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Bacterial Virulence pp 105–117Cite as

Flow Cytometry–Based Single Cell Analyses of Bacterial Adaptation to Intracellular Environments

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Part of the Methods in Molecular Biology book series (MIMB,volume 2427)

Abstract

Since decades, flow cytometry (FC) is a powerful technique to perform single cell analyses with high accuracy and throughput. Moreover, FC is the method of choice to study bacterial cell heterogeneity and complements single-cell imaging techniques. The complex experimental approaches for FC sample preparation and the subsequent FC adjustment and gating strategy demand careful considerations to be successful when analyzing complex microbial populations, especially when liberated populations of intracellular bacterial pathogens, or bacterial pathogens inside intact host cells are analyzed. Here, we provide a set of experimental protocols for FC sample preparation of (1) in vitro cultured bacterial cells, (2) liberated intracellular bacteria from host cells, or (3) preparation of intact infected phagocytic or epithelial cells commonly used as host cells in infection biology. Since sample preparation, cytometer adjustment, and gating strategy are essential for experimental success, we aim to provide our expertise to support application of FC by other researchers.

Key words

  • Flow cytometry
  • Single cell analysis
  • Intracellular bacteria
  • Host cells
  • High-throughput analyses

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  • DOI: 10.1007/978-1-0716-1971-1_10
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Acknowledgments

This work was supported by the DFG by project P15 in SFB 944.

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Authors and Affiliations

  1. Abteilung Mikrobiologie and CellNanOs—Center of Cellular Nanoanalytics Osnabrück, Fachbereich Biologie/Chemie, Universität Osnabrück Barbarastr, Osnabrück, Germany

    Marc Schulte & Michael Hensel

Authors
  1. Marc Schulte
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  2. Michael Hensel
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Corresponding author

Correspondence to Michael Hensel .

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Editors and Affiliations

  1. The Infectious Diseases Research Laboratory, Sheba Medical Center, Tel-Hashomer, Israel

    Prof. Ohad Gal-Mor

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Schulte, M., Hensel, M. (2022). Flow Cytometry–Based Single Cell Analyses of Bacterial Adaptation to Intracellular Environments. In: Gal-Mor, O. (eds) Bacterial Virulence. Methods in Molecular Biology, vol 2427. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1971-1_10

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  • DOI: https://doi.org/10.1007/978-1-0716-1971-1_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1970-4

  • Online ISBN: 978-1-0716-1971-1

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