The use of the stain exclusion assay and the hypoosmotic swelling test in determining the viability of human sperm was compared in sperm samples obtained from a group of 11 healthy males. Untreated sperm was compared with sperm treated with 2 percent glutaraldehyde (GA) or 0.1 percent Triton-X-100 (TX). Almost all sperm were non viable in the TX treated samples by both the stain exclusion assay and the hypoosmotic swelling test, as compared to only about 50 percent of the sperm in the control samples. Most of the sperm were non viable in the hypoosmotic test following treatment with GA, while the stain exclusion test failed to reveal any differences between GA treated and control sperm. Scanning and transmission electron microscopy tests indicated swelling of the sperm membrane tail and repeated coiling of the tail fibers within the swollen membrane in the hypoosmotic swelling test. The authors conclude that since the stain exclusion test is based on measurement of the structural integrity of the sperm membrane only, while the hypoosmotic swelling test is indicative of its physiological integrity, the two tests examined evaluate different characteristics of the sperm membrane.
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