title: Application of flow-FISH for dynamic measurement of telomere length in cell division.
abstract: This method makes it possible to measure the fluorescence of a DNA probe in cells with known division number and targeted surface antigen. In fact, this method is a combination or consistent application of three other methods: cell tracking by vital dye, surface immunophenotyping, and flow-FISH. The idea in developing this method was to study telomere length changes in cells with known surface antigen after every new cell division. First, the in vitro cell culturing and staining with CFSE vital dye are performed. Then, cells are stained with surface MAbs labeled with biotin, followed by incubation with streptavidin-labeled fluorochrome. After that, cells are fixed with BS(3) reagent followed by the flow-FISH procedure with PNA-probe complementary to telomere DNA repeats. Finally, in one tube, it is possible to determine telomere length in surface antigen-labeled cells that have made the exact same number of divisions after incubation.
authors.affiliation: Research Institute of Clinical Immunology, Russian Academy of Medical Sciences, Novosibirsk, Russia.
authors.forename: Vyacheslav I
authors.initials: VI
authors.lastname: Borisov
authors.forename: Olga Y
authors.initials: OY
authors.lastname: Korolkova
authors.forename: Vladimir S
authors.initials: VS
authors.lastname: Kozhevnikov
mesh_terms: D000818:Animals; D002455:Cell Division / Q000502:physiology*; D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D016615:Telomere / Q000235:genetics / Q000378:metabolism*
chemical_list: D015342:DNA Probes; D005456:Fluorescent Dyes
keywords: CFSE; cell tracker; flow-FISH; telomere length; telomeres