The application of eosin and propidium iodide in evaluation of vitality of human spermatozoa
The article analyzes comparative assessment of vitality of spermatozoa by condition ofpermeability of membranes for eosin and propidium iodide and comparison of results acquired using technique of light and fluorescent microscopy. The comparison of data of light microscopy with eosin staining with data of fluorescent microscopy with propidium iodide staining demonstrated that percentage of content of spermatozoa separated from ejaculates of 28 fertile males and stained with eosin was reliably higher (34.8±3.2) than percentage of content of spermatozoa with stained with propidium iodide (21.1±4.0). After incubation of spermatozoa under room temperature during 24 hours percentage of unviable cells with stained eosin also was higher than in case of propidium iodide staining correspondingly (44.5±3.3% and 34.7±3.6%). The analysis of vitality of spermatozoa under damaging effect of oxidative stress on cell membrane developed by 4 hours incubation with 200 mkM of hydrogen peroxide (H2O2) demonstrated that under staining of spermatozoa with propidium iodide significantly higher percentage of damaged cells is detected. In such cases, eosin staining is less suitable for detection of vitality of spermatozoa (73.6±5.8% against 51.7±6.4%). The carried out experiment demonstrates that in case of detected effects on spermatozoa (for example, effect of oxidative stress) the light microscopy insufficiently adequate reflects degree of damage of membranes of spermatozoa. The fluorescent microscopy detects a higher percentage of spermatozoa with damaged membrane.