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JoVE Journal
Immunology and Infection

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JoVE Journal Immunology and Infection
Single Cell Measurements of Vacuolar Rupture Caused by Intracellular Pathogens
 

Single Cell Measurements of Vacuolar Rupture Caused by Intracellular Pathogens

Article DOI: 10.3791/50116-v • 10:39 min • June 12th, 2013
June 12th, 2013 •
*1, *1, 2, 3, 3, 1, 1
1Dynamique des Interactions Hôte Pathogène, Institut Pasteur, Paris, France, 2Imagopole, Institut Pasteur, Paris, France, 3Pathogenomique Mycobacterienne Integrée, Institut Pasteur, Paris, France
* These authors contributed equally

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résumé
Automatic Translation

We describe a method for tracking the endomembrane rupture elicited by the intracellular bacteria Shigella flexneri and Mycobacterium tuberculosis upon host cell invasion. Our assay makes use of CCF4, a host cytoplasmic FRET probe in live or fixed cells. This reporter is degraded by an enzyme activity present on the bacterial surface.

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Keywords: Single Cell Vacuolar Rupture Intracellular Pathogens Shigella Flexneri Mycobacterium Tuberculosis CCF4 FRET Reporter Phagosomal Membrane Disruption Real-time Imaging High-throughput Analysis
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