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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
DOI:

10:39 min

June 12, 2013

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Chapters

  • 00:05Title
  • 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

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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