Operetta The Next Generation in High Content Screening

the next generation

in high content

screening

Operetta

Compact High Content Screening Reader

Operetta

Operetta –
the high content

screening solution Offering a new bench-top Key Features
approach to High Content • Easy-to-use solution for High Content Screening –
for every lab Screening (HCS), the
intuitive user interface, short learning curve and
Operetta™ is easy to use, online user guides enable fast results
allowing researchers who are new to HCS to use preconfigured • Exceptional image quality
applications and quickly become productive. User-configurable • Powerful image analysis includes automatic parameter
building blocks enable more experienced users to develop their optimization for ease-of-use
• Database with keyword browser enables searching
own assays, or modify existing applications. The compact design by user, application, date, screen and compound

fits any size laboratory to provide a versatile solution that suits Operetta Application Guide
the needs of every HCS facility. To help new HCS users, we provide an easy to follow
guide to analyzing your results. This step-by-step
Benefiting from many years of experience creating outstanding handbook will help you through the process, whether
solutions for HCS, PerkinElmer brings new, state-of-the-art you are using a ready-made solution or designing a
technology to HCS research. Our in-depth knowledge of the new image analysis sequence from the building blocks
technical requirements for successful high content imaging is provided. The guide includes general considerations,
complemented by our understanding of researchers' assays and recommendations and common pitfalls, so that
applications. Providing a total solution for beginners and experts you can proceed with confidence and produce
alike, the Operetta incorporates both engineering excellence and results quickly.
a complete, easy-to-use software interface.

The Operetta has a compact design that will fit into any laboratory.
The large working area beneath the lid allows easy access to samples.
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Operetta Hardware All standard objectives are barcoded for convenience, providing
a wide range of choices for any plate type. High numerical
Designed to provide the best functionality in the smallest apertures enable optimal resolution and sensitivity. Long working
possible footprint, the Operetta has a sleek profile and status distance objectives allow the use of any plate type. The
indicator illumination that allows you to track the progress custom-designed light path permits a large field of view, greatly
of your experiment with ease. Lifting the lid on the Operetta increasing the number of cells visualized without compromising
reveals a large working area allowing easy access to the sample resolution and throughput.
area, while the eight position emission filter wheel is conveniently
housed so that filters can be changed quickly and easily. Available with brightfield, widefield, and confocal options, the
Barcoding prevents filter set-up errors, and maximum flexibility system can cater for your application needs both now and in
is achieved with up to 15 emission filter combinations. the future. High resolution is provided with a Nipkow disk to
enhance axial resolution and to suppress out of focus light.

Xenon Transmission light
lamp Sample
Objective turret
Attenuator
Shutter
Excitation filter Shutter
wheel

Emission filter
wheel

Camera Dichroic mirror

The optical path of the Operetta Optional confocal light Autofocus
path, pinhole disk

www.perkinelmer.com/operetta 3

Operetta – hardware and software in complete harmony

Providing shared, remote and dedicated access, the Harmony™ software is designed to streamline
and simplify the HCS process. With its intuitive user interface that leads you through your
HCS experiment, Harmony suggests next steps where appropriate, simplifying the process
for both new and experienced HCS users.

Harmony includes all the tools you need to develop an assay, automate the hardware, analyze your data, and store and retrieve
the results. Remote access to the Operetta is available via your office computer and local network for analyzing and viewing
your results.

Harmony for Flexible Acquisition

Acquisition controls are displayed within a unified window with predefined solutions or options to create protocols to your own
requirements. Multiple acquisition options are available, including channels, fields per well and z-sections. All imaging parameter
metadata, such as dyes / filters, objectives, plate type, fields, z-stack or exposure, are embedded into the experiment files and
stored in the database for future reference.

Harmony for Rapid, Accurate Analysis
Ready-Made Solutions

A full range of ready-made solutions for frequently used image analysis tasks and assays is pre-installed for your convenience,
making it ideal for new users. Harmony still provides the flexibility to change and modify your experiments using image
analysis building blocks.

