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      Analysis of cell cycle by flow cytometry.

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          Abstract

          Described are four widely used procedures to analyze the cell cycle by flow cytometry. The first two are based on univariate analysis of cellular DNA content following cell staining with either propidium iodide (PI) or 4',6'-diamidino-2-phenylindole (DAPI) and deconvolution of the cellular DNA content frequency histograms. This approach reveals distribution of cells in three major phases of the cycle (G1 vs S vs G2/M) and makes it possible to detect apoptotic cells with fractional DNA content. The third approach is based on the bivariate analysis of DNA content and proliferation-associated proteins. The expression of cyclin D, cyclin E, cyclin A, or cyclin B1 vs DNA content is presented as an example. This approach allows one to distinguish, for example, G0 from G1 cells, identify mitotic cells, or relate expression of other intracellular proteins to the cell cycle position. The fourth procedure relies on the detection of 5'-bromo-2'-deoxyuridine (BrdU) incorporation to label the DNA-replicating cells.

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          Author and article information

          Journal
          Methods Mol Biol
          Methods in molecular biology (Clifton, N.J.)
          Springer Science and Business Media LLC
          1064-3745
          1064-3745
          2004
          : 281
          Affiliations
          [1 ] Department of Clinical Immunology, School of Medicine, Lublin, Poland.
          Article
          1-59259-811-0:301
          10.1385/1-59259-811-0:301
          15220539
          13e27b3f-b0a8-4a22-a19f-d2a357f56bd4
          History

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