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Applications of Microscopy in Salmonella Research

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Salmonella

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1225))

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Abstract

Salmonella enterica is a Gram-negative enteropathogen that can cause localized infections, typically resulting in gastroenteritis, or systemic infection, e.g., typhoid fever, in humans and many other animals. Understanding the mechanisms by which Salmonella induces disease has been the focus of intensive research. This has revealed that Salmonella invasion requires dynamic cross-talk between the microbe and host cells, in which bacterial adherence rapidly leads to a complex sequence of cellular responses initiated by proteins translocated into the host cell by a type 3 secretion system. Once these Salmonella-induced responses have resulted in bacterial invasion, proteins translocated by a second type 3 secretion system initiate further modulation of cellular activities to enable survival and replication of the invading pathogen. Elucidation of the complex and highly dynamic pathogen–host interactions ultimately requires analysis at the level of single cells and single infection events. To achieve this goal, researchers have applied a diverse range of microscopy techniques to analyze Salmonella infection in models ranging from whole animal to isolated cells and simple eukaryotic organisms. For example, electron microscopy and high-resolution light microscopy techniques such as confocal microscopy can reveal the precise location of Salmonella and its relationship to cellular components. Widefield light microscopy is a simpler approach with which to study the interaction of bacteria with host cells and often has advantages for live cell imaging, enabling detailed analysis of the dynamics of infection and cellular responses. Here we review the use of imaging techniques in Salmonella research and compare the capabilities of different classes of microscope to address specific types of research question. We also provide protocols and notes on some microscopy techniques used routinely in our own research.

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Acknowledgements

We would like to thank Alan Leard, Katy Jepson, Tom MacVicar, Joe Beesley, and Jon Jennings for assistance with microscopy, image processing, and protocols. We also acknowledge all our colleagues who have contributed to the studies of Salmonella infection and the development of microscopical techniques discussed here. Work in this laboratory as described here was supported by MRC, BBSRC, and Unilever. The University of Bristol Bioimaging Facility was initially established with funding from MRC and its subsequent expansion has been funded by The Wolfson Foundation, University of Bristol and MRC.

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  1. Department of Biochemistry, School of Medical Sciences, University of Bristol, Tyndall Avenue, Bristol, BS8 1TD, UK

    Layla M. Malt, Charlotte A. Perrett, Suzanne Humphrey & Mark A. Jepson

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  1. Layla M. Malt
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  2. Charlotte A. Perrett
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  3. Suzanne Humphrey
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  4. Mark A. Jepson
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Correspondence to Mark A. Jepson .

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  1. Department of Veterinary Pathobiology, University of Missouri, Columbia, Missouri, USA

    Heide Schatten

  2. Cancer Research Center, University of Missouri, Columbia, Missouri, USA

    Abraham Eisenstark

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Malt, L.M., Perrett, C.A., Humphrey, S., Jepson, M.A. (2015). Applications of Microscopy in Salmonella Research. In: Schatten, H., Eisenstark, A. (eds) Salmonella. Methods in Molecular Biology, vol 1225. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1625-2_12

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  • DOI: https://doi.org/10.1007/978-1-4939-1625-2_12

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1624-5

  • Online ISBN: 978-1-4939-1625-2

  • eBook Packages: Springer Protocols

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