Reichmann Judith, Eguren Manuel, Lin Yu, Schneider Isabell, Ellenberg Jan
European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
Methods Cell Biol. 2018;145:279-292. doi: 10.1016/bs.mcb.2018.03.030. Epub 2018 Apr 26.
Systematic studies of cell divisions at the beginning of mammalian life are of fundamental importance for our understanding of embryonic development and fertility. However, in the past the challenges of in vitro embryo culture and the embryo's pronounced light sensitivity have precluded a detailed investigation of preimplantation cell divisions. This protocol is based on recent technological breakthroughs in inverted light microscopy tailored for mouse embryology. Due to its reduced light dose, and therefore low phototoxicity, as well as higher acquisition speed, light-sheet microscopy allows extended 3D time-lapse imaging of early embryonic development with very high spatial and temporal resolution. This imaging approach enables imaging of key subcellular structures during the critical cell cycles from the zygote up to the blastocyst stage, with a resolution that allows automatic computational tracking and quantitative analysis of the dynamics of mitotic organelles.
对哺乳动物生命起始阶段细胞分裂进行系统研究，对于我们理解胚胎发育和生育能力至关重要。然而，过去体外胚胎培养面临的挑战以及胚胎对光的显著敏感性，使得对植入前细胞分裂的详细研究受到阻碍。本方案基于为小鼠胚胎学量身定制的倒置光学显微镜的最新技术突破。由于光片显微镜的光剂量降低，因此光毒性低，以及采集速度更高，它能够以非常高的空间和时间分辨率对早期胚胎发育进行长时间的三维延时成像。这种成像方法能够在从合子到囊胚阶段的关键细胞周期中对关键亚细胞结构进行成像，其分辨率足以实现对有丝分裂细胞器动态的自动计算跟踪和定量分析。