Harmony Ready-Made Solution Applications (examples)

Cell Counting or Nuclei Counting Proliferation, cytotoxicity

Live / Dead Cell Counting Cell viability: live / dead cell count

Quantification of Marker in Nucleus Protein expression in nucleus (e.g. caspase-3) • Cell cycle: S-Phase quantification (BrdU, EdU)

Quantification of Marker in Cytoplasm Protein expression in cytosol • Mitochondrial mass • Loss of mitochondrial membrane potential
Phospholipidosis • Hepatosteatosis • Cell cycle: Transition of G2 to M-phase (cyclin B1)

Quantification of Marker in Membrane Protein expression in plasma membrane • Signalling pathways: Membrane ruffling

Cytosol to Nucleus Translocation Signalling pathways: Transcription factor activation

Cytosol to Membrane Translocation Signalling pathways: Cytoplasm to membrane translocation

Fluorescence Redistribution – Cytoskeleton Cytoskeletal rearrangement

Spot Analysis Peroxisome quantification • Vesicle formation • GPCR activation: arrestin recruitment (e.g. Transfluor®)

Nuclear Spots Fluorescence In Situ Hybridization (FISH)

Nuclear Analysis – Nuclear Shrinking Nuclear swelling and shrinking

Nuclear Fragmentation Apoptosis / Nuclear fragmentation

Nuclear Classification – DNA Content Cell cycle: DNA content

Cell Shape – Cell Rounding Cell spreading, cell detachment, cell rounding

Mitotic Index Cell cycle: Mitotic index (phosphohistone H3)

Cell Cycle Classification Simultaneous phosphohistone H3, EdU, DNA content and nuclear morphology

Harmony includes preconfigured image analysis sequences for common applications. Known as ready-made solutions, they are ideal for new users of HCS.

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Custom-Designed Solutions

Harmony provides a complete range of building blocks to enable simplified custom image analysis. Each clearly defined block has
primary tuning parameters with optimized values that can be automatically or manually adjusted with real-time visual feedback to
facilitate the set-up process.

Harmony Building Block Description

Find Nuclei Identifies and counts cellular nuclei based on a nuclear stain

Find Cytoplasm Identifies cell bodies / cytoplasmic area when nuclei have been identified before

Find Cells Finds whole cells based on fluorescence intensity e.g. for whole cell fluorescence assays

Find Spots Identifies fluorescent objects in cells of similar size, such as endosomes, vesicles, peroxisomes

Find Image Region Segments the entire image based on fluorescence intensity

Select Cell Region Defines a region in the cytoplasm or nucleus, based on nucleus and cytoplasm identification

Select Region Defines a sub-region

Calculate Intensity Properties Calculates fluorescence intensity of regions / objects, provides mean,
median, standard deviation, CV, max, min, contrast values

Calculate Morphology Properties Calculates morphology properties of regions / objects, such as area, length, width,
axis, ratio, roundness

Calculate Properties Allows any combination of intensity and morphology properties

Select Population Defines sub-populations of cells

Harmony for Data Storage and Retrieval

A scalable database is included for management of all images, experiment information and data analysis. Metadata information,
such as exposure parameters, analysis method and users, is embedded into all data files, while a keyword browser facilitates
searching using those keywords for your projects. A sortable database information tree allows quick and easy access to any
desired information.

