Reliability of estimates of diploid human spermatozoa using multicolour fluorescence in-situ hybridization [published erratum appears in Hum Reprod 1997 May;12(5):1118]

Multicolour fluorescence in-situ hybridization (FISH) analysis permitsdistinction between disomic and diploid spermatozoa. Thus estimates of thefrequency of diploid spermatozoa can be obtained for human semen samples.The issue of the accuracy and reliability of these diploidy estimates hasbeen addressed by analysing diploidy frequencies in 10 men using the samesperm sample to estimate diploidy twice-once during two-colour FISHanalysis of disomy for chromosomes 1 and 12 and a second independentanalysis of three-colour FISH for disomy estimates for chromosomes X and Y(with chromosome 1 used as the autosomal control). A minimum of 10,000spermatozoa per hybridization per male was counted for a total of over200,000 spermatozoa analysed. The mean frequency of diploid spermatozoa was0.13% for the autosomal study and 0.14% for the sex chromosomal study,which were not significantly different. One donor had extremely divergentvalues of diploidy in the two studies. Analysis of values in the other ninedonors demonstrated no significant difference in the two diploidyestimates. These results indicate that the FISH technique is an accurateand reliable method for determining diploid frequencies in humanspermatozoa.