Sperm detection can be an important factor in confirming sexual assault in cases of rape. This paper compares three of the most commonly used staining methods cited in the scientific literature: Christmas tree, hematoxylin-eosin, and alkaline fuchsin. The population studied was composed of 174 consenting women seen at the Male Infertility Center in Toulouse, France. The date of their last sexual intercourse was accurately known. Alkaline fuchsin did not seem effective in detecting spermatozoa in vaginal samples. Compared with hematoxylin-eosin, Christmas tree stain appeared to be the most useful test in the first 72 h. Two external factors were associated with decreased detection of spermatozoa: time since intercourse and sperm volume.
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