Microfluidic systems with the ability to mimic the female reproductive tract (FRT) and sperm features have emerged as promising methods to separate sperm with higher quality for the assistant reproductive technology. Thereby, we designed and fabricated a microfluidic system based on FRT features with a focus on rheotaxis and thigmotaxis for passive sperm separation. In this regard, four various geometries (linear, square, zigzag, and sinusoidal) were designed, and the effect of rheotaxis and thigmotaxis were investigated. Although separated sperm in all microchannels were 100% motile, non-linear geometries were more effective than linear geometry in the term of separating the progressive sperm with high quality. In the presence of upstream flow, periodical changes in the slope of walls (in non-linear geometries) give rise to the periodical facing sperm with a high flow rate in the middle of microchannels, which was a reason for the high quality of separated sperm. However, because of sharp corners in the square and zigzag microchannels that create dead zones with a lack of upstream flow, which is noticeable via simulation results, these geometries have obstacles against sperm swimming toward the outlet, which was proved by image analysis. The sinusoidal geometry showed the highest enhancement level of the designed geometries compared to the linear geometry. Separated sperm exhibited 34.7% normal morphology, 100% motility, and 100% viability in the sinusoidal geometry. Therefore, the periodic change in the position of sperm from one wall to another wall can be a strategy for separating sperm with high quality.
Graphical abstract: In the present study, we used a microfluidic system for studying the combined effects of thigmotaxis and rheotaxis for sperm separation process to achieve the successful Assisted reproductive technology (ART). The designed PDMS-based microfluidic system had four various geometries, including linear, square, zigzag, and sinusoidal. The functionality of separated sperm was evaluated by sperm tracking (ImageJ), motility assay (CASA software), and morphology assay (Papanicolaou ultrafast staining). Probing various geometries revealed 100% motility. In non-linear geometries, sperm's periodic detachment from the walls gave rise to the periodic interaction with the high flow velocity in the center of the channel, resulting in the separation of high-quality sperm with progressive motility. The collected data proved the influence of thigmotaxis on the quality of separated sperm. Morphologically improvement in separated sperm from the sinusoidal geometry was significant than others, which means the sinusoidal structure would be the best candidate for the sperm separation process.
Supplementary information: The online version contains supplementary material available at 10.1007/s13534-023-00294-8.
Keywords: Female reproductive tract; Microfluidic system; Rheotaxis; Sperm separation; Thigmotaxis.
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