Author:
Date:
2020-10-20
[Abstract] Live-imaging of meiotic cell division has been performed in extracted spermatocytes of a number of species using phase-contrast microscopy. For the nematode Caenorhabditis elegans, removal of spermatocytes from gonads has damaging effects, as most of the extracted spermatocytes show a high variability in the timing of meiotic divisions or simply arrest during the experiment. Therefore, we developed a live-cell imaging approach for in situ filming of spermatocyte meiosis in whole immobilized C. elegans males, thus allowing an observation of male germ cells within an ...
[摘要] [摘要] 用相差显微镜对一些物种的精母细胞进行了减数分裂的实时成像。对于秀丽隐杆线虫来说,去除生殖腺中的精母细胞具有破坏性作用,因为大多数精母细胞在减数分裂的时间上表现出高度的变异性,或者只是在实验中停止。因此,我们开发了一种活体细胞成像方法,用于原位拍摄固定化线虫雄性精母细胞减数分裂过程,从而可以在不受干扰的环境中观察雄性生殖细胞。为此,我们利用带有荧光标记染色体和中心体的菌株。在这里我们描述如何固定男性蠕虫进行实时成像。此外,我们描述了获取和处理数据的工作流程,以获得有关精母细胞减数分裂I和II中染色体分离动态的定量信息。此外,我们最新开发的方法允许我们在电子显微镜中重新定位胶片上的主轴,而不管蜗杆的初始3D方位如何,并以统计稳健的方式分析活蠕虫的纺锤动力学。我们的实时成像方法也适用于秀丽隐杆线虫雌雄同体,并且可以扩展到其他荧光标记的线虫或其他完全透明的小型模型生物。 [背景] ...
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