Essential Cytometry Methods

Cytometry is characterization and measurement of cells and cellular constituents, most often used to immunophenotype cells - that is, to distinguish healthy cells from diseased cells. Flow Cytometry specifically is quite sensitive, allowing researchers to detect rare cell types and residual levels of disease, and as such has been the method of choice for important studies such as monitoring the blood of AIDS patients. For this reason, there is a great need for a practical, comprehensive manual that will be useful across a broad range of laboratories. This volume, as part of the Reliable Lab Solution Series, delivers such a tool, offering busy researchers across many disciplines a handy resource of all the best methods and protocols for Cytometry to use at the bench.

* Highlights top downloaded and cited chapters, authored by pioneers in the field and enhanced with their tips, and pitfalls to avoid.  * Loaded with detailed protocols developed and used by leaders in the field. *Refines, organizes and updates popular methods from one of our top selling series, Methods in Cell Biology

Part I: Fluorochromes/General Techniques 1. Principles of confocal microscopy (Robinson) 2. Protein labeling with fluorescent probes (Holmes, Lantz) 3. Cytometry of fluorescence resonance energy transfer (Vereb, Matko, Szollosi) 4. The rainbow of fluorescent proteins (Galbraith) 5. Labeling cellular targets with semiconductor quantum dot conjugates (Wu, Bruchez) 6. High-gradient magnetic cell sorting (Radbruch, Mechtold, Thiel, Miltenyi, Pfluger) 7. Multiplexed microsphere assays for protein and DNA binding reactions (Kellar, Oliver) 8. Biohazard sorting (Schmid, Roederer, Koup, Ambrozak, Perfetto) 9. Guidelines for the presentation of flow cytometric data. (Roederer, Darzynkiewicz, Paks) Part II: Cellular DNA Content Analysis 10. VinDetergent and proteolytic enzyme-based techniques for nuclear isolation and DNA content analysis.(Vindelov,Christensen) 11. Rapid DNA content analysis (Krishan) 12. DNA analysis from paraffin-embedded blocks. (Hedley) 13. Flow cytometry and sorting of plant protoplasts and cells (Galbraith) 14. DNA content histogram and cell cycle analysis. (Rabinovitch) 15. Simultaneous analysis of cellular RNA and DNA content (Darzynkiewicz) Part III. Cell Proliferation and Death Assays 16. Immunochemical quantitation of bromodeoxyuridine: application to cell cycle kinetics (Dolbeare, Selden)| 17. Cell cycle kinetics estimated by analysis of bromodeoxyuridine incorporation (Terry, White) 18. Flow cytometric analysis of cell division history using dilution of carboxyfluorescein diacetate succinimidyl ester, a stably integrated fluorescent probe. (Lyons, Hasbold, Hodgkin) 19. Antibodies against the Ki-67 protein: Assessment of the growth fraction and tools for cell cycle analysis (Endl, Hollman, Gerdes) 20. Detection of DNA damage in individual cells by analysis of histone H2AX phosphorylation (Olive) 21. Assays of cell viability: Discrimination of cells dying by apoptosis. (Darzynkiewicz, Li, Gong) 22. Difficulties and pitfalls in analysis of apoptosis (Darzynkiewicz, Bedner, Traganos) Part IV: Cell Surface Immunophenotyping 23. Cell preparation for the identification of leukocytes (Stewart, Stewart) 24. Multicolor immunophenotyping: Human immune system hematopoiesis (Wood) 25. Differential diagnosis of T-cell lymphoproliferative disorders by flow cytometry multicolor immunophenotyping. Correlation with morphology. (Gorczyca) 26. B-cell immunophenotyping (Baumgarth) Part V: Cytogenetics/Chromatin Structure 27. Telomere length measurements using in situ hybridization(Baerlocher, Lansdorp) 28. Sperm chromatin structure assay: DNA denaturability (Evanson, Jost) Part VI: Cell Physiology Assays 29. Cell membrane potential analysis. (Shapiro) 30. Measurement of intracellular pH. (Boyer, Hedley) 31. Intracellular ionized calcium (June, Rabinovitch) 32. Oxidative product formation analysis by flow cytometry (Robinson, Carter, Narayanan). 33. Phagocyte function. (Rothe, Glouche) 34. Analysis of RNA synthesis by cytometry (Jensen, Larsen, Larsen) 35. Analysis of mitochondria by flow cytometry (Poot, Pierce) 36. Analysis of platelets by flow cytometry (Ault, Mitchell) Part VII: Detection of Microorganisms and Pathogens 37. Detection of specific microorganisms in environmental samples using flow cytometry (Vesey, Narai, Ashbolt, Williams, Veal) 38. Flow cytometric analysis of microorganisms. (Sincock, Robinson) 39. Flow cytometry of malaria detection (Janse, Van Vianen)