Scanning and three-dimensional electron microscopy methods for the study of Trypanosoma brucei and Leishmania mexicana flagella

Gluenz, E. , Wheeler, R. J., Hughes, L. and Vaughan, S. (2015) Scanning and three-dimensional electron microscopy methods for the study of Trypanosoma brucei and Leishmania mexicana flagella. In: Methods in Cilia and Flagella. Series: Methods in cell biology, 127. Elsevier, pp. 509-542. (doi:10.1016/bs.mcb.2014.12.011)

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Abstract

Three-dimensional electron microscopy tools have revolutionized our understanding of cell structure and molecular complexes in biology. Here, we describe methods for studying flagellar ultrastructure and biogenesis in two unicellular parasites—Trypanosoma brucei and Leishmania mexicana. We describe methods for the preparation of these parasites for scanning electron microscopy cellular electron tomography, and serial block face scanning electron microscopy (SBFSEM). These parasites have a highly ordered cell shape and form, with a defined positioning of internal cytoskeletal structures and organelles. We show how knowledge of these can be used to dissect cell cycles in both parasites and identify the old flagellum from the new in T. brucei. Finally, we demonstrate the use of SBFSEM three-dimensional models for analysis of individual whole cells, demonstrating the excellent potential this technique has for future studies of mutant cell lines.

Item Type:Book Sections
Status:Published
Glasgow Author(s) Enlighten ID:Gluenz, Dr Eva
Authors: Gluenz, E., Wheeler, R. J., Hughes, L., and Vaughan, S.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Publisher:Elsevier
ISSN:0091-679X

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