RAPID CELL CYCLE ANALYSIS BY MEASUREMENT OF THE RADIOACTIVITY PER CELL IN A NARROW WINDOW IN S PHASE (RCSi)

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Abstract
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 Si. The cellular DNA content is measured by flow cytometry and the cells in the window Si are selected by electronic cell sorting. The radioactivity per cell in Si (RCSi) 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 RCSi 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 RCSi method and the FLM method. It is demonstrated that the two techniques give essentially the same results.