Received April 08, 2024
Accepted October 08, 2024

Objective: Sperm parameters and DNA integrity are crucial factors in ART outcomes. This study compared four sperm preparation methods (microfluidics, MACS, zeta potential, and swim-Up) for sorting spermatozoa with normal parameters and chromatin integrity. Methods: This study evaluated semen samples from 25 couples with male factor infertility. The semen samples were divided into four portions: one prepared by MACS, one by zeta potential, the other by microfluidics, and the last by swim-up. After preparation, sperm viability, motility, and morphology were assessed based on the WHO guidelines. DNA intergrity was assessed by SDF assay, and the CMA3 staining test was used to evaluate sperm chromatin packaging. Results: Compared to other preparation techniques, microfluidic preparation significantly improved sperm parameters, including motility, viability, morphology, and DNA integrity as well as chromatin packaging (P-value <0.05). The results also demonstrated that sperm motility, viability, and sperm DNA integrity as well as chromatin packaging, were not significantly different after preparation with MACS and Zeta potential methods. However, the MACS and Zeta methods produced improved sperm parameters and better DNA integrity than the swim-up method. Conclusions: Our results indicate that microfluidics can improve sperm quality compared to other methods of sperm preparation. When the microfluidic chip is not available, considering the similar results of sperm preparation by MACS and Zeta potential methods, it is preferred to use the Zeta method for the ART cycle due to its simplicity and cost-effectiveness.