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Immunology and Infection
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Real-time Imaging of Endothelial Cell-cell Junctions During Neutrophil Transmigration Under Physiological Flow
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Real-time Imaging of Endothelial Cell-cell Junctions During Neutrophil Transmigration Under Physiological Flow

Real-time Imaging of Endothelial Cell-cell Junctions During Neutrophil Transmigration Under Physiological Flow

DOI:
10.3791/51766-v

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11:26 min

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August 14, 2014

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, , ,
1Department of Molecular Cell Biology,Sanquin Research and Landsteiner Laboratory, AMC at University of Amsterdam

Chapters

  • 00:05Title
  • 01:59Plating and Maintenance of Human Umbilical Vein Endothelial Cells
  • 04:00Polymorphonuclear Leukocyte Isolation Using Percoll Gradients
  • 06:36Labeling of Endothelial Junctional VE-cadherin and PECAM-1 and Performing the PMN TEM Assay Under Flow
  • 09:24Results: Visualization of the Dynamics of VE-cadherin and PECAM-1 During Neutrophil TEM
  • 10:59Conclusion

Summary

Automatic Translation

Automatic Translation

Leukocytes cross the endothelial monolayer using the paracellular or the transcellular route. We developed a simple assay to follow the distribution of endogenous junctional VE-cadherin and PECAM-1 during leukocyte transendothelial migration under physiological flow to discriminate between the two transmigration routes.

Tags

Keywords: Endothelial Cell-cell JunctionsNeutrophil TransmigrationPhysiological FlowLeukocyte Transendothelial MigrationParacellular RouteTranscellular RouteVE-cadherinPECAM-1In Vitro AssayFluorescent AntibodiesElectrical Cell-substrate Impedance SensingFRAP Measurements

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