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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.

Original publication

DOI

10.1016/S0091-679X(10)96008-1

Type

Journal article

Journal

Methods Cell Biol

Publication Date

2010

Volume

96

Pages

175 - 196

Keywords

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