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Continuous bromodeoxyuridine labeling and bivariate ethidium bromide/hoechst flow cytometry in cell kinetics

1989; Wiley; Volume: 10; Issue: 2 Linguagem: Inglês

10.1002/cyto.990100215

1097-0320

Martin Poot, Heidi Schmitt, Helga Seyschab, Julia Koehler, Una Chen, U. Kaempf, Manfred Kubbies, Detlev Schindler, Peter S. Rabinovitch, Holger Hoehn,

Cell Image Analysis Techniques

Abstract Most techniques of flow cytometric cell cycle analysis are not capable of distinguishing the number of rounds of DNA synthesis that a cell has undergone since the start of an experiment. Continuous labeling with 5‐bromodeoxyuridine (Brd‐Urd) offers such a potential. We illustrate here that the bivariate analysis of non‐BrdUrd‐quenched ethidium bromide vs. BrdUrd‐quenched Hoechst 33258 fluorescence offers a high degree of resolution that enhances the analytical power of the technique, and that this approach can be applied to the analysis of a broad range of human and murine primary cells and established cell lines.