Technology Integration for Analysis of High Content/Throughput Cellular Data: The Cytomics Approach J. Paul Robinson Professor of Immunopharmacology & Professor of Biomedical Engineering Purdue University This presentation will discuss current ideas for analysis of live cell data incorporating multivariate approaches. It will outline the major problems faced by present generation technologies and provide insight into future advances. Key to the success of future technologies will be an understanding of informatics and high-speed data processing including advanced image analysis. www.cyto.purdue.edu
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J. Paul Robinson Professor of Immunopharmacology & Professor of Biomedical Engineering
Technology Integration for Analysis of High Content/Throughput Cellular Data: The Cytomics Approach. J. Paul Robinson Professor of Immunopharmacology & Professor of Biomedical Engineering Purdue University - PowerPoint PPT Presentation
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Technology Integration for Analysis of High Content/Throughput Cellular Data: The Cytomics Approach
J. Paul RobinsonProfessor of Immunopharmacology &Professor of Biomedical EngineeringPurdue University
This presentation will discuss current ideas for analysis of live cell data incorporating multivariate approaches. It will outline the major problems faced by present generation technologies and provide insight into future advances. Key to the success of future technologies will be an understanding of informatics and high-speed data processing including advanced image analysis.
www.cyto.purdue.edu
Goals of this Presentation
Introduce Cytomics Identify current & forthcoming
issues & technologies Call attention to issues that need to
be addressed
Note: Added for the web version of this presentation:
These slides were perfect!! In Powerpoint. However, as amazing as it might seem, the Powerpoint web converter is pretty much the usual Microsoft Disaster Product (MDP).
So, many animations and boxes with text look strange and the text often fails to remain inside the boxes, lines are everwhere….
What is the Cytomics Approach? Discovering the functional relationships between the
cell (Cytome) and the metabolic pathways (Proteomics-proteome) resulting from genetic control mechanisms (Genomics-genome) –
Some relate Cytomics to what is being termed functional genomics.
By definition we are expanding the information being collected in every system because we also want functional data, not just morphological, phenotypic or genotypic.
..the cell is the ultimate functional endpoint…
Cytomics is going to be important because it is the cell that is the ultimate functional endpoint. The cell is the minimal functional unit within our physiology and thus the functional unit that can be manipulated.
Complexity of cell function is only part of why Cytomics will become a major field of study. Every cell is different. By studying each cell's unique function, that cell type can be further modeled for subsequent analysis using statistical techniques.
As the field of tissue engineering explodes, it will not be long before cellular engineering will be a most important component of which an essential element will be a full understanding of Cytomics.
Cytomic-realignment….
Within a short time, no pharmaceutical company will operate without encompassing the essential features of Cytomics
Drugs design will operate at the level of modified cellular functions, cytome-alignment or cytomic-realignment will become the "cellular form" of tissue engineering.
..how does the cell operate… This knowledge will require a better-than-ever
understanding of how the cell operates, how to measure cell function, and how to characterize the live cell in minute detail.
Single-cell analysis techniques will become enhanced and exquisitely sensitive.
New technologies must be developed and new analytical tools will be required to extract these new data.
Of these analytical tools, informatics will continue to play a crucial role in cell biology.
The big link…
Cytomics links technology to functional biology
Cytomics relates measurement & detection to structure & function
Cytomics integrates tools like flow cytometry, image cytometry, etc. with proteomics.
Cytomics….summary so far
Integration of technologies Functional role of system components Relates measurement & detection to structure &
function Brings together traditional cytometry and non
traditional cytometry Informatics now assumes a primary rather than a
secondary role in cytomics
Current Emphasis
gene
protein
cell
cell
protein
gene
protein
cell
gene
Live Cell
Hey buddy…Don’t you know you
genes, proteins and
organelles are in my territory
now!!
Slide animationis completePlease wait till slide animation is complete
Systems Integration
Analytical Cytology Flow cytometry Single cell analysis systems Tissue analysis (after cell separation)
Image Cytometry & Analysis Single cells Tissues and sections Cell culture systems 3D and 4D cell culture environments
Proteomics Proteins from specific cell populations Rapid identification
Imaging Technologies?
• DNA arrays
• “Quantitative” fluorescence assays
• High Throughput assays (96-384-1536 well plates)
• Elispot
• Drug effect assays, Cyto-toxicity
• Toxicology assays
• Apoptosis
• Cell proliferation, Cell ploidy
• Cytoplasm-to-nucleus transportation
• Hormone receptors, Growth factors
• Gene amplification or deletion, Gene fusion
• Chromosome imbalances
• And the list goes on……..
Next Generation Instruments…
40 fluorescent colors (40-50 variables & 100-200 parameters) Lots of other spatial measurements Lifetime Hyperspectral Imaging - Spectral unmixing/deconvolution technologies Multiple probe systems Complex analytical tools – informatics approaches 120,000 events/s for flow systems (4 x 108/hour) Very high speed for imaging systems Permanent and accurate alignment Intelligent interfaces and operating systems Direct links with diagnostic expert systems
This is HIGH Content
• Huge number of Variable & Parameters
• Very High Speed
• Huge data sets• Opportunity for Rapid classification
RapidIdentification
orDiagnostics
Much of this can becomeReal-Time decision making
Laser Scanning Cytometer
First “modern” high content static cell analysis system
Very high content Moderate speed Very high data
storage required Data-base friendly
Concept first published: Kamentsky & Kamentsky, Cytometry 12:381-7, 1991
Many Spectra in Flow Cytometry
Roederer, et al
Multi-Component Systems Amnis Corp
http://www.amnis.com
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Future integration
Eprogen-Beckman-Coulter automated protein separation system
Cytomics Jia Lu Kathy Ragheb Cheryl Holdman Gretchen Lawler
Additional Material for Discussion
The following materials were incorporated to highlight a number of educational facilities.
Appendix
Some Key Web Linkswww.cyto.purdue.eduhttp://www.cyto.purdue.edu/HCS
Issues that can be addressed:
1. Discussion Group - Communication2. Educational – knowledge development and training3. Issues of data file standards4. Issues of calibration standards5. Image processing issues – algorithms and processes
Opportunity?Discovery Park, Purdue UniversityCenter for Applied Cytomics
Expanded educational roleCan we partner with HCS users and developersCan we train in the basic issues
ISAC International Society for Analytical
Cytology Forum to address issues Cell-focused – understands “high
content” Has an existing structure International meeting May 23-28, 2004 in