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Journal / Immunology and Infection / Single Cell Measurements of Vacuolar Rupture…
JoVE Journal
Immunology and Infection

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Single Cell Measurements of Vacuolar Rupture Caused by Intracellular Pathogens
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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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Summary
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.

Tags

Single Cell Measurements Vacuolar Rupture Intracellular Pathogens Shigella Flexneri Mycobacterium Tuberculosis Endocytic Vacuole Phagosomal Membrane Robust Assay Tracking Phagosomal Membrane Disruption Host Cell Entry CCF4 FRET Reporter β-lactamase Cytosol Membrane-enclosed Compartments Vacuole Disruption Dynamics Automated Microscopy Image Processing
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