Flow cytometry multiparameter analysis

Burchiel SW, Edwards BS, Kuckuck FW, Lauer FT, Prossnitz ER, et al. 2000. Analysis of free intracellular calcium by flow cytometry Multiparameter and pharmacologic applications. Methods 21 221-230. 

Scientific progress has resulted in the continual development and evolution of useful and powerful techniques, such as multiparameter flow cytometry and image analysis, to answer questions of cellular maintenance and function (and the impact chemicals may exert) for the benefit of human health. Multiparameter flow cytometry provides researchers with a capacity to simultaneously examine multiple cellular characteristics on thousands of cells in a relatively short period of time. 

The following basic protocols may be used alone or combined with other staining procedures in multiparameter flow cytometry experiments. Although they are illustrated with data from cells that proliferate in suspension, these protocols may be easily modified for the analysis of cells isolated from tissues or adherent cells in culture, by incorporating an initial step for the preparation of single cell suspensions. The assays are conducted at room temperature, unless otherwise noted. 

Dive, C., Gregory, C D., Phipps, D J, Evans, D L, Milner, A E, and Wylhe, A H (1992) Analysis and discrimination of necrosis and apoptosis by multiparameter flow cytometry Biochim Biophys Acta 1133, 275—285. 

The second aspect of clinical practice that has led to a reassessment of the nature of flow cytometry is the occasional clinical requirement for rare-event analysis. Methods have been developed, particularly with the use of multiparameter gating, to lower background noise in order to provide increased sensitivity for detection of rare cells. In the clinic, this increased sensitivity translates, for example, into earlier diagnosis of relapse in leukemia, more sensitive detection of fetal-maternal hemorrhage, and better ability to screen leukocyte-reduced blood transfusion products for residual white blood cells. Outside the clinic, these methods for rare-event detection have begun to stretch the limits of research applications as well. 

Petrausch U, Haley D, Miller W, Floyd K, Urba WJ, Walker E. Polychromatic flow cytometry a rapid method for the reduction and analysis of complex multiparameter data. Cytometry 2006 69A(12) 1162-1173. 

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