A comparison of time-lapse cinemicrography and flow cytometry for the study of accelerated cell-cycle transit

Abstract. Two methods for the study of cell-cycle progression, time-lapse cinemicrography (TLCM) and flow cytometry (FCM), were compared for their ability to measure the shortening of cell-cycle transit time induced by temporary inhibition of DNA synthesis. DNA synthesis was reversibly inhibited by aphidicolin (APH) in synchronized HeLa cells obtained by mitotic collection. TLCM directly measured intermitotic time intervals and thereby directly obtained the cell-cycle transit time distribution. In contrast, FCM measured time dependent changes in the fractions of cells in the cell-cycle phases from which the distribution of cells traversing a cell-cycle boundary, such as that between G₁ and S phase, was determined. Nevertheless, both methods provided equivalent measures of the cell-cycle transit time and its dispersion. However, TLCM apeared to provide a better measure of skewness of the transit time distribution than did FCM. Further, both methods were able to detect changes in the cell cycle transit on the order of 1 h or less. The TLCM data showed a greater precision (due to a larger number of data points) than that from FCM. However, FCM was able to directly measure changes in the transit of G₁ phase whereas TLCM would require two different experiments to make a similar determination. The results obtained in this study show that FCM can replace TLCM to study most aspects of cell-cycle progression.