Analysis of cytokinesis by electron microscopy

Methods Cell Biol. 2017:137:225-238. doi: 10.1016/bs.mcb.2016.03.029. Epub 2016 Jun 28.

Abstract

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.

Keywords: Abscission; Correlative light electron microscopy; Cytokinesis; Electron microscopy; Electron tomography; High-pressure freezing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Caenorhabditis elegans / ultrastructure
  • Cytokinesis / genetics*
  • Electron Microscope Tomography / methods*
  • Embryo, Nonmammalian / ultrastructure*
  • Embryonic Development / genetics
  • Microscopy, Electron / methods*
  • Mitosis / genetics
  • Sea Urchins / genetics
  • Sea Urchins / growth & development
  • Sea Urchins / ultrastructure