Methods commonly used for detection of DNA double-strand breaks (DSBs) and analysis of cell death are generally time-consuming, and, therefore, any improvements in these techniques are important for researchers and clinicians. At present, flow cytometry is the most rapid method for detection of DSBs and cell viability. In this chapter, we provide our experience and methodological modification of flow cytometry protocol for the detection of γ-H2AX, a well-known marker of DSBs, in fixed mammalian fibroblasts. The modifications permit a reduction in the time required for DSB detection by flow cytometry.
Keywords: DNA double-strand breaks; Fibroblasts; Flow cytometry; Rapid detection; γ-H2AX.