Ultrastructural investigation methods for Trypanosoma brucei

Methods Cell Biol. 2010;96:175-96. doi: 10.1016/S0091-679X(10)96008-1.

Abstract

Trypanosoma brucei is a unicellular parasite causing African sleeping sickness in cattle and humans. Due to the ease with which these cells can be cultured and genetically manipulated, it has emerged as a model organism for the kinetoplastids.In this chapter we describe the preparation of T. brucei for transmission electron microscopy. A thorough explanation of conventional sample preparation through chemical fixation of whole cells and detergent extracted cytoskeletons followed by dehydration and Epon embedding is given. We also introduce a novel high-pressure freezing protocol, which followed by rapid freeze substitution and HM20 embedding generates T. brucei samples displaying good cell morphology, which are suitable for immunocytochemistry.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Cell Culture Techniques
  • Cytoskeleton / ultrastructure
  • Freeze Substitution / methods
  • Histological Techniques / methods*
  • Humans
  • Immunohistochemistry / methods*
  • Microscopy, Electron, Transmission / methods*
  • Staining and Labeling / methods
  • Trypanosoma brucei brucei / ultrastructure*