Micro-Raman spectroscopy reveals distinct differences in sperm quality selected by hyaluronic acid

While the association between sperm selection with Hyaluronic Acid (HA) and DNA quality is well-accepted, the underlying mechanism remains unclear. In order to shed light on this issue, micro-Raman spectroscopy was utilized to analyze the differences in the Raman spectral response of sperm cells bound with or without HA. Our results demonstrate that the Raman spectra of HA-bound sperm and HA-unbound sperm display distinct differences in Raman spectral response. Furthermore, we conducted nucleoprotein and nuclear DNA fluorescent staining to validate our findings, demonstrating that HAbound sperm exhibit enhanced maturity in nucleoproteins and heightened DNA duplex quality. These findings substantiate the effectiveness of HA in selectively identifying highquality sperm cells. In summary, our research suggests that HA can be a valuable component for efficiently selecting high-quality sperm cells, and the integration of HA screening with micro-Raman spectroscopy presents a promising, non-invasive approach for objectively evaluating and screening strictly high-quality such cells.