High Content Screening Science, Techniques and Applications

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ISBN: 9780470039991 | 047003999X
Cover: Hardcover
Copyright: 1/18/2008

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The authoritative reference on High Content Screening (HCS) in biological and pharmaceutical research, this guide covers: the basics of HCS: examples of HCS used in biological applications and early drug discovery, emphasizing oncology and neuroscience; the use of HCS across the drug development pipeline; and data management, data analysis, and systems biology, with guidelines for using large datasets. With an accompanying CD-ROM, this is the premier reference on HCS for researchers, lab managers, and graduate students.

Preface
Contributors
Essentials Of High Content Screening.
Approaching High Content Screening and Analysis: Practical Advice for Users
Introduction
What is HCS and Why Should I Care?
How does HCS Compare with Current Assay Methods?
The Basic Requirements to implement HCS
The Process
An Example Approach
Six Considerations for HCS Assays
References
Automated High Content Screening Microscopy
Introduction
Automated HCS Imaging Requirements
Components of Automated Imaging Platforms
Imaging Platform Software
Data Storage and Management
Selecting an HCS Platform
Comparison of a SAPK Activation HCS Assay Read on an ArrayScanw 3.1, an ArrayScanw VTi, and an IN Cell 3000 Automated Imaging Platform
References
A Primer on Image Informatics of High Content Screening
Background
HCS Image Processing
Validation
Information System Management
Data Modeling
Conclusions
Acknowledgements
References
Developing Robust High Content Assays
Introduction
Overview of a Typical Immunofluorescence-Based High Content Assay
Identifying Sources of Variability in a High Content Assay
From Immunofluorescence to High Content: Selecting the Right Metric
Validation of High Content Assays
Conclusion
Acknowledgements
References
Section II
HCS in Cellular Oncology and Tumor Biology
Cancer Cell Biology and HCS
The Cell Biology of Cell Death
Cell Signaling Pathways in Cancer
HCS in Tumor Biology
Conclusions
References
Exploring the Full Power of Combining High Throughput RNAi with High Content Readouts: From Target Discovery Screens to Drug Modifier Studies
Background: The Convergence of High Content Analysis and RNAi
Integrating HT-RNAi and HCA in Drug Discovery: The Potential
Combining RNAi and HCA in one Assay - the Reality
HCA-Based RNAi Studies - The Future
Acknowledgements
References
Leveraging HCS in Neuroscience Drug Discovery
High Content Screening and Drug Discovery
The Neuron and Neuronal Morphology
Methods for Measuring Neuronal Morphology
Small Molecule Screening for Neurite Outgrowth
RNAi in Neuroscience and HCA
Measurement of Signal Transduction in Neurons
High Content Screening in Complex CNS Models
Methods used in Neuronal HCS
References
Live Brain Slice Imaging for Ultra High Content Screening: Automated Fluorescent Microscopy to Study Neurodegenerative Diseases
Introduction and Background
Live Brain Slice Model to Study Huntington's Disease
Imaging Platforms
Center of Well (COW) for Image Processing
Generic Protocol for the Cellomics ArrayScan VTI
Data and Results
Discussion
References
High Content Analysis of Human Embryonic Stem Cell Growth and Differentiation
Introduction
Cell Culture Methods
Statistical Wavelet Analysis for Stem Cell Classification
Molecular Analysis of Pluripotency and Cell Proliferation in Undifferentiated Stem Cells
Analysis of Cardiomyocyte Differentiation
Analysis of Neuronal Differentiation
References
HCS IN DRUG DEVELOPMENT.
HCS for HTS
Introduction
HCS for Orphan GPCRS and Transfluor
HCS for Multiparameter Cytotoxicity Screening
Discussion
Summary
References
The Roles of High Content Cellular Imaging in Lead Optimization
Introduction
Statistical Validation of Assays
High Content Cellular Imaging is a Diverse Assay Platform
Use of High Content Cellular Imaging For Oncology Research at Eli Lilly
The Future of High Content Cellular Imaging in Lead Optimization
Acknowledgements
References
Using High Content Analysis for Pharmacodynamic Assays in Tissue
Introduction
Designing a High Content Assay for Use in Tissues
Technical Challenges in Establishing High Content Assays for Tissue
Case Study: Design and Validation of a High Content Assay for Biomarker X
Conclusions
Acknowledgements
References
High Content Analysis of Sublethal Cytotoxicity in Human HepG2 Hepatocytes for Assessing Potential and Mechanism for Chemicaland Drug-Induced Human Toxicity
Introduction
Results from High Content Analysis of Human Toxicity Potential
Discussion
Acknowledgements
Appendix: Detailed Methods
References
Data Management, Data Analysis And Systems Biology.
Open File Formats for High Content Analysis
Introduction
The Data Problem in Biology: Why is it so Hard?
High Content Data in Biology: A Definition
The difference Between a File Format and a Minimum Specification
File Formats: Open vs Closed
File Formats: Balancing Flexibility with Standards
Supporting a Successful File Format
Commercial Realities: How Users and Developers can Define File Formats
OME-ML and OME-TIFF: Moving Towards a Standard Format For High Content Biological Data
Data Model and File Format Integration: Towards Usable Tools
Conclusions
Acknowledgements
References
Analysis of Multiparametric HCS Data
Cytological Classification and Profiling
Setting Up Cytological Profiling Studies
Sources of Variability and Corrections
General Analysis Considerations
Data Analysis Methods
Software for HCS Data Analysis
Conclusions
References
Quantitative and Qualitative Cellular Genomics: High Content Analysis as an Endpoint for HT-RNAi Phenotype Profiling Using GE's InCell Platform
Cellular Genomics
Enabling Technologies to Facilitate Cellular Genomics: RNA Interference
High Throughput Rnai (HT-RNAi
High Content Analysis (HCA) for High Throughput Phenotype Profiling
Future Directions
References
Optimal Characteristics of Protein-Protein Interaction Biosensors for Cellular Systems Biology Profiling
Introduction
Challenge of Cellular Systems Biology (CSB
Optimal Characteristics of Protein-Protein Interaction Biosensors (PPIBs
Example of a PPIB and Cellular Systems Biology Profiling
Summary and Prospects
Acknowledgements
References
Index
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High Content Screening Science, Techniques and Applications

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