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Analysis of Legionella Infection by Flow Cytometry

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Legionella

Part of the book series: Methods in Molecular Biology ((MIMB,volume 954))

Abstract

Legionella pneumophila infects and replicates in environmental protozoa and metazoan macrophages within a specific vacuole. The infection of phagocytes by L. pneumophila can be assessed by an agar plating assay or by fluorescence microscopy. Here, we describe the analysis of Legionella infection by automated flow cytometry using wild-type and mutant bacteria that constitutively produce the green fluorescent protein (GFP). Advantages of the flow cytometry technique include (1) a software-assisted multiple parameter analysis of Legionella infections in real-time at distinct stages of the infection cycle, (2) the simultaneous and fast acquisition of a high number of data points, and (3) a characterization of the infecting bacteria in parallel with the infected host cells.

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Abbreviations

ACES:

N-(2-Acetamido)-2-aminoethanesulfonic acid

icm/dot :

Intracellular multiplication/defective organelle trafficking

MES:

2-N-Morpholinoethanesulfonic

PI:

Propidium iodide

T4SS:

Type IV secretion system

FSC:

Forward scatter channel

SSC:

Sideward scatter channel

GFP:

Green fluorescent protein

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Acknowledgments

This work was supported by the Max von Pettenkofer Institute, Ludwig-Maximilians University Munich, and the German Research Foundation (HI 1511/2-1).

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Authors and Affiliations

  1. Center for Applied Biotechnology and Molecular Medicine, University of Zurich, Zurich, Switzerland

    André N. Tiaden

  2. Max von Pettenkofer Institute, Ludwig-Maximilians University, Munich, Germany

    Aline Kessler & Hubert Hilbi

Authors
  1. André N. Tiaden
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  2. Aline Kessler
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  3. Hubert Hilbi
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Corresponding author

Correspondence to Hubert Hilbi .

Editor information

Editors and Affiliations

  1. , Biology of Intracellular Bacteria, Institut Pasteur, Rue du Dr. Roux 28, Paris, 75724, France

    Carmen Buchrieser

  2. Max von Pettenkofer Institute, Ludwig-Maximilians-University, Pettenkoferstrasse 9a, Munich, 80336, Germany

    Hubert Hilbi

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Tiaden, A.N., Kessler, A., Hilbi, H. (2013). Analysis of Legionella Infection by Flow Cytometry. 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_14

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  • DOI: https://doi.org/10.1007/978-1-62703-161-5_14

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-160-8

  • Online ISBN: 978-1-62703-161-5

  • eBook Packages: Springer Protocols

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