Molecular Devices ImageXpress Pico User manual

5063666 B
June 2018
ImageXpress®Pico
Automated Cell Imaging System
Installation Guide

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Contents
Safety Information 5
Warnings, Cautions, Notes, and Tips 5
Symbols on Instrument Labels 6
Before Operating the Instrument 7
Protective Housing 7
Electrical Safety 8
Moving Parts Safety 9
Lifting Hazard 10
Chemical and Biological Safety 10
Cleaning and Maintenance 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 20
Product Documentation 22
About This Guide 22
Chapter 2: Setting Up the ImageXpressPico System 23
Package Contents 23
Unpacking the Instrument 24
Placing the Instrument 26
Removing the Transport Lock 26
Connecting Cables 27
Powering On the Instrument 29
Chapter 3: Installing the CellReporterXpress Software 31
Chapter 4: Finishing Setup of the ImageXpressPico System 33
Logging In to the Software 33
Installing a Filter Cube 34
Installing an Objective 36
Inserting the Plate Holder or Slide Holder 38

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Appendix A: Instrument Specifications 41
Instrument Dimensions 43
Appendix B: Compatible Objectives 47
Appendix C: Filter Cube Specifications 49
Appendix D: Replacement Parts and Optional Extras 51
Appendix E: Packing the Instrument for Transport 53
Appendix F: Electromagnetic Compatibility 57

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Safety Information
The safety information section provides information on the safe use of the instrument. It
includes the use of user-attention statements in this guide, a key to understanding the
safety labels on the instrument, precautions to follow before operating the instrument, and
precautions to follow while operating the instrument.
Read and observe all warnings, cautions, and instructions. Remember, the most important
key to safety is to operate 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.
This symbol on the product is required in accordance with the Waste Electrical and
Electronic Equipment (WEEE) Directive of the European Union. It indicates that 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.
Indicates that there is an authorized representative in the European community.
Indicates the instrument manufacturer.
Indicates the manufacturer catalog number.
Before Operating the Instrument
Make sure 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.
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 20 cm to 30 cm (7.9 in. to 11.8 in.) gap between the rear of the
instrument and the wall.
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 8 for details.
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. You can obtain
replacement fuses from Molecular Devices. See Replacement Parts and Optional Extras on
page 51 for details on fuse specifications and part numbers.
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 part of the power outlet on the rear of the
instrument.

Safety Information
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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. Use a small flat-head screwdriver to gently press on the carrier-release tab and then 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 20 for details.
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.

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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! Moving the instrument can damage sensitize 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.
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. Therefore, take applicable safety precautions, such as
using safety glasses and wearing protective clothing, when working with potentially
hazardous liquids.
Observe the applicable cautionary procedures as defined by your safety officer when
using hazardous materials.
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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Cleaning and Maintenance
Observe the cleaning procedures outlined in this guide for the instrument.
Do the following before you clean equipment that has been exposed to hazardous material:
Contact the applicable Chemical and Biological Safety personnel.
Review the Chemical and Biological Safety information contained in this guide. See
Chemical and Biological Safety on page 10 for details.
Perform only the maintenance tasks described in this guide. Any other maintenance tasks
must be done by qualified Molecular Devices personnel only. Contact a Molecular Devices
service engineer to inspect and perform a preventive maintenance service on the instrument
each year.
WARNING! BIOHAZARD. It is your responsibility to decontaminate
components of the instrument before you return parts to Molecular Devices for
repair. Molecular Devices does not accept items that have not been
decontaminated where it is applicable to do so. If parts are returned, they must be
enclosed in a sealed plastic bag stating that the contents are safe to handle and
are not contaminated.
For approved cleaning and maintenance procedures, see the ImageXpress Pico User Guide.

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Chapter 1: ImageXpressPicoAutomated Cell Imaging
System
The ImageXpress®PicoAutomated Cell Imaging System by Molecular Devices® 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.
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, with asynchronous command execution, allowing 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 over 25 available
predefined experimental protocols, the ImageXpressPico System becomes an extremely
flexible and programmable device, ideally suited for user-defined, high-speed automated
assays.
Key components of the instrument include the following:
Built-in, internal light source comprised of four high-powered LEDs enables very high
sensitivity fluorescent imaging. Additional LED-based light sources allow transmitted light
and planetary view 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.
Planetary view 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 for live cell imaging at 6°C (11°F) above ambient up to 40°C (104° F).
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 four 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 Cube Specifications on page 49 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 Compatible Objectives on page 47 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 microplate footprint dimensions. For example, glass slides can be imaged using a
slide adapter included in the accessory kit. 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 (X-Y) stage moves to the load/eject position; it
automatically closes when the sample (X-Y) 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 higher
magnifications, you can select an additional image-based autofocus.

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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 imaging device, the CellReporterXpress Software is part of a system
that 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 installing and setting up the ImageXpressPico System, you will install the
CellReporterXpress Software on a host computer.
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 page 18
Excitation and Emission Filters, see page 18
Dichroic Mirror, see page 19
Objective Lenses, see page 20

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Fluorescence Imaging
Fluorescence is a property of certain classes of molecules (fluorochromes, fluorescent
proteins, or dyes) 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 active probe molecules, so that application
of the combined dye/probe molecule (fluorophore) to the specimen highlights the specific
substances or regions to which the probe is targeted.
By attaching different probes to a set of dye molecules with non-overlapping excitation and
emission spectra, one can stain a specimen with multiple fluorophores, and either
simultaneously or sequentially image different structures or substances within the same
specimen. The absorption 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.
Excitation and Emission Filters
In the ImageXpressPico System, the excitation and emission filters are located in a filter
cube.
To selectively excite one fluorophore more intensely than another, or to minimize excitation
channel crosstalk, it is necessary to provide illumination containing only photons with a
wavelength range matched to the absorbance (excitation) spectrum of the target dye. 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.
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. Emission filters can either be a bandpass filter (for maximum specificity) or a longpass
filter (to maximize the amount of emission light collected).

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Dichroic Mirror
In the ImageXpressPico System, the dichroic mirror is in a filter cube.
A dichroic mirror is a specially designed beam splitter that transmits light above a certain
cutoff wavelength and reflects light at shorter wavelengths. This is the essential component
that allows the construction of an epi-illumination fluorescence imaging system in which the
illumination and imaging optical paths overlap at the objective lens. The same objective lens
is used to focus the illumination light on the sample as well as collect the emitted fluorescent
light to form the image.
In the illumination path, the dichroic mirror reflects shorter wavelengths from the light
source up through the objective onto the specimen.
In the imaging optical path, longer wavelength fluorescence light emitted by the excited
fluorophores in the specimen is collected by the objective lens and transmitted through the
dichroic mirror to the camera. Incident light from the sample that is shorter wavelength than
the cutoff (mostly reflected illumination light from the sample) is reflected by the dichroic
mirror (and further blocked by the emission filter), preventing it from entering the imaging
system of tube lens and camera.
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.
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
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 external influences such as 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 Compatible Objectives on page 47 for details.
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/support, has a link to the Knowledge
Base with technical notes, software upgrades, safety data sheets, and other resources. If you
still need assistance, submit a request to Molecular Devices Technical Support.
Documentation
Review the product documentation on the Knowledge Base, including installation guides and
user guides. In addition, online Help is available within the CellReporterXpress Software.
Press F1 to access Help for the active screen.
Technical Support
You can contact Molecular Devices Technical Support by phone or submit a support request
through the Knowledge Base. To find regional support contact information, visit
www.moleculardevices.com/contact.
You will need the instrument serial number and the software system ID.