König J, Borrego-Pinto J, Streichert D, Munzig M, Lenart P, Müller-Reichert T
Technische Universität Dresden, Dresden, Germany.
European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
Methods Cell Biol. 2017;137:225-238. doi: 10.1016/bs.mcb.2016.03.029. Epub 2016 Jun 28.
Following up on a chapter on the Correlative Light and Electron Microscopy of Early Caenorhabditis elegans Embryos in Mitosis (MCB 79, 101-119), we present an adaptation of our established protocol for the ultrastructural analysis of either permeabilized or injected embryonic systems. We prepared both drug-treated early C. elegans embryos and fluorescently labeled sea urchin embryos of Lytechinus pictus for ultrastructural studies on animal cytokinesis. Here we focus on the initial preparation steps of postmitotic embryos for high-pressure freezing and subsequent electron microscopy with an emphasis on electron tomography. The advantages and limitations of our extended protocol will be discussed.
继《秀丽隐杆线虫有丝分裂早期胚胎的相关光镜和电镜观察》一章（《分子与细胞生物学》79卷，第101 - 119页）之后，我们展示了对已建立的用于通透化或注射胚胎系统超微结构分析方案的一种改进。我们制备了经药物处理的秀丽隐杆线虫早期胚胎以及荧光标记的刺冠海胆胚胎，用于动物胞质分裂的超微结构研究。在此，我们重点关注有丝分裂后胚胎用于高压冷冻及后续电子显微镜观察的初始制备步骤，尤其强调电子断层扫描。我们还将讨论扩展方案的优缺点。
