Evaluation of Sperm Quality by Flow Cytometry: Sequential Protocols for DNA Fragmentation and Acrosome Reaction in Membrane-Intact Spermatozoa.

Evaluation of sperm DNA fragmentation and membrane integrity plays a critical role in understanding male fertility potential. Flow cytometry serves as a powerful and precise technique for this purpose. Our group has developed a convenient, reliable, and reproducible flow cytometry-based strategy for simultaneous evaluation of DNA and Membrane integrity (DMI). Building on this, we are currently conducting in vitro research on acrosome reaction analysis in live spermatozoa, an additional sperm membrane integrity-related parameter, using flow cytometry. This chapter presents both approaches as complementary protocols that can be run side by side: simultaneous evaluation of DNA and membrane integrity (DMI), and assessment of in vitro acrosome reaction and membrane integrity.