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A Microscopic Phenotypic Assay for the Quantification of Intracellular Mycobacteria Adapted for High-throughput/High-content Screening
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Immunology and Infection
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JoVE Journal Immunology and Infection
A Microscopic Phenotypic Assay for the Quantification of Intracellular Mycobacteria Adapted for High-throughput/High-content Screening

A Microscopic Phenotypic Assay for the Quantification of Intracellular Mycobacteria Adapted for High-throughput/High-content Screening

DOI:
10.3791/51114-v

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15:28 min

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January 17, 2014

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, , , , , , , ,
1Inserm U1019 – CNRS UMR 8024, Institut Pasteur de Lille,Université de Lille

Chapters

  • 00:05Title
  • 01:44Preparation of the Bacterial Suspension
  • 03:07High-throughput Genome-wide siRNA Screening
  • 06:16High-throughput Compound Screening
  • 08:27Image Acquisition and Analysis
  • 11:10Results: Mtb Colonization in Host Cells is Inhibited by Silencing Dectin-1 and by Isonazid or Rifampicin Treatment
  • 14:37Conclusion

Summary

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Here, we describe a phenotypic assay applicable to the High-throughput/High-content screens of small-interfering synthetic RNA (siRNA), chemical compound, and Mycobacterium tuberculosis mutant libraries. This method relies on the detection of fluorescently labeled Mycobacterium tuberculosis within fluorescently labeled host cell using automated confocal microscopy.

Tags

Keywords: TuberculosisMycobacteriaIntracellularPhenotypic AssayHigh-throughput ScreeningHigh-content ScreeningMycobacterium TuberculosisDrug ResistanceGenetic ScreensChemical ScreensFluorescence MicroscopyImage AnalysisHost-pathogen Interaction

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