Integration of Micro Fluidic Bio-chip Design and Automatic fluorescent Identification for Rapid Sperm Mobility Assessment

Abstract

Since male abnormality has now been reported to account at least thirty percent of couples with fertility problem. Therefore, distinguishing and classification of sperm has become more and more important, especially when Assisted Reproductive Technology (ART) like in vitro fertilization (IVF) is involved. However, traditional sperm sorting methods all requires manual selection, which is tedious and often cause sperm damage during the screen process. Although, new design from our recent study, seemed to point for a possibility to make these kind of MISS device capable of separating sperms into various mobility sub-class. In addition, the Hoechst 33258 and propidium iodide double staining results, also confirmed that the most motile and health sperm is indeed able to leave the laminar stream and being sorted out at outlet-h, with least dead sperm or debris. Meanwhile, we observed the dead sperm had been dominate from clear red stains and live sperm are not as active as is corresponded with more diffused blue stains. In order to confirm the real sorting situation, we counted the amount of sperm in collected sample by flow cytometry and tried to establish the statistic model. Though the flow cytometry, we could detect the sperm fluorescent distribution and count the size of sperm population. Nevertheless, t-test with 95% confidence level also shown significant difference in live/dead sperm ratio. Therefore, 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, as well as from living to dead ones. It is concluded that the proposed method is cost effective and is possible to serve as a protocol for rapid sperm quantitative assessment.