Open AccessRAPID CELL CYCLE ANALYSIS BY MEASUREMENT OF THE RADIOACTIVITY PER CELL IN A NARROW WINDOW IN S PHASE (RCS i )
Author(s) -
Gray J. W.,
Carver June H.,
George Yolanda S.,
Mendelsohn M. L.
Publication year - 1977
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.1365-2184.1977.tb00135.x
Subject(s) - cell cycle , flow cytometry , chinese hamster ovary cell , mitosis , cell , phase (matter) , cell counting , biology , dna , cell division , microbiology and biotechnology , chemistry , biophysics , cell culture , biochemistry , genetics , organic chemistry
A new rapid method for the cell cycle analysis of asynchronously growing cells is presented. The new method is an alternative to the more time consuming and subjective fraction of labeled mitoses (FLM) method. Like the FLM method, all cells in the S phase of the cell cycle are marked by pulse labeling with a radioactive DNA precursor. The subsequent progress of the cohort of cells thus labeled is monitored through a narrow window in the cell cycle. The window is defined by a narrow range of DNA contents corresponding to cells in mid‐S phase and is designated S i . The cellular DNA content is measured by flow cytometry and the cells in the window S i are selected by electronic cell sorting. The radioactivity per cell in S i (RCS i ) is determined by liquid scintillation counting. The duration of S phase and of the total cycle and the dispersions therein are determined from the oscillation of the RCS i values with time. The complete cell cycle analysis can be accomplished in as little as 1 day following the collection of samples. Exponentially growing Chinese hamster ovary (CHO) cells were analyzed according to the RCS i method and the FLM method. It is demonstrated that the two techniques give essentially the same results.