Abstract
Those investigators who study the morphology of Legionella and Legionella-infected cells have greatly benefited from the superior resolution afforded by electron microscopy (EM). It can also be said with confidence that EM will continue to reveal as yet to be discovered features of this fascinating intracellular pathogen. In this chapter we detail our practical experience in the application of three transmission electron microscopy (TEM) techniques to the study of Legionella: conventional ultrastructural analysis, immuno-gold labeling, and negative staining. Each of these techniques has particular, well-defined applications, which are discussed in the context of our in-house developed methods. We invite researchers to try the methods given here in the study of Legionella, and adopt TEM as part of their research tools arsenal.
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References
Rodgers FG, Macrae AD, Lewis MJ (1978) Electron microscopy of the organism of Legionnaires’ disease. Nature 272:825–826
Rodgers FG (1979) Ultrastructure of Legionella pneumophila. J Clin Pathol 32:1195–1202
Gress FM, Myerowitz RL, Pascule AW, Rinaldo CR Jr, Dowling JN (1980) The ultrastructural morphologic features of Pittsburgh pneumonia agent. Am J Pathol 101:63–78
Horwitz MA (1983) Formation of a novel phagosome by the Legionnaires’ disease bacterium (Legionella pneumophila) in human monocytes. J Exp Med 158:1319–1331
Horwitz MA (1983) The Legionnaires’ disease bacterium (Legionella pneumophila) inhibits phagosome-lysosome fusion in human monocytes. J Exp Med 158:2108–2126
Garduño RA, Quinn F, Hoffman PS (1998) HeLa cells as a model to study the invasiveness and biology of Legionella pneumophila. Can J Microbiol 44:430–440
Faulkner G, Garduño RA (2002) Ultrastructural analysis of differentiation in Legionella pneumophila. J Bacteriol 184:7025–7041
Garduño RA, Garduño E, Hiltz M, Hoffman PS (2002) Intracellular growth of Legionella pneumophila gives rise to a differentiated form dissimilar to stationary phase forms. Infect Immun 70:6273–6283
Garduño RA, Faulkner G, Trevors MA, Vats N, Hoffman PS (1998) Immunolocalization of Hsp60 in Legionella pneumophila. J Bacteriol 180:505–513
Haniachi T, Sato T, Iwamoto T, Malavashi-Yamashiro Y, Hoshino M, Mizuno N (1986) A stable stain by modification of Sato’s method. J Electron Microsc 35:304–306
Molmeret M, Bitar DM, Han LH, Abu Kwaik Y (2004) Disruption of the phagosomal membrane and egress of Legionella pneumophila into the cytoplasm during the last stages of intracellular infection of macrophages and Acanthamoeba polyphaga. Infect Immun 72:4040–4051
Molmeret M, Marina S, Rexford A, Carabeo RA, Abu Kwaik Y (2007) Rapid escape of the dot/icm mutants of Legionella pneumophila into the cytosol of mammalian and protozoan cells. Infect Immun 75:3290–3304
Pease DC (1964) Histological techniques for electron microscopy. Academic, London
Hayat MA (1970) Principles and techniques of electron microscopy—biological applications, vol 1. Van Nostrand Reinhold Company, New York
Dawes CJ (1971) Biological techniques in electron microscopy. Barnes & Noble, New York
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Faulkner, G., Garduño, R.A. (2013). Electron Microscopy of Legionella and Legionella-Infected Cells. In: Buchrieser, C., Hilbi, H. (eds) Legionella. Methods in Molecular Biology, vol 954. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-161-5_17
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DOI: https://doi.org/10.1007/978-1-62703-161-5_17
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-62703-161-5
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