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Live Imaging of Mycobacterium marinum Infection in Dictyostelium discoideum

  • Caroline Barisch
  • Ana T. López-Jiménez
  • Thierry SoldatiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1285)

Abstract

The Dictyostelium discoideumMycobacterium marinum host–pathogen system is a recently established and powerful model system for mycobacterial infection. In this chapter, two simple protocols for live imaging of Dictyostelium discoideum infection are described. The first method is used to monitor the dynamics of recruitment of GFP-tagged Dictyostelium discoideum proteins at single time-points corresponding to the main stages of the infection (1.5–72 h post infection). The second method focuses at the early stages of the establishment of an infection (0–3 h post infection). In addition, several procedures to improve the imaging of the bacterium-containing compartment are described. Basic bacterial parameters such as bacterial growth and the recruitment of host proteins to the bacterium-containing compartment can be easily and precisely quantified using macros for ImageJ. These methods can be adapted to monitoring mycobacteria infection in other systems using mammalian cells.

Key words

Dictyostelium discoideum Mycobacteria Intracellular infection Live microscopy Fluorescence 

Notes

Acknowledgements

The TS laboratory is supported by multiple grants from the Swiss National Science Foundation.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Caroline Barisch
    • 1
  • Ana T. López-Jiménez
    • 1
  • Thierry Soldati
    • 1
    Email author
  1. 1.Département de Biochimie, Faculté des SciencesUniversité de Genève, Sciences IIGenève-4Switzerland

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