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Immunology and Infection

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Nanyang Technological University

Microscopy-based Assays for High-throughput Screening of Host Factors Involved in Brucella Infection of Hela Cells

8.2K Views

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15:29 min

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August 5th, 2016

DOI :

10.3791/54263-v

August 5th, 2016

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Alain Casanova*¹, Shyan H. Low*¹, Mario Emmenlauer*¹^,⁴, Raquel Conde-Alvarez*¹^,³, Suzana P. Salcedo², Jean-Pierre Gorvel², Christoph Dehio¹

¹Focal Area Infection Biology, Biozentrum, University of Basel, ²Centre d’Immunologie de Marseille-Luminy, Université de la Méditérannée UM2, INSERM U1104 CNRS UM7280, ³Departmento de Microbiologìa and Instituto de Salud Tropical, Universidad de Navarra, ⁴BioDataAnalysis GmbH

Summary

Two assays for microscopy-based high-throughput screening of host factors involved in Brucella infection are described. The entry assay detects host factors required for Brucella entry and the endpoint assay those required for intracellular replication. While applicable for alternative approaches, siRNA screening in HeLa cells is used to illustrate the protocols.

Explore More Videos

High throughput Screening

Brucella Infection

Automated Workflow

Chapters in this video

0:05

Title

0:54

Reverse siRNA Transfection

3:19

Infection and Fixation

5:55

Staining and Imaging

8:14

Automated Image Analysis

12:21

Results: Automated High-throughput Quantification of Brucella Infection in HeLa Cells

14:06

Conclusion

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