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Separation of Progressive Motile Sperm from Mouse Semen Using On-chip Chemotaxis

机译:使用片上趋化性从小鼠精液中分离进行性运动精子

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We present a novel method for the separation of progressive motile sperm from non-progressive motile and immotile sperm. This separation was accomplished by inducing chemotaxis along a longitudinal chemical gradient in a microchip composed of a biocompatible polydimethysiloxane layer and a glass substrate. In a preliminary experiment using fluorescent rhodamine B as a marker, we verified that a chemical gradient was generated by diffusion within the microchannel. We used acetylcholine as a chemoattractant to evaluate the chemotactic response of sperm. We tested the response to a 1/2 to 1/64 dilution series of acetylcholine. The results of a mouse sperm chemotaxis assay showed that progressive motile sperm swam predominantly toward the outlet at an optimal chemical gradient of 0.625 (mg/ml)/mm of acetylcholine. This device provides a convenient, disposable, and high-throughput platform that could function as a progressive motile sperm sorter for potential use in intracytoplasmic sperm injection.
机译:我们提出了从非进行性运动和不运动性精子中分离进行性运动精子的新方法。通过在由生物相容性聚二甲基硅氧烷层和玻璃基板组成的微芯片中沿纵向化学梯度诱导趋化作用来实现这种分离。在使用荧光若丹明B作为标记的初步实验中,我们验证了通过微通道内扩散产生的化学梯度。我们使用乙酰胆碱作为趋化剂来评估精子的趋化反应。我们测试了对1/2至1/64稀释的乙酰胆碱系列的反应。小鼠精子趋化性试验的结果表明,进行性运动精子主要以最佳化学梯度0.625(mg / ml)/ mm乙酰胆碱游向出口。该设备提供了一个方便,可抛弃的高通量平台,可作为进行性运动型精子分选仪,潜在地用于胞浆内精子注射。

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