Quantification of X sperm by raman spectroscopy in percoll density gradient centrifuged buffalo semen

The present study was conducted to observe effect of percoll density gradient centrifugation of buffalo bull semen on quantity of X sperms. Ejaculates were collected by artificial vagina method. Semen with mass motility >+3 and progressive motility >70 % were selected for experiment. X sperm Enrichment of semen was done by discontinuous percoll density gradient centrifugation and three groups were formed ie Group 1  (3 layer 70%, 50% and 30%) Group 2  (7 layer 70%, 60%, 50%, 40%, 30%, 20% and 10%) Group 3  (7 layer 80%, 70%, 60%, 50%, 40%, 30% and 20%). Centrifugation of semen of three groups and control (fresh semen without gradients) was done. After centrifugation, the supernatant part was removed and the pellet of each group was used for X sperm enrichment assessment by Raman spectroscopy. Results revealed that X sperm enrichment was higher in the pellets of Group 2 followed Group 3, Group 1 and Control as Raman peaks on DNA specific bands corresponds to more number of x sperm were higher respectively.