erzeugung und klassifizierung größerer datenmengen im high ... · high-content screening...
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Erzeugung und Klassifizierung größerer
Datenmengen im High-Content ScreeningOlympus scanR
Product Unit Life Science Research | Olympus Soft Imaging Solutions | Dr. Mike Wördemann
Linz, 28th Nov. 2017
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High-Content Screening
▪ Automated, image-based cytometry
▪ Screening at cellular level
▪ Simultaneous multi-parameter readout
▪ Demands
• Robustness
• Reliability
• Automation
• Quantification
▪ For biological research and drug discovery
Olympus Soft Imaging SolutionsPage 2
https://www.olympus-lifescience.com
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Statistics Volume Comparison
Olympus Soft Imaging SolutionsPage 3
Imaging
System
10
9
Flow Cytometer
Plate Reader
Sample throughput
Cells
/ s
am
ple
10
61
03
100.0001000
Image-based
Cytometry
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Workflow
Olympus Soft Imaging SolutionsPage 4
Processing & Analysis
(image level)Data Analysis Result OutputAcquisition
Automated
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High Throughput
Multi well micro plates, for example 96W, 384W
Automatic well plate loader
Full automation, integration into sample preparation pipelines
Olympus Soft Imaging SolutionsPage 5
Source: Olympus marketing video
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Cytometric data exploration
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Example: Multi colour beads
– Image acquisition
– Image processing
– Object segmentation
– Parameter extraction
Olympus Soft Imaging Solutions
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Cytometric data exploration
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Example: Multi colour beads
Exploring data to find the relevant parameters
Olympus Soft Imaging Solutions
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Cytometric classification
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Example: Multi colour beads
Classification in
parameter space
Olympus Soft Imaging Solutions
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Cytometric classification
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Example: Multi colour beads
Cascaded „Gating“ enables even
complex classification tasks
Olympus Soft Imaging Solutions
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Cytometric classification
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Example: Mitotic analysis of fixed cells (ensemble-based)
2x DNA content
1x DNA content
Olympus Soft Imaging Solutions
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Cytometric classification
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Example: Mitotic analysis of fixed cells (ensemble-based)
2x DNA content
1x DNA content
Mitotic phase (~1%)
micro well plate distribution
Olympus Soft Imaging Solutions
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Life cell cytometry
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Example: Mitotic analysis of life cells (time-based)
Cytometric concept can be transfered to time-varying data
Extract kinetic parameters from curves of static parameters, for example:
– GFP mitosis marker over time peaks during mitosis
– Static parameter: GFP content
– Kinetic parameter
1. maximum peak value of GFP over time
2. average GFP value over time
– classification on kinetic parameters
Olympus Soft Imaging Solutions
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Life cell cytometry
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Example: Mitotic analysis of life cells (time-based)
Cytometric concept can be transfered to time-varying data
Extract kinetic parameters from curves of static parameters, for example:
– GFP mitosis marker over time peaks during mitosis
– Static parameter: GFP content
– Kinetic parameter
1. maximum peak value of GFP over time
2. average GFP value over time
– classification on kinetic parameters
micro well plate distribution
Olympus Soft Imaging Solutions
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Conclusions
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Cytometric classification concept allows intuitive classification based on biological
properties (phenotype and labels)
“Rare events” can easily be identified
Can be extened to time-varying data
Scales well up to many 10 million data points
Challenges and potential future Requirements include label-free assays, faster data
generation (more statistics), and ML-based image segmentation
Olympus Soft Imaging Solutions
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