power on after replacing fuses, contact Molecular
Devices Technical Support. See Obtaining Support on page 20 for details.

Safety Information
5063666 K 11
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.

ImageXpress Pico Automated Cell Imaging System Installation Guide
12 5063666 K
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 an instrument that is powered on.
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.

5063666 K 13
Chapter 1: ImageXpress Pico Automated Cell Imaging
System
The ImageXpress® Pico Automated Cell Imaging System is an 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.
1

ImageXpress Pico Automated Cell Imaging System Installation Guide
14 5063666 K
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 71 for details.
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.
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
4x 0.13
10x 0.32
20x 0.40
40x 0.60
63x 0.70
CAUTION! To prevent damaging both the instrument and your sample, do not use any
other objectives with the ImageXpress Pico system.
See Objectives on page 71 for details.
Motorized Objective Turret
The ImageXpress Pico system includes a six-position objective turret.

Chapter 1: ImageXpressPicoAutomated Cell Imaging System
5063666 K 15
Sample (X-Y) Stage
Plate Holder
The plate holder is designed for imaging multi-well microplates in standard ANSI (SBS) formats
with plastic or glass bottoms and standard footprint dimensions. It can also be used with the
optional dish holder, which holds up to six 35mm (1.38in.) dishes. Optimal image quality
depends on plate flatness, well bottom thickness, and optical clarity.
A spring-loaded, mechanical clamp holds the plate or dish holder 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 into position for imaging.
Slide Holder
The slide holder is designed for imaging up to four standard microscope slides of 25mm x
75mm (1in. x 3in.).

ImageXpress Pico Automated Cell Imaging System Installation Guide
16 5063666 K
Dish Holder
The optional dish holder is used with the plate holder. It is designed for imaging up to six
standard culture dishes of 35mm (1.38in.) in diameter.
CellReporterXpress Software Features
The CellReporterXpress Image Acquisition and Analysis Software 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.
Note: See the CellReporterXpress Help or the CellReporterXpress User Guide for
details on using the CellReporterXpress software.
If you purchased the optional workstation and monitor bundle, the CellReporterXpress
software is pre-installed on the host computer at the time of delivery. You may still need to
install the software on any remote analysis and remote storage computers. You may also want
to set up workstation clients.
If you did not purchase the optional workstation and monitor bundle, you will install the
CellReporterXpress software as part of setting up the ImageXpress Pico system.

Chapter 1: ImageXpressPicoAutomated Cell Imaging System
5063666 K 17
Theory of Operation
The ImageXpress Pico system uses the following components and functions:
Fluorescence Imaging, see below
Objective Lenses, see page 18
Filter Cubes, see page 19
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 use, such as the emission and
excitation filters and the dichroic mirror.

ImageXpress Pico Automated Cell Imaging System Installation Guide
18 5063666 K
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 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 71 for details.

Chapter 1: ImageXpressPicoAutomated Cell Imaging System
5063666 K 19
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.

ImageXpress Pico Automated Cell Imaging System Installation Guide
20 5063666 K
Obtaining Support
Molecular Devices is a leading worldwide manufacturer and distributor of analytical
instrumentation, software, and reagents. We are committed to the quality of our products and
to fully supporting our customers with the highest level of technical service.
Our Support website, www.moleculardevices.com/service-support, describes the support
options offered by Molecular Devices, including service plans and professional services. It also
has a link to the Molecular Devices Knowledge Base, which contains documentation, technical
notes, software upgrades, safety data sheets, and other resources. If you still need assistance,
you can submit a request to Molecular Devices Technical Support.
Technical Support
You can contact Molecular Devices Technical Support by submitting a support request through
the Molecular Devices Knowledge Base at mdc.custhelp.com.
You can also submit a support request by phone. For regional support contact information, go
to www.moleculardevices.com/contact.
You will need the instrument serial number. The serial number is located on the back panel of
the instrument.