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'The new EnSight
Multimode Plate Reader’
more predictive results in drug discovery
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© 2013
2009 PerkinElmer
Human
Tissue
Model
organisms
3D
Microtissues
Cells
DNA, RNA,
Protein
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Cost per Data Point
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Image-based Cytometry
Clinical Relevance / System Complexity
Imaging & Detection Translational Research Model
Translating into Clinical Impact
EnSight Multimode Plate Reader with Cell Imaging
Make new findings faster with the only benchtop system
to offer multimode detection, Alpha, label-free & imaging
Get more predictive results with a truly orthogonal approach:
target-based/phenotypic and/or the combination of both
Benefit from the new image-based cytometry module:
4 colors, brightfield/digital phase-contrast imaging at high
speed
(4 min for a single color to read 384-well plate)
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What Kind of Assay Can You Run on The EnSight System?
Cell-based Assays
Cell Imaging
Biochemical Assays
Integrated Well
Integrated Well
Cellular
Sub-cellular
ViewLux® uHTS
Microplate Imager
Throughput
EnVision® Multilabel
Plate Reader
Opera Phenix™
High Content Screening System
EnSight
Multimode Plate Reader
EnSight/
Imaging
Operetta®
High Content Imaging System
VICTOR™ X
Multilabel Plate Reader
Multi-mode Detection World (MMD)
Cytometry
Content
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High Content World (HCS)
EnSight Multimode Plate Reader with Imaging
Software
Kaleido™ Data Acquisition and
Analysis Software
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
Hardware
 New High Value Design
 Add new technologies
 Field upgrades for all
Workflow-based GUI
New Instrument Control &
Data Structure
Customized Data
Evalution
technologies
Cell Imaging
Cell Resolution (e.g. cell counting)
Single
Fast 96/384- well plate reading for 4 colors,
brightfield & digital phase-contrast imaging
Combine with convential MMD
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For Target-Based & Phenotypic Assays
: The new MMD Software
Workflow
Plate map
Measurement Sequence:
Chose your technologies
Content Area:
Results/Well picture
Kaleido for Data Acquisition and Analysis:
 Quickly & easily to set up & run assays using the pre-programmed protocols.
 Analyze & re-analyze your data, and present your results on your office PC to free up the instrument for use.
 Display your results as a list of numerical results, as graphs, in a plate view format or as a heatmap.
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...Is Flexible & Powerful
EnSight Multimode Plate Reader with Imaging
Choose what you need now and be ready for the future
Flexible & modular platform: All technologies are field-upgradeable
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For Target-Based & Phenotypic Assays
Different Technologies for Orthogonal Approaches
Optical Corning® Epic ®
Label-free
Alpha Technology
Image-based Cytometry
Fluorescence Intensity (FI)
http://suzuki-lab.applc.keio.ac.jp/reserches%20English/reserches.html
Absorbance (Abs)
Luminescence (Lumi)
Time-Resolved Fluorescence (TRF)
http://en.wikipedia.org/wiki/Absorbance
http://www.biocompare.com/images/bc/006/ArticleImages/maxline_32_3.jpg
http://media.photobucket.com/image/luminescence%20firefly/Groovy23_Photo
s/Firefly385_359183a.jpg
http://www.case.edu/med/pathology/faculty/cobblab/TRF.html
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One Benchtop Instrument
Camera-Based Widefield Imaging Module
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Key Features
Benefits
3 measurement modes
4 FI excitations, brightfield, digital phase-contrast imaging
Laser-based fast autofocus
Short reading times
High sensitivity camera: scientific CMOS with high S/B ratio
High performance at low light & high speed
Special optical design with no filter movements
Fast & high performance, quick multi-color assays
Useable with all MMD technologies plus label-free
Multiplexing, orthogonal workflow
Designed for Fast & Entire Well Measurements
Stainless Measurement Modes
Stainless (No Label)
(Excitation 740 nm)
Digital Phase-Contrast Imaging
 No labeling
 Non-invasive
 Excellent Signal to Noise Ratio
Brightfield Image
Digital Phase-Contrast (DPI)
 Unique!
 Proprietary algorithms!
