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Imunologia e Infecção
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Single Cell Measurements of Vacuolar Rupture Caused by Intracellular Pathogens
JoVE Journal
Imunologia e Infecção
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JoVE Journal Imunologia e Infecção
Single Cell Measurements of Vacuolar Rupture Caused by Intracellular Pathogens

Single Cell Measurements of Vacuolar Rupture Caused by Intracellular Pathogens

DOI:
10.3791/50116-v

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10:39 min

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June 12, 2013

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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

Capítulos

  • 00:05Título
  • 01:29CCF4 Assay on Fixed Samples Using Shigella flexneri (96 Well Plates)
  • 05:31CCF4 Assay on Live Samples Using Shigella flexneri (35 mm Glass-bottom Format, MatTek)
  • 07:04Results: The CCF4-AM/β-lactamase Approach to Pathogens
  • 09:48Conclusion

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.

Tags

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

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