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CLIQ-BID: A method to quantify bacteria-induced damage to eukaryotic cells by automated live-imaging of bright nuclei

Yann Wallez, Stéphanie Bouillot, Emmanuelle Soleilhac, Philippe Huber, Ina Attrée, Eric Faudry
doi: https://doi.org/10.1101/236091
Yann Wallez
1UGA, CEA, INSERM, CNRS, Bacterial Pathogenesis and Cellular Responses, Biosciences and Biotechnology Institute of Grenoble, France.
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Stéphanie Bouillot
1UGA, CEA, INSERM, CNRS, Bacterial Pathogenesis and Cellular Responses, Biosciences and Biotechnology Institute of Grenoble, France.
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Emmanuelle Soleilhac
2UGA, CEA, INSERM; Genetics & Chemogenomics, CMBA Platform, Biosciences and Biotechnology Institute of Grenoble, France
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Philippe Huber
1UGA, CEA, INSERM, CNRS, Bacterial Pathogenesis and Cellular Responses, Biosciences and Biotechnology Institute of Grenoble, France.
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Ina Attrée
1UGA, CEA, INSERM, CNRS, Bacterial Pathogenesis and Cellular Responses, Biosciences and Biotechnology Institute of Grenoble, France.
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Eric Faudry
1UGA, CEA, INSERM, CNRS, Bacterial Pathogenesis and Cellular Responses, Biosciences and Biotechnology Institute of Grenoble, France.
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  • For correspondence: eric.faudry@cea.fr
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ABSTRACT

Pathogenic bacteria induce eukaryotic cell damage which range from discrete modifications of signalling pathways, to morphological alterations and even to cell death. Accurate quantitative detection of these events is necessary for studying host-pathogen interactions and for developing strategies to protect host organisms from bacterial infections. Investigation of morphological changes is cumbersome and not adapted to high-throughput and kinetics measurements. Here, we describe a simple and cost-effective method based on automated analysis of live cells with stained nuclei, which allows real-time quantification of bacteria-induced eukaryotic cell damage at single-cell resolution. We demonstrate that this automated high-throughput microscopy approach permits screening of libraries composed of interference-RNA, bacterial strains, antibodies and chemical compounds in ex vivo infection settings. The use of fluorescently-labelled bacteria enables the concomitant detection of changes in bacterial growth. Using this method named CLIQ-BID (Cell Live Imaging Quantification of Bacteria Induced Damage), we were able to distinguish the virulence profiles of different pathogenic bacterial species and clinical strains.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted December 18, 2017.
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CLIQ-BID: A method to quantify bacteria-induced damage to eukaryotic cells by automated live-imaging of bright nuclei
Yann Wallez, Stéphanie Bouillot, Emmanuelle Soleilhac, Philippe Huber, Ina Attrée, Eric Faudry
bioRxiv 236091; doi: https://doi.org/10.1101/236091
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CLIQ-BID: A method to quantify bacteria-induced damage to eukaryotic cells by automated live-imaging of bright nuclei
Yann Wallez, Stéphanie Bouillot, Emmanuelle Soleilhac, Philippe Huber, Ina Attrée, Eric Faudry
bioRxiv 236091; doi: https://doi.org/10.1101/236091

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