Flow cytometry in assessment of sperm integrity and functionality – a review

Abstract

Spermatozoa must possess many attributes to fertilize an egg but few laboratory methods can assess all of these attributes simultaneously and objectively. Most laboratory methods used to assess the quality of semen in veterinary andrology can be inaccurate and time-consuming. Laboratory techniques which evaluate only one sperm attribute, often provide results that have a weak correlation with fertility. Therefore, semen used for artificial insemination should not be assessed based on the results of one method only but rather on the comprehensive results of several laboratory tests. Flow cytometry is a modern method of analysing diJ169fferent types of cells, including sperm cells. It is based on the scattering of light and fluorescence, the outcome of which yields results that are recorded and evaluated by computer technology which makes an objective assessment. Flow cytometry in combination with fluorescence microscopy and fluorescent probes provides a comprehensive, accurate, objective, and rapid analysis of the ejaculate. In a short time frame it allows us to test thousands of sperm for their structure and properties, even with a minimal amount of semen. This method enables the evaluation of several indicators simultaneously in a population of sperm as a whole or for each sperm individually. It informs us about the selected indicators of sperm quality in the sample by examining the membrane integrity, DNA, mitochondria, acrosome, oxidative stress, and other properties. Flow cytometry has become an important method of evaluating the functional and morphological properties of sperm and is used for research in veterinary science as well as for a routine assessment of the semen quality.