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Updated: May 29, 2026

Live-cell Imaging of Migrating Cells Expressing Fluorescently-tagged Proteins in a Three-dimensional Matrix
10:26

Live-cell Imaging of Migrating Cells Expressing Fluorescently-tagged Proteins in a Three-dimensional Matrix

Published on: December 22, 2011

High-resolution fluorescence microscopy to study transendothelial migration.

Christopher V Carman1

  • 1Center for Vascular Biology Research, Division of Molecular and Vascular Medicine, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA. ccarman@bidmc.harvard.edu

Methods in Molecular Biology (Clifton, N.J.)
|September 13, 2011
PubMed
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Understanding how immune cells, or leukocytes, cross the endothelium (blood vessel lining) is key to immunity. This study provides protocols for in vitro models to visualize leukocyte transendothelial migration (TEM) using advanced imaging techniques.

Area of Science:

  • Immunology
  • Cell Biology
  • Biophysics

Background:

  • Immune cell trafficking, specifically leukocyte transendothelial migration (TEM), is crucial for immune surveillance and pathogen response.
  • The endothelium forms a selective barrier between blood circulation and tissues, regulating immune cell movement.
  • Understanding the mechanisms of TEM is fundamental to comprehending immune system function.

Purpose of the Study:

  • To provide protocols for establishing in vitro models to study leukocyte transendothelial migration (TEM).
  • To detail methods for high-resolution live-cell and 3D fixed-cell imaging of TEM.
  • To facilitate the study of dynamic cellular behaviors during leukocyte migration across the endothelium.

Main Methods:

  • Utilizing cultured endothelial cell monolayers and isolated blood leukocytes to create in vitro TEM models.

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

Published on: August 14, 2014

Related Experiment Videos

Last Updated: May 29, 2026

Live-cell Imaging of Migrating Cells Expressing Fluorescently-tagged Proteins in a Three-dimensional Matrix
10:26

Live-cell Imaging of Migrating Cells Expressing Fluorescently-tagged Proteins in a Three-dimensional Matrix

Published on: December 22, 2011

Real-time Imaging of Endothelial Cell-cell Junctions During Neutrophil Transmigration Under Physiological Flow
11:26

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

Published on: August 14, 2014

  • Employing high spatial and temporal resolution imaging techniques for dynamic cell observation.
  • Implementing protocols for both live-cell imaging and three-dimensional fixed-cell imaging of TEM.

    • Main Results:

    • Established a basic in vitro system for studying transendothelial migration (TEM).
    • Demonstrated the utility of advanced imaging for capturing dynamic leukocyte and endothelial cell behaviors during TEM.
    • Revealed complex cellular dynamics influencing TEM efficiency.

    Conclusions:

    • In vitro models combined with high-resolution imaging offer powerful tools to investigate leukocyte transendothelial migration (TEM).
    • These methods enable detailed analysis of the dynamic cellular interactions governing immune cell movement across the endothelium.
    • The provided protocols support further research into immune system function and regulation.