title: Assessment of Telomere Length, Phenotype, and DNA Content.
abstract: Telomere sequences at the end of chromosomes control somatic cell division; therefore, telomere length in a given cell population provides information about its replication potential. This unit describes a method for flow cytometric measurement of telomere length in subpopulations using fluorescence in situ hybridization of fluorescently-labeled probes (Flow-FISH) without prior cell separation. After cells are stained for surface immunofluorescence, antigen-antibody complexes are covalently cross-linked onto cell membranes before FISH with a telomere-specific probe. Cells with long telomeres are included as internal standards. Addition of a DNA dye permits exclusion of proliferating cells during data analysis. DNA ploidy measurements of cells of interest and internal standard are performed on separate aliquots in parallel to Flow-FISH. Telomere fluorescence of G0/1cells of subpopulations and internal standards obtained from Flow-FISH are normalized for DNA ploidy, and telomere length in subsets of interest is expressed as a fraction of the internal standard telomere length. © 2017 by John Wiley & Sons, Inc.
authors.affiliation: Department of Medicine, Division of Infectious Diseases, David Geffen School of Medicine at UCLA, Los Angeles, California.
authors.forename: Theodoros
authors.initials: T
authors.lastname: Kelesidis
authors.affiliation: Department of Medicine, Hematology/Oncology, David Geffen School of Medicine at UCLA, Los Angeles, California.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Schmid
mesh_terms: D000818:Animals; D004247:DNA / Q000032:analysis*; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique; D006801:Humans; D016130:Immunophenotyping; D017404:In Situ Hybridization, Fluorescence; D010641:Phenotype; D011003:Ploidies; D016615:Telomere / Q000737:chemistry*
chemical_list: D004247:DNA
keywords: Flow-FISH; cell surface immunofluorescence; fluorescence in situ hybridization; telomere length; telomeres