A method to measure the duration of DNA syntheses and the potential doubling time from a single sample

AC Begg, NJ McNally, DC Shrieve… - Cytometry: The Journal …, 1985 - Wiley Online Library
AC Begg, NJ McNally, DC Shrieve, H Kärche
Cytometry: The Journal of the International Society for Analytical …, 1985Wiley Online Library
A method is described whereby the DNA synthesis time, Ts, call 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 Ouantified by measuring their mean red fluorescence relative to …
Abstract
A method is described whereby the DNA synthesis time, Ts, call 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 Ouantified 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, T., 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.
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