Optimizing sperm function: examining the correlation between sperm preparation techniques and membrane integrity plus capacitation dynamics

Background: Sperm preparation is a critical step in Assisted Reproductive Technology (ART) that significantly impacts sperm quality and fertilization potential. Membrane integrity and capacitation are the most significant parameters to assess sperm function, but methods like Swim-Up (SU) and Density Gradient Centrifugation (DGC) could influence these values by inducing mechanical damage. Methods: Ninety normozoospermic semen samples were obtained from ART patients and randomly divided into three groups (n=30 per group). Samples were processed by SU, DGC, or Microfluidics (MF) techniques. Initial motility and post-processing membrane integrity were evaluated according to routine protocols, including the Hypo-Osmotic Swelling Test. Statistical analysis was done with one-way ANOVA. Results: Initial motility was uniformly high across all groups (SU: 99%, DGC and MF: 100%). However, post-processing membrane integrity varied significantly (p<0.0001), with MF showing the highest integrity (86.03±1.98%), followed by SU (78.44±2.70%) and DGC (67.52±3.72%). Microscopic analysis corroborated these findings, indicating superior morphological preservation in the MF group. Conclusion: Microfluidics significantly outperforms traditional sperm preparation methods in preserving sperm membrane integrity at the cost of no motility compromise. Its adoption into ART protocols can potentially enhance sperm selection and deliver better treatment outcomes for fertility treatment.