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Quantitative Analysis of Phagocytosis and Phagosome Maturation

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Dictyostelium discoideum Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 983))

Abstract

Phagocytosis and phagosome maturation lead to killing and digestion of bacteria by protozoans and innate immune phagocytes. Phagocytosis of particles expressing or coupled to various fluorescent reporters and sensors can be used to monitor quantitatively various parameters of this central biological process. In this chapter we detail different labeling techniques of bacteria and latex beads used to measure adhesion and uptake by FACS analysis. We also describe methods to use fluorescent reporter dyes (FITC or DQgreen) coupled to silica beads to measure the kinetics of acidification and proteolysis. Measurements can be performed either at the single-cell level, using live microscopy, or for a whole cell population, with a fluorescence microplate reader.

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Acknowledgements

We gratefully acknowledge Rashid A. Mohamed for preliminary experiments with the phagocytosis assay. The laboratory of Thierry Soldati is supported by multiple grants from the Swiss National Science Foundation.

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Authors and Affiliations

  1. Départment de Biochimie, Faculté des Sciences, Université de Genève, Geneva, Switzerland

    Natascha Sattler & Thierry Soldati

  2. Départment de Biochimie, Faculté des Sciences, Université de Genève, Genève, Switzerland

    Roger Monroy

Authors
  1. Natascha Sattler
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  2. Roger Monroy
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  3. Thierry Soldati
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Corresponding author

Correspondence to Thierry Soldati .

Editor information

Editors and Affiliations

  1. , Centre for Biochemistry, University of Cologne, Joseph-Stelzmann-Str. 52, Köln, 50931, Nordrhein-Westfalen, Germany

    Ludwig Eichinger

  2. , Centre for Biomedical Research, University of Hull, Hull, HU6 7RX, Montserrat

    Francisco Rivero

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Sattler, N., Monroy, R., Soldati, T. (2013). Quantitative Analysis of Phagocytosis and Phagosome Maturation. In: Eichinger, L., Rivero, F. (eds) Dictyostelium discoideum Protocols. Methods in Molecular Biology, vol 983. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-302-2_21

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  • DOI: https://doi.org/10.1007/978-1-62703-302-2_21

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-301-5

  • Online ISBN: 978-1-62703-302-2

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