Flow Cytometric Determination of Cell Cycle Parameters of V79 Cells by Continuous Labeling with Bromodeoxyuridine

Abstract

A simple method with which to determine the cell cycle parameters, T_G1, T_S and T_G2M (the durations of the G₁, S and G₂+M phases) is described. V79 Chinese hamster lung cells were used to evaluate the method. After continuous labeling with bromodeoxyuridine (BrdU), V79 cells were stained with anti BrdU-monoclonal antibody with FITC (fluorescein isothio-cyanate) and with PI (propidium iodide). The individual cells were checked by flow cytometry for green and red fluorescences whose signal intensities cor-responded to the BrdU and cellular DNA contents. The durations of G₁, S and G₂+M phases of V79 cells were determined by measuring the cell fractions containing the nonlabeled G₁, labeled S and nonlabeled G₂+M phases. The reliability of this method is discussed.