Operetta The Next Generation in High Content Screening

Operetta
Compact High Content Screening Reader
the next generation
in high content
screening

Operetta
Operetta –
the high content
screening solution
for every lab
Offering a new bench-top
approach to High Content
Screening (HCS), the
Operetta is easy to use,
allowing researchers who are new to HCS to use preconfigured
applications and quickly become productive. User-configurable
building blocks enable more experienced users to develop their
own assays, or modify existing applications. The compact design
fits any size laboratory to provide a versatile solution that suits
the needs of every HCS facility.
Benefiting from many years of experience creating outstanding
solutions for HCS, PerkinElmer brings new, state-of-the-art
technology to HCS research. Our in-depth knowledge of the
technical requirements for successful high content imaging is
complemented by our understanding of researchers' assays and
applications. Providing a total solution for beginners and experts
alike, the Operetta incorporates both engineering excellence and
a complete, easy-to-use software interface.
Key Features
• Easy-to-use solution for High Content Screening –
intuitive user interface, short learning curve and
online user guides enable fast results
• Exceptional image quality
• Powerful image analysis includes automatic parameter
optimization for ease-of-use
• Database with keyword browser enables searching
by user, application, date, screen and compound
Operetta Application Guide
To help new HCS users, we provide an easy to follow
guide to analyzing your results. This step-by-step
handbook will help you through the process, whether
you are using a ready-made solution or designing a
new image analysis sequence from the building blocks
provided. The guide includes general considerations,
recommendations and common pitfalls, so that
you can proceed with confidence and produce
results quickly.
The Operetta has a compact design that will fit into any laboratory.
The large working area beneath the lid allows easy access to samples.
2

Operetta Hardware
Designed to provide the best functionality in the smallest
possible footprint, the Operetta has a sleek profile and status
indicator illumination that allows you to track the progress
of your experiment with ease. Lifting the lid on the Operetta
reveals a large working area allowing easy access to the sample
area, while the eight position emission filter wheel is conveniently
housed so that filters can be changed quickly and easily.
Barcoding prevents filter set-up errors, and maximum flexibility
is achieved with up to 15 emission filter combinations.
All standard objectives are barcoded for convenience, providing
a wide range of choices for any plate type. High numerical
apertures enable optimal resolution and sensitivity. Long working
distance objectives allow the use of any plate type. The
custom-designed light path permits a large field of view, greatly
increasing the number of cells visualized without compromising
resolution and throughput.
Available with brightfield, widefield, and confocal options, the
system can cater for your application needs both now and in
the future. High resolution is provided with a Nipkow disk to
enhance axial resolution and to suppress out of focus light.
Xenon
lamp
Transmission light
Attenuator
Sample
Excitation filter
wheel
Objective turret
Shutter
Shutter
Emission filter
wheel
Camera
Dichroic mirror
Optional confocal light
path, pinhole disk
Autofocus
The optical path of the Operetta
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
Live / Dead Cell Counting
Quantification of Marker in Nucleus
Quantification of Marker in Cytoplasm
Quantification of Marker in Membrane
Cytosol to Nucleus Translocation
Cytosol to Membrane Translocation
Fluorescence Redistribution – Cytoskeleton
proliferation, cytotoxicity
cell viability: live / dead cell count
Protein expression in nucleus (e.g. caspase-3) • Cell cycle: S-Phase quantification (BrdU, EdU)
Protein expression in cytosol • Mitochondrial mass • Loss of mitochondrial membrane potential
Phospholipidosis • Hepatosteatosis • Cell cycle: Transition of G2 to M-phase (cyclin B1)
Protein expression in plasma membrane • Signalling pathways: Membrane ruffling
Signalling pathways: Transcription factor activation
Signalling pathways: Cytoplasm to membrane translocation
Cytoskeletal rearrangement
Spot Analysis Peroxisome quantification • Vesicle formation • GPCR activation: arrestin recruitment (e.g. Transfluor ® )
Nuclear Spots
Nuclear Analysis – Nuclear Shrinking
Nuclear Fragmentation
Nuclear Classification – DNA Content
Cell Shape – Cell Rounding
fluorescence In Situ Hybridization (FISH)
Nuclear swelling and shrinking
apoptosis / Nuclear fragmentation
Cell cycle: DNA content
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.
4

