ABSTRACT
This chapter discusses the most widely used methods of DNA content analysis applied to cell-cycle analysis, as well as, the up-to-date cell-based fluorescent methodologies that are highly impacting on cell-cycle profiling. DNA analyses have always been performed by profiling the DNA content of individual cells from cell preparations or tissue specimens. DNA quantification allows information concerning the distribution of cells from a cellular population across the different cell-cycle phases, the quantification of apoptosis by the estimation of the fractional DNA, and it establishes the DNA ploidy status of cells. The chapter provides a snapshot of the distribution/frequency of cells in different cell-cycle phases, though, when obtained at different times, DNA histograms yield information on cell-cycle dynamics. It considers the cell type used and its applications, the DNA-specific fluorophore chosen, and the spectral characteristics of each fluorescent molecule in multiplexed analysis.
