Zebrafish sperm activation is a necessary step in zebrafish research to satisfy large demands for current vertebral experiments, and the success of sperm motility is significantly affected by the magnitude of fluid-induced shear force. In traditional methods, zebrafish sperms are activated by rapidly diluting them in an aqueous medium through manual mixing. However, this method is inefficient because of low activation rates. This study is aimed to develop an artificial cilia-based micromixer through the precise regulation of hydrodynamics induced on zebrafish sperms for superior sperm activation. Results revealed that the proposed device activated as high as 68% of the tested sperms compared with an average of 44% sperms activated through the manual mixing. Therefore, sperm activation was significantly improved by 50% (p < 0.05). Parameters such as artificial rotation frequency and artificial cilia actuation time period were further evaluated and characterized. Zebrafish sperms were found to be activated within 5 s by using the proposed device. Thus, the proposed micromixer is envisaged that can advance the studies related to sperm dynamics and in-vitro fertilization procedures for vertebrate.
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