Separation of human X- and Y-bearing sperm using percoll density gradient centrifugation

Human X- and Y-bearing sperm, which were detected by the quinacrine mustard fluorescent staining method, were separated in density gradient centrifugation using Percoll (Pharmacia Fine Chemicals, Uppsala, Sweden). The apparent percentages of Y-bearing sperm in normo- and oligospermia were 46.7% +/- 3.4% (n = 13) and 46.6% +/- 1.2% (n = 10), respectively. The separation was observed to some extent in Ficoll Paque (Pharmacia Fine Chemicals); then it was further achieved using Percoll. After centrifugation, sperm were distributed widely on various interfaces of the discontinuous Percoll density gradient (1.06 to 1.11 gm/ml). The content of Y-bearing sperm in the fraction lighter than 1.06 gm/ml was 73.1% +/- 3.3% and was reduced gradually along the density increment. Finally, it was found to be 27.4% +/- 3.4% in the sediment (more than 1.11 gm/ml).