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High Throughput, Real-time, Dual-readout Testing of Intracellular Antimicrobial Activity and Eukaryotic Cell Cytotoxicity
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Immunology and Infection
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JoVE Journal Immunology and Infection
High Throughput, Real-time, Dual-readout Testing of Intracellular Antimicrobial Activity and Eukaryotic Cell Cytotoxicity

High Throughput, Real-time, Dual-readout Testing of Intracellular Antimicrobial Activity and Eukaryotic Cell Cytotoxicity

DOI:
10.3791/54841-v

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09:09 min

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November 16, 2016

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, ,
1Department of Pathology,Beth Israel Deaconess Medical Center

Chapters

  • 00:05Title
  • 01:30Real Time Intracellular Growth and Eukaryotic Cell Cytotoxicity Assay
  • 03:14Preparing Luminescent or Fluorescent Legionella pneumophila and Macrophage Infection
  • 04:49Assay Readout, and Single and Multi-dimensional Dose-response Testing and Data Interpretation
  • 06:09Results: Simultaneous Real-time Assay Results for Intracellular Bacterial Growth and Eukaryotic Cell Cytotoxicity
  • 07:41Conclusion

Summary

Automatic Translation

Automatic Translation

A high throughput, real-time assay was developed to simultaneously identify (1) eukaryotic cell-penetrant antimicrobials targeting an intracellular bacterial pathogen, and (2) assess eukaryotic cell cytotoxicity. A variation on the same technology was thereafter combined with digital dispensing technology to enable facile, high-resolution, dose-response, and two- and three-dimensional synergy studies.

Tags

Keywords: Intracellular Antimicrobial ActivityEukaryotic Cell CytotoxicityReal-time AssayHigh-throughputDual-readoutAntimicrobial DiscoveryMacrophageJ774A.1 CellsRPMI 1640 MediumLiquid Handling384-well Microplates

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