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
Find Cytoplasm
Find Cells
Find Spots
Find Image Region
Select Cell Region
Select Region
Calculate Intensity Properties
Calculate Morphology Properties
Calculate Properties
Select Population
identifies and counts cellular nuclei based on a nuclear stain
Identifies cell bodies / cytoplasmic area when nuclei have been identified before
Finds whole cells based on fluorescence intensity e.g. for whole cell fluorescence assays
Identifies fluorescent objects in cells of similar size, such as endosomes, vesicles, peroxisomes
Segments the entire image based on fluorescence intensity
Defines a region in the cytoplasm or nucleus, based on nucleus and cytoplasm identification
Defines a sub-region
calculates fluorescence intensity of regions / objects, provides mean,
median, standard deviation, CV, max, min, contrast values
Calculates morphology properties of regions / objects, such as area, length, width,
axis, ratio, roundness
Allows any combination of intensity and morphology properties
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
Light sources
• Xenon fibre optic light source, 300 W,
360 – 640 nm continuous spectrum
• LED light source for transmission mode
Excitation and emission
• Fully automated eight position excitation filter wheel
• Fully automated eight position emission filter wheel
• 15 emission filters available, barcoded and user exchangeable
Camera
• 14-bit Peltier-cooled camera
• CCD chip size 1360 x 1024 pixel, 6.45 µm square
Objectives
• 4 position objective turret for automated change
• 10x high NA and long WD
• 20x high NA and long WD
• 40x long WD
• 60x long WD
• 100x long WD
Scan table
• High resolution scanning stage
(150 nm mechanical stage x-y accuracy)
Harmony Software
Image acquisition
• Computer controlled excitation power
• Laser-based fully automated and fast autofocusing
• Preconfigured plate formats (96, 384)
• User defined plate formats; free definition also possible
• Assay layout creation with arbitrary layers of information
• Multiple image fields within wells
• Z-stack measurements
• Fully network compatible
Image analysis
• Fully automated cell segmentation with automatic
or manual parameter tuning
• Ready-made turnkey applications
• Easy and intuitive creation and modification of analysis
sequences
Data management
• Scalable database
• Search via sortable tree
• Stores all metadata automatically
(assay layout, instrument setting, analysis results)
• Stores additional experiment information
(user defined keywords, annotations)
• Allows for image storage on a remote server
Computer
Processor
Memory
Hard Drive
Network interface
Intel ® Core 2 Quad 2,83 GHz
4 GB
500 GB (RAID 1, 2 x 500 GB)
Operating system Windows Vista ®
Gigabit Ethernet
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.
6

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
Set up on the Operetta system
or transfer to the Operetta
from your office computer via
the network when required.
Optional steps
Develop/modify image analysis,
re-analyze images
Select measurement
protocol
Perform on the Operetta system, or
transfer to your office computer via
the network.
Select image analysis
solution
Automated measurement
Online image analysis
View results
View on the Operetta system,
or transfer to your office
computer via the network.
www.perkinelmer.com/operetta
7

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
Screening System
The Opera FLIM is a time-domain
fluorescence lifetime imaging reader
with ultra-high speed data acquisition.
Opera QEHS High Content
Screening System
The Opera is the premier confocal
microplate imaging reader providing
solutions for fully automated simultaneous
multi-color high speed and high resolution
screening.
Acapella Image Analysis Software
Acapella is a flexible and powerful
image data processing system for a
broad range of applications.
Opera LX High Content
Screening System
The Opera LX confocal microplate
imaging reader is an extendable starter
configuration for three color applications.
Operetta Compact High
Content Screening System
The Operetta is an easy-to-use
bench-top high content screening
instrument. Includes an intuitive
user interface, multiple wavelength
fluorescence and brightfield imaging.
Columbus Data Management
and Analysis Software
Columbus sets the industry standard for
high volume image data management
providing image storage, retrieval,
visualization and analysis.
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