Evaluating sperm cell viability and membrane integrity

Abstract

Rapid and precise assessment of sperm viability is vital in evaluating seminal quality. The viability (%) is determined by identifying sperm with an intact cell membrane, either by carrying out dye exclusion tests or by hypo-osmotic swelling. In this chapter, we have attempted to cover some important assays with the underlying principles including assay based on the hypotonic swelling and its variations as an indicator of the biochemical integrity of the membrane. This chapter contains procedures related to hypo osmotic swelling and application of supravital dye and various fluorescent dyes such as Hoechst 33342, DAPI, PI and SYBR-14 including their combination for determining spermatozoa viability. A comparison of the assay vis-à-vis their merit has been provided.