Sperm aneuploidy testing using fluorescence in situ hybridization.

Sperm aneuploidy screening has been used as a tool in diagnosis and determining treatment options for male factor infertility since the development of human sperm karyotyping by injection into hamster and mouse oocytes in the 1970s. From these studies and subsequent work with interphase chromosome analysis, at risk populations of men with teratozoospermia, oligozoospermia, and men with translocations, have since been identified. The current technique is an application of fluorescent in situ hybridization (FISH) on interphase sperm nuclei with careful enumeration of the labeled chromosomes to determine sperm ploidy. Typically, five to seven chromosomes are evaluated in individual ejaculates to determine the percent of aneuploid sperm present. This protocol will detail the procedures for: preparation of specimens, exposure of the sperm nuclei to the FISH probes, hybridization, destaining, and scoring criteria.