title: Measurement of intracellular ions by flow cytometry.
abstract: Using flow cytometry, single-cell measurements of calcium can be made on isolated populations identified by one or more phenotypic characteristics. Most earlier techniques for measuring cellular activation parameters determined the mean value for a population of cells, which did not permit optimal resolution of the responses. The flow cytometer is particularly useful for this purpose because it can measure ion concentrations in large numbers of single cells and thereby allows ion concentration to be correlated with other parameters such as immunophenotype and cell cycle stage. A limitation of flow cytometry, however, is that it does not permit resolution of certain complex kinetic responses such as cellular oscillatory responses. This unit describes the preparation of cells, including labeling with antibodies and with calcium probes, and discusses the principles of data analysis and interpretation.
authors.affiliation: Abramson Family Cancer Research Institute, and the Department of Pathology and Laboratory Medicine, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, Pennsylvania.
authors.forename: Avery D
authors.initials: AD
authors.lastname: Posey
authors.affiliation: Abramson Family Cancer Research Institute, and the Department of Pathology and Laboratory Medicine, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, Pennsylvania.
authors.forename: Omkar U
authors.initials: OU
authors.lastname: Kawalekar
authors.affiliation: Abramson Family Cancer Research Institute, and the Department of Pathology and Laboratory Medicine, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, Pennsylvania.
authors.forename: Carl H
authors.initials: CH
authors.lastname: June
mesh_terms: D000818:Animals; D002021:Buffers; D002118:Calcium / Q000378:metabolism; D002138:Calibration; D004533:Egtazic Acid / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D005453:Fluorescence; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism*; D007477:Ions; D051379:Mice; D020442:Poloxamer / Q000494:pharmacology; D013050:Spectrometry, Fluorescence
chemical_list: D002021:Buffers; D007477:Ions; D020442:Poloxamer; D004533:Egtazic Acid; D002118:Calcium
keywords: calcium analysis; flow cytometry; ion analysis