Flow cytometry is a sensitive and quantitative platform for the measurement of particle fluorescence. In flow cytometry, the particles in a sample flow in single file through a focused laser beam at rates of hundreds to thousands of particles per second. During the time each particle is in the laser beam, on the order of ten microseconds, one or more fluorescent dyes associated with that particle are excited. The fluorescence emitted from each particle is collected through a microscope objective, spectrally filtered, and detected with photomultiplier tubes. Flow cytometry is uniquely capable of the precise and quantitative molecular analysis of genomic sequence information, interactions between purified biomolecules and cellular function. Combined with automated sample handling for increased sample throughput, these features make flow cytometry a versatile platform with applications at many stages of drug discovery. Traditionally, the particles studied are cells, especially blood cells; flow cytometry is used extensively in immunology. This volume shows how flow cytometry is integrated into modern biotechnology, dealing with issues of throughput, content, sensitivity, and high throughput informatics with applications in genomics, proteomics and protein-protein interactions, drug discovery, vaccine development, plant and reproductive biology, pharmacology and toxicology, cell-cell interactions and protein engineering.
Author(s): Larry A. Sklar
Publisher: Oxford University Press, USA
Year: 2005
Language: English
Pages: 401
Contents......Page 6
Contributors......Page 10
1. The Future of Flow Cytometry in Biotechnology......Page 16
Part I: Tools for Flow Cytometry......Page 26
2. Fluorescent Probes......Page 28
3. Automation and High-Throughput Flow Cytometry......Page 53
4. Multispectral Imaging in Flow: A Technique for Advanced Cellular Studies......Page 70
5. Elastomeric Microfabricated Fluorescence-Activated Cell Sorters......Page 81
6. Flow Cytometry, Beads, and Microchannels......Page 118
7. DNA Fragment Sizing by High-Sensitivity Flow Cytometry: Applications in Bacterial Identification......Page 136
8. A Guide to Informatics and Flow Cytometry......Page 152
9. Molecular Assemblies, Probes, and Proteomics in Flow Cytometry......Page 166
Part II: Biological Applications......Page 192
10. Multiparameter Analysis: Application to Vaccine Analysis......Page 194
11. FACS-Based High-Throughput Functional Screening of Genetic Libraries for Drug Target Discovery......Page 206
12. Applications of Flow Cytometry in Protein Engineering......Page 225
13. Applications of Flow Cytometry in Animal Reproduction......Page 243
14. Genetic Analysis Using Microsphere Arrays......Page 272
15. Uses of Flow Cytometry in Preclinical Safety Pharmacology and Toxicology......Page 288
16. Flow Cytometry and Cell Sorting in Plant Biotechnology......Page 304
17. Analysis of GTP-Binding Protein–Coupled Receptor Assemblies by Flow Cytometry......Page 336
18. Applications of Flow Cytometry to Cell Adhesion Biology: From Aggregates to Drug Discovery......Page 360
A......Page 388
C......Page 389
D......Page 390
E......Page 391
F......Page 392
G......Page 393
I......Page 394
M......Page 395
P......Page 397
S......Page 399
T......Page 400
Z......Page 401