2 Slightly Defocused
Brightfield Images
Brightfield
 No labeling
 Non-invasive
 Fast (~4min)
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Algorithm
Digital Phase-Contrast
Image
Digital Phase-Contrast Imaging
Brightfield Image
Digital Phase-Contrast Image
Proprietary Algorithm
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Fluorescence Imaging
Technical details:
4 excitation sources (LEDs):
 UV (LED1: 385 nm), blue (LED2: 470 nm),
green (LED3: 525 nm), red (LED4: 632 nm)
96-well plate: ~1:30 min per single color
384 well plates: ~4 min per single color
Label-based:
Fluorescence Imaging
Fluorescence Intensity
 Specific labeling of specimen
 Label might bias properties or can
be toxic
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Many different labels possible:
Excitation
Dyes (Examples)
385 nm
Hoechst, DAPI
470 nm
FITC, Fluor 3, Calcein, GFP,
AlexaFluor488
525 nm
Texas Red, Bobo3
632 nm
Draq5, Alexa Fluo647
Fluorescence Intensity Mode
HeLa Cells, stained with:
Hoechst (Blue channel/Nuclei)
Overlay
MitoTracker deep red (Green display/Cytoplasm)
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Endless Application Possibilities
Cell counting/
Normalization
Cell Signaling
Cell Toxicity
Phenotypic
Viability
Apoptosis
Proliferation
Cell Migration
Scratch/Oris
Reporter gene
Receptor binding
Cellular Resolution Multiplexing in one Orthogonal
• Endpoint & kinetic Workflow
Confirmation
•
•
•
Fixed & live
•
Label & stainless
Overlay of modes •
Plus conventional •
MMD
•
Plus Label-free
•
Biomarker
Primary & stem
Clone Selection
Transfection rate
Protein expression
Colony forming
Choose what you need now and be prepared for the next project
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Target-based
Phenotypic
Combination
of both
'The new Opera Phenix
High Content Screening
System’
more predictive results in drug discovery
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© 2013
2009 PerkinElmer
Opera Phenix
High Content Screening System
Designed for speed and
sensitivity
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Synchrony™ optics to
deliver and capture more
light of your sample
Simultaneous multi-color
confocal imaging
High Content Screening without compromise
Spectral crosstalk during simultaneous imaging
Opera QEHS
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CROSSTALK
from Hoechst nucleus stain
...can compromise sensitivity
Synchrony Optics™- Proprietary Confocal Lightpath
Transmission LED
1
3
Filter Wheel
Dichro changer
488
488
640
488
375
488Dichro changer
425
405
488
561
488
2
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4
....modular with minimal crosstalk
Synchrony Optics™ – minimal CROSSTALK simultaneous imaging
CROSSTALK
from Hoechst nucleus stain
Opera QEHS
NEW Opera Phenix
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...speed AND sensitivity
High performance objectives
Proprietary high NA water objectives
New supplier: Zeiss
New water collar for greater plate compatibility
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20X water immersion lens, NA 1.0
40X water immersion lens, NA 1.1
63X water immersion lens, NA 1.15
Choice of air objectives
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1.25X air long WD,
5X air long WD,
10X air long WD,
20X air long WD,
40X air long WD,
NA 0.03
NA 0.16
NA 0.3
NA 0.4 (default)
NA 0.6
6 Objective turret


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3 positions for water objectives
3 positions for air objectives
...brighter than ever
Four 4.5 Mpixel sCMOS
SNR sCMOS > CCD at low photon counts
Large field of view – 2.9x Opera
Large dynamic range
Faster frame rates possible
Global shutter for synchronization
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...speed AND sensitivity
Powerful Image Analysis – Made easy
PhenoLOGIC™ – machine learning
 Segmentation and classification as easy as
point and click
Advanced STAR morphology analysis
 Description of cellular phenotypes using
symmetry, compactness and intracellular
distribution of your fluorescence signal
Texture analysis
 Quantification of intensity patterns
Spot
Hole
Ridge
Valley
Intensity
Saddle Edge
Bright
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Operetta
Dark
Made easy
PhenoLOGIC™: supervised machine learning
Train PhenoLOGIC
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Apply segmentation
and classification
Point and Click Segmentation and Classification
NEW Opera Phenix - Benefits for Phenotypic Assays
Confocal Synchrony Optics for effective background reduction
Acquisition of up to 4 channels simultaneously with minimal
Crosstalk for more complete description of phenotypes
Water immersion lenses for higher sensitivity and speed
Large field of view – robust assay statistics
Flexible building blocks for more complete description of
phenotypes
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Opera Phenix - Modular and upgradable
1000
750
500
1 camera
4 laser
2 camera
4 laser
4 camera
5 laser
4 camera
4 xl laser
Basic
Standard
FRET
Screening
250
Operetta
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Meets different needs and price points
PerkinElmer Portfolio for High Throughput Biology
Operetta – the one for everyone


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Exceptional image quality
Powerful Harmony software, yet easy to use
Live cell upgrade
Continuous feature and application development
Opera Phenix – no compromise
 Proprietary Synchrony™ optics to minimize crosstalk
 Powerful Harmony software, yet easy to use
 Truly modular – upgrade from 1 to 2 or 4 confocal cameras
PerkinElmer Total Solution
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Committed to imaging
Spotfire for Your Instruments…
SciStream™ for
Plate Readers
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…stream your scientific data into Spotfire