Chapter 16 Cell cycle kinetics estimated by analysis of Bromodeoxyuridine Incorporation

Publisher Summary This chapter describes the methods for sample preparation and staining, flow-cytometric data acquisition, and the procedures for quantitative analysis of dynamic cell-kinetic data. Whereas the percentage of labeled mitotic figures (PLM) technology relied on labeling cells in S-phase and observing them through the window of mitosis, the two-color flow cytometric technique relies on S-phase labeling but now observes the labeled cells throughout the entire cell cycle. Importantly, it utilizes the extra information discernible from the division status of the labeled population. This technology permits the rapid evaluation of the quantities required to describe many of the growth kinetic parameters of cell populations. The basis of these techniques is that cells in S phase can be selectively labeled both in vitro and in vivo by administration of a nontoxic level of bromodeoxyuridine (BrdUrd). The cells that incorporate BrdUrd continue to progress through the cell cycle and may be sampled, or a biopsy or surgical specimen taken, at a known time later.