Ultrastructural investigation methods for Trypanosoma brucei.
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.
Höög
Johanna L
JL
Sir William Dunn School of Pathology, University of Oxford, Oxford OX13RE, United Kingdom.
Gluenz
Eva
E
Vaughan
Sue
S
Gull
Keith
K
D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques; D003599:Cytoskeleton / Q000648:ultrastructure; D017110:Freeze Substitution / Q000379:methods; D006652:Histological Techniques / Q000379:methods*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D046529:Microscopy, Electron, Transmission / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods; D014346:Trypanosoma brucei brucei / Q000648:ultrastructure*