Comparison of the external physical damages between laser-assisted and mechanical immobilized human sperm using scanning electronic microscopy
Comparison of the external physical damages between laser-assisted and mechanical immobilized human sperm using scanning electronic microscopy
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DOI：
10.1371/journal.pone.0188504
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发表时间：
2017-11-27
期刊：
影响因子：
3.7
通讯作者：
Li, Tin Chiu
中科院分区：
文献类型：
--
作者：
Chan, David Y. L.;Li, Tin Chiu
我们的目的是可视化的外部物理损伤和不同的外部表型效应之间的机械和激光辅助固定人类精子使用扫描电子显微镜（SEM）。用机械法和激光辅助法固定人精子进行扫描电镜观察，并对两种固定精子的膜完整性进行了检查。我们发现在SEM水平上机械扭结精子的外部损伤的证据，但不是在激光辅助固定精子。虽然激光辅助固定精子没有发现外部损伤，但当精子被红外激光照射时，有两种不同类型的形态学变化。当激光脉冲应用于精子末端区域时，立即形成卷曲的尾部，而激光应用于精子主片区域导致精子尾部急剧弯曲。如果激光没有击中屏幕上的中央目标区域，或者如果激光击中精子中段或头部，则激光固定的精子不会表现出任何形态学变化。我们改良的膜完整性检测显示，超过一半的激光辅助固定精子的外膜保持完整。总之，机械固定产生膜损伤，而激光辅助固定除了在SEM水平上的形态学变化外，没有导致任何外部膜损伤。
We aim to visualize the external physical damages and distinct external phenotypic effects between mechanical and laser-assisted immobilized human spermatozoa using scanning electronic microscopy (SEM). Human spermatozoa were immobilized mechanically or with laser assistance for SEM examination and the membrane integrities were checked on both types of immobilized spermatozoa. We found evidence of external damages at SEM level on mechanically kinked sperm, but not on laser-assisted immobilized sperm. Although no external damage was found on laser-assist immobilized sperm, there were two distinct types of morphological changes when spermatozoa were stricken by infra-red laser. Coiled tails were immediately formed when Laser pulse was applied to the sperm end piece area, whereas laser applied to the sperm principal piece area resulted in a sharp bend of sperm tails. Sperm immobilized by laser did not exhibit any morphological change if the laser did not hit within the on-screen central target zone or if the laser hit the sperm mid piece or head. Our modified membrane integrity assay revealed that the external membrane of more than half of the laser-assisted immobilized sperm remained intact. In conclusion, mechanical immobilization produced membrane damages whilst laser-assisted immobilization did not result in any external membrane damages besides morphological changes at SEM level.