Flow Cytometry

In flow cytometry, measurements of cells or other biological particles are made as the cells or particles flow in single file in a fluid stream past optical and/or electronic sensors. Modern flow cytometers typically use lasers as light sources and measure light scattered by cells, which provides information about their size and internal structure, and fluorescence in several spectral regions emitted by dyes or labeled probes or reagents which bind specifically and stoichiometrically to cellular constituents such as antigens and nucleic acids. Flow sorting allows cells with preselected characteristics to be diverted from the stream and collected for further analysis. The optics of a flow cytometer are similar to those of a fluorescence microscope.

Multiparameter flow cytometry, that is, the measurement of several characteristics of each cell or particle analyzed, permits precise characterization of multiple cell subpopulations in complex mixtures; signals from ten or more fluorescent labels have been used in combination with scattering measurements to define functional subsets of lymphocytes in instruments which analyze as many as 100,000 cells/second and can quantify fluorescence down to the level of a few hundred molecules. Using suitable fluorescent probes, it is possible to measure functional properties of cells, such as levels of gene expression and cytoplasmic and mitochondrial membrane potential. Some laboratory-built flow cytometers utilize slower analysis rates to increase detection sensitivity and precision, allowing measurements to be made of nucleic acid fragments and viruses.

References

Papers and Textbooks:

• Darzynkiewicz Z, Crissman HA, Robinson JP (Eds.): Flow Cytometry. 3rd Edition. Methods in Cell Biology, Volumes 63 (Part A) and 64 (Part B). San Diego, Academic Press, 2000.
• Diamond RA, DeMaggio S (Eds.): In Living Color: Protocols in Flow Cytometry and Cell Sorting, Berlin, Springer, 2000.
• Givan AL: Flow Cytometry: First Principles. 2nd Edition. New York, Wiley-Liss, 2001.
• Ormerod MG: Flow Cytometry. 2nd edition. Oxford, Bios, 1999.
• Ormerod MG (Ed.): Flow Cytometry - A Practical Approach. 3rd edition. Oxford, Oxford University Press, 2000.
• Robinson JP, Darzynkiewicz Z, Dean P, Dressler L, Rabinovitch P, Stewart C, Tanke H, Wheeless L, (Eds.): Current Protocols in Cytometry, New York, John Wiley & Sons (continuing updates).
• Shapiro HM: Practical Flow Cytometry. 3rd Edition. New York, Wiley-Liss, 1995.

Special Journal Volumes:

• Alberghina L, Porro D, Shapiro H, Srienc F, Steen H (Eds.): Microbial Analysis at the Single-Cell Level. Journal of Microbiological Methods, Volume 42, Number 1, 2000
• Lenkei R, Mandy F, Marti G, Vogt R (Eds.): Quantitative Fluorescence Cytometry: An Emerging Consensus. Cytometry, Volume 33, Number 2, October 1, 1998.
• Weaver J (Ed.): Flow Cytometry. Methods, Volume 21, 2000.
• Zola H (Ed.): Flow Cytometry. Journal of Immunological Methods, Volume 243, Numbers 1 and 2, 2000.