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A Multi-well Format Polyacrylamide-based Assay for Studying the Effect of Extracellular Matrix Stiffness on the Bacterial Infection of Adherent Cells
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
A Multi-well Format Polyacrylamide-based Assay for Studying the Effect of Extracellular Matrix Stiffness on the Bacterial Infection of Adherent Cells

A Multi-well Format Polyacrylamide-based Assay for Studying the Effect of Extracellular Matrix Stiffness on the Bacterial Infection of Adherent Cells

DOI:
10.3791/57361-v

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11:51 min

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July 05, 2018

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, , ,
1Department of Biochemistry,Stanford University School of Medicine, 2Department of Mechanical and Aerospace Engineering,University of California San Diego, 3Departments of Biochemistry, Microbiology and Immunology and Howard Hughes Medical Institute,Stanford University School of Medicine

Chapters

  • 00:05Title
  • 00:39Manufacturing Thin Two-layered Polyacrylamide Hydrogels on Multi-well Plates
  • 01:36Polyacrylamide Hydrogel Fabrication
  • 05:22Infection of Human Microvascular Endothelial Cells with L. monocytogenes
  • 07:11Flow Cytometry to Quantify Extracellular-matrix-stiffness Dependent Susceptibility of Host Cells to Infection
  • 08:02Quantitative Time-lapse Microscopy to Assess Extracellular-matrix-stiffness Dependent L. monocytogenes Dissemination Through Endothelial Cells
  • 09:01Results: The Effect of Extracellular Matrix Stiffness on L. monocytogenes Infection of HMEC-1 Cells
  • 10:32Conclusion

Summary

Automatic Translation

Automatic Translation

We have developed a multi-well format polyacrylamide-based assay for probing the effect of extracellular matrix stiffness on bacterial infection of adherent cells. This assay is compatible with flow cytometry, immunostaining, and traction force microscopy, allowing for quantitative measurements of the biomechanical interactions between cells, their extracellular matrix, and pathogenic bacteria.

Tags

Keywords: PolyacrylamideExtracellular MatrixStiffnessBacterial InfectionAdherent CellsHost-pathogen BiomechanicsHydrogelsAcrylamideBis-acrylamideTEMEDAPSFluorescent Microbeads

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