The hypo-osmotic swelling test: Is it a sperm vitality or a viability assay?

The first two editions of the World Health Organization laboratory manual described the determination of live spermatozoa by a dye exclusion method as a sperm "viability" test, whereas subsequent editions classified it as a "vitality" test, without providing an explanation for the reclassification. Additionally, the hypo-osmotic swelling (HOS) test, which assesses the functional integrity of the human sperm membrane, was placed in the same category as the dye exclusion test. Although the two terms might seem synonymous, the term "vitality" merely means "alive," whereas "viability" assesses qualities or physiological functions of a living entity. After comparing the morphological, physiological, and clinical findings obtained from dye exclusion testing vs. the HOS test, we conclude that the HOS test should be classified as a viability test, not merely as a vitality test.