title: Assaying Cell Cycle Status Using Flow Cytometry.
abstract: In this unit, two protocols are described for analyzing cell cycle status using flow cytometry. The first is based on the simultaneous analysis of proliferation-specific marker (Ki-67) and cellular DNA content, which discriminate resting/quiescent cell populations (G0 cell) and quantify cell cycle distribution (G1, S, or G2/M), respectively. The second is based on differential staining of DNA and RNA through co-staining of Hoechst 33342 and Pyronin Y, which is also useful to identify G0 cells from G1 cells. Along with these methods for analyzing cell cycle status, two additional methods for cell proliferation assays with recent updates of newly developed fluorophores, which allow multiplex analysis of cell cycle status, cell proliferation, and a gene of interest using flow cytometry, are outlined.
authors.affiliation: Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas.
authors.forename: Kang Ho
authors.initials: KH
authors.lastname: Kim
authors.affiliation: Cytometry and Cell Sorting Core, Baylor College of Medicine, Houston, Texas.
authors.forename: Joel M
authors.initials: JM
authors.lastname: Sederstrom
mesh_terms: D000818:Animals; D002453:Cell Cycle; D003584:Cytological Techniques / Q000379:methods*; D004247:DNA / Q000032:analysis; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D019394:Ki-67 Antigen / Q000032:analysis; D012313:RNA / Q000032:analysis; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D019394:Ki-67 Antigen; D012313:RNA; D004247:DNA
keywords: Hoechst 33342; Ki-67; Pyronin Y; cell cycle; flow cytometry; propidium iodide