Open AccessFLOW CYTOMETRIC CELL CYCLE ANALYSIS USING THE QUENCHING OF 33258 HOECHST FLUORESCENCE BY BROMODEOXYURIDINE INCORPORATION
Author(s) -
Bohmer R. M.
Publication year - 1979
Publication title -
cell proliferation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.647
H-Index - 74
eISSN - 1365-2184
pISSN - 0960-7722
DOI - 10.1111/j.1365-2184.1979.tb00117.x
Subject(s) - bromodeoxyuridine , thymidine , fluorescence , quenching (fluorescence) , dna , cell cycle , flow cytometry , microbiology and biotechnology , dna synthesis , biology , cell division , chemistry , cell growth , biophysics , cell , biochemistry , physics , quantum mechanics
Cultures of L cells were grown in medium containing 2.0 mg/l bromodeoxyuridine (BUdR) and stained with the fluorescent dye 33258 Hoechst for flow cytometric analysis. During exposure to BUdR, the cells replace thymidine by BUdR in the newly synthesized DNA. The new DNA is not stainable with 33258 Hoechst, which is highly specific for thymidine. The temporal development of the fluorescence distributions after addition of BUdR to the growth medium has been investigated in the flow cytometer, and the data were used to calculate the mean durations of the phases G 1 , S and G 2 + M in exponentially growing cultures as well as the cycle transit times in synchronized cultures. The percentage of non‐cycling cells was determined in each experiment.