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Abstract
The multiple-layer discontinuous Percoll density gradient centriftigation procedure is being used for gender selection and several reports suggested separation efficiencies of over 77%. The mechanism involved in the separation of X- and Y-bearing sperm using this method seems to be the difference in sperm head dimensions or motility but supporting data are inconsistent. The specific aims of the study were to evaluate the head dimensions of sperm at the upper and lower fractions after the 8-layer Percoll gradient procedure for sex preselection and to ascertain the kinematic parameters and tail lengths of sperm derived from the 2 separate Percoll fractions. Sperm cells were obtained from thawed donor specimens (A' = 20) and were layered on top of the 8-layer discontinuous Percoll gradient, which ranged from 34 to 85% in increments of 7%. After centrifugation, the resuspended sperm cells derived from the upper and lower fractions of the Percoll gradient were analyzed on the Hamilton Thorn HTM-C analyzer for differences in sperm motility patterns and sperm head dimensions. Aliquots of sperm from the 2 fractions were fixed and stained using the Spermac stain, and the lengths of each sperm tail (N = 600) were measured on the HTM-C analyzer.