Mismating is one of the most frequently presented reproductive problems presented to veterinarians. Confirming whether or not a mismating has occurred is necessary to determine if therapy will be instituted. Detection of spermatozoa in the vagina is irrefutable evidence of copulation; however, absence of sperm in a vaginal cytology cannot rule out coitus. In order to improve detection of spermatozoa post-coitus, a prospective study was initiated utilizing natural breedings of purebred beagles.