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

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. Copyright © 1989 Wiley‐Liss, Inc.