Lab 3 - Analysis of the DNA Content of Cells by Single-Color Flow Cytometry

Alice Givan

 

Because the DNA content of cells is tightly regulated, flow analysis of DNA benefits from careful and informed use of the flow cytometer.  In this laboratory, we will discuss ways to stain cells for their DNA  content, protocols for using the cytometer effectively, and methods for analyzing flow data to provide information about the distribution of cells through the cell cycle.  We will emphasize factors that make DNA analysis ambiguous: particularly, high flow rates, aggregated cells, overlap of S-phase with other phases, and endoreduplication. We will leave students some appreciation for the ways in which flow cytometrists can cope with these ambiguities.