A method to measure the duration of DNA synthesis and the potential doubling time from a single sample.
A method is described whereby the DNA synthesis time, Ts, can be calculated using data of a single sample of cells taken several hours after labelling with bromodeoxyuridine (BrdUrd). The method involves a simple calculation using flow cytometry data of BrdUrd incorporation (green fluorescence, FITC-labelled anti-BrdUrd-DNA antibody) and total DNA content (red fluorescence, propidium iodide). The movement of BrdUrd-labelled cells through the S phase can be quantified by measuring their mean red fluorescence relative to that of G1 and G2 cells. Assuming the movement of the labelled cells toward G2 is linear with time, Ts can be calculated by measuring their relative movement at any one time. The method was tested on cells in vitro and on bone marrow and tumor cells in vivo. Reasonable agreement was seen with published estimates of Ts for these tissues.
D000818:Animals; D000911:Antibodies, Monoclonal; D001854:Bone Marrow Cells; D001973:Bromodeoxyuridine; D002453:Cell Cycle; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000032:analysis / Q000096:biosynthesis*; D005434:Flow Cytometry; D007399:Interphase; D051379:Mice
D000911:Antibodies, Monoclonal; D004247:DNA; D001973:Bromodeoxyuridine