Cell-free extract systems and the cytoskeleton: preparation of biochemical experiments for transmission electron microscopy

Methods Mol Biol. 2007;369:199-212. doi: 10.1007/978-1-59745-294-6_10.

Abstract

Cell-free systems can be used to reconstitute complex actin or microtubule-based phenomena. For example, a number of extracts support actin-dependent propulsion of Listeria monocytogenes, whereas Xenopus laevis extracts support formation of a microtubule-based meiotic spindle. Working in vitro opens these complex processes to biochemical dissection. Here, we describe methods to view these in vitro preparations by thin-section electron microscopy.

MeSH terms

  • Actins / metabolism
  • Animals
  • Biotin
  • Cell-Free System
  • Cytoskeleton / metabolism*
  • Cytoskeleton / ultrastructure*
  • Female
  • Fixatives
  • Listeria monocytogenes / metabolism
  • Listeria monocytogenes / ultrastructure
  • Microscopy, Electron, Transmission / methods*
  • Ovum / metabolism
  • Ovum / ultrastructure
  • Plastic Embedding
  • Spindle Apparatus / metabolism
  • Spindle Apparatus / ultrastructure
  • Streptavidin
  • Xenopus laevis

Substances

  • Actins
  • Fixatives
  • Biotin
  • Streptavidin