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

Ultrastructural investigation methods for Trypanosoma brucei.

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1
Sir William Dunn School of Pathology, University of Oxford, Oxford OX13RE, United Kingdom.

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.

PMID:
20869523
DOI:
10.1016/S0091-679X(10)96008-1
[Indexed for MEDLINE]
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