The use of pentoxifylline in boosting the sorting efficacy of low viability mammalian sperm via micro fluidic sperm bio-chip

Traditional sperm sorting methods all requires manual selection, which is tedious and often cause sperm damage during the screen process. Although recent advances in microfluidic sperm sorter design, seemed to point for a possibility to make possible the separation of sperms into various mobility subclasses. However, under cytohistological observation the sorted live sperm from these devices, as is corresponded with more diffused blue stains, was often few in number and not as motile. Therefore, in order to confirm the real sorting efficacy of MISS, in this study the amount of sperm collected was counted by flow cytometry and attempt was made to establish the statistic model. Results from t-test with 95% confidence level shown a significant difference in live/dead sperm ratio. Also, from the combined result among flow cytometry, fluorescent stain, and the statistic model test, it is confirmed that live sperms were able to cross the laminar stream created by the device and reach the sorting chamber with corresponding range from non-motile sperms to debris, and from living to dead ones. It is thus concluded that the proposed method is cost effective and is possible to serve as a protocol for rapid sperm quantitative assessment.