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A Rapid Image-based Bacterial Virulence Assay Using Amoeba
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JoVE Journal Bioingegneria
A Rapid Image-based Bacterial Virulence Assay Using Amoeba

A Rapid Image-based Bacterial Virulence Assay Using Amoeba

DOI:
10.3791/57844-v

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05:22 min

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June 27, 2018

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1Department of Physics and Astronomy,University of California, 2Department of Molecular Biology and Biochemistry,University of California

Capitoli

  • 00:04Titolo
  • 00:46Growth and Maintenance of Amoebae
  • 01:57The Growth of P. aeruginosa
  • 02:16Preparation of Agar Pad, Preparation of Bacteria-amoeba Sample, and Image Acquisition
  • 04:05Results: Virulence of P. aeruginosa is Upregulated in Surface-attached Populations
  • 04:38Conclusion

Summary

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Here, we present a protocol to measure the virulence of planktonic or surface-attached bacteria using D. discoideum (amoeba) as a host. Virulence is measured over a period of 1 h and host killing is quantified using fluorescence microscopy and image analysis. We demonstrate this protocol using the bacterium P. aeruginosa.

Tags

Bacterial VirulenceAmoebaQuantitative AssayHost-killingSurface-attached BacteriaPlanktonic CellsPseudomonas AeruginosaE. ColiAgar PadCalcium AMDevelopment Buffer

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