Automatic parameter adjustment helps you find the best settings quickly. Find images, metadata and results easily in the Harmony database.

www.perkinelmer.com/operetta 5

Operetta — compact instrument providing the complete solution

Operetta Hardware Harmony Software
Light sources Image acquisition
• Xenon fibre optic light source, 300 W, • Computer controlled excitation power
• Laser-based fully automated and fast autofocusing
360 – 640 nm continuous spectrum • Preconfigured plate formats (96, 384)
• LED light source for transmission mode • User defined plate formats; free definition also possible
• Assay layout creation with arbitrary layers of information
Excitation and emission • Multiple image fields within wells
• Fully automated eight position excitation filter wheel • Z-stack measurements
• Fully automated eight position emission filter wheel
• 15 emission filters available, barcoded and user exchangeable • Fully network compatible

Camera Image analysis
• 14-bit Peltier-cooled camera • Fully automated cell segmentation with automatic
• CCD chip size 1360 x 1024 pixel, 6.45 µm square
or manual parameter tuning
Objectives • Ready-made turnkey applications
• 4 position objective turret for automated change • Easy and intuitive creation and modification of analysis
• 10x high NA and long WD
• 20x high NA and long WD sequences
• 40x long WD
• 60x long WD Data management
• 100x long WD • Scalable database
• Search via sortable tree
Scan table • Stores all metadata automatically

• High resolution scanning stage (assay layout, instrument setting, analysis results)
(150 nm mechanical stage x-y accuracy) • Stores additional experiment information

(user defined keywords, annotations)

• Allows for image storage on a remote server

Computer Intel® Core™ 2 Quad 2,83 GHz
Processor 4 GB
Memory 500 GB (RAID 1, 2 x 500 GB)
Hard Drive Operating system Windows Vista®
Gigabit Ethernet
Network interface
24" flat screen


Physical Dimensions

Width: 33" (83 cm)
Depth: 21" (53 cm)
Height: 16" (40 cm)

Operetta status indicator illumination helps monitor
the progress of experiments.

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Operetta – use the network solution to maximize results

By using the local network, you can perform tasks such as experiment design and viewing
results, so that the Operetta is free for other researchers to use, maximizing the productivity
of your system.

Define assay layout Optional steps
Set up on the Operetta system Develop/modify image analysis,
or transfer to the Operetta re-analyze images
from your office computer via Perform on the Operetta system, or
the network when required. transfer to your office computer via
the network.
Select measurement
protocol View results
View on the Operetta system,
Select image analysis or transfer to your office
solution computer via the network.

Automated measurement
Online image analysis

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PerkinElmer support to suit your needs

PerkinElmer is committed to providing outstanding after-sales support using our network of local, highly trained service engineers. As
an Operetta user you can choose the level of support that you require, from maintenance agreements for hardware or software, right
up to full 24 hour response.

PerkinElmer’s High Content Screening product solutions

Opera FLIM High Content Opera QEHS High Content Acapella Image Analysis Software
Screening System Screening System
Acapella™ is a flexible and powerful
The Opera™ FLIM is a time-domain The Opera™ is the premier confocal image data processing system for a
fluorescence lifetime imaging reader microplate imaging reader providing broad range of applications.
with ultra-high speed data acquisition. solutions for fully automated simultaneous
multi-color high speed and high resolution
screening.

Opera LX High Content Operetta Compact High Columbus Data Management
Screening System Content Screening System and Analysis Software

The Opera™ LX confocal microplate The Operetta™ is an easy-to-use Columbus™ sets the industry standard for
imaging reader is an extendable starter bench-top high content screening high volume image data management
configuration for three color applications. instrument. Includes an intuitive providing image storage, retrieval,
user interface, multiple wavelength visualization and analysis.
fluorescence and brightfield imaging.

Disclaimer: A user license to the ThermoFisher Scientific (Cellomics) HCS patent portfolio is included with each Operetta™ and Harmony™ product.
For the use of the assays described additional licenses may be required.

PerkinElmer, Inc.
940 Winter Street
Waltham, MA 02451 USA
P: (800) 762-4000 or
(+1) 203-925-4602
www.perkinelmer.com

For a complete listing of our global offices, visit www.perkinelmer.com/ContactUs

Copyright ©2009, PerkinElmer, Inc. All rights reserved. PerkinElmer® is a registered trademark of PerkinElmer, Inc. All other trademarks are the property of their respective owners.

008451B_01 Printed in USA