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

English

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13,305 Views

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10:39 min

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June 12, 2013

DOI:

10.3791/50116-v

DOI:
10.3791/50116-v

•

10:39 min
June 12, 2013

•

1 Views
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1Dynamique des Interactions Hôte Pathogène,Institut Pasteur, Paris, France, 2Imagopole,Institut Pasteur, Paris, France, 3Pathogenomique Mycobacterienne Integrée,Institut Pasteur, Paris, France

Summary

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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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Single CellVacuolar RuptureIntracellular PathogensShigella FlexneriMycobacterium TuberculosisCCF4 FRET ReporterPhagosomal Membrane DisruptionReal-time ImagingHigh-throughput Analysis

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Cite this Article

Keller, C., Mellouk, N., Danckaert, A., Simeone, R., Brosch, R., Enninga, J., Bobard, A. Single Cell Measurements of Vacuolar Rupture Caused by Intracellular Pathogens. J. Vis. Exp. (76), e50116, doi:10.3791/50116 (2013).

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