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Visualizing and Tracking T Cell Motility In Vivo.

Robert A Benson1, James M Brewer2, Paul Garside3

  • 1Centre for Immunobiology, Institute of Infection, Immunity and Inflammation, The University of Glasgow, Glasgow, G12 8TA, UK. robert.benson@glasgow.ac.uk.

Methods in Molecular Biology (Clifton, N.J.)
|March 29, 2017
PubMed
Summary

Advanced microscopy visualizes immune cell dynamics in lymph nodes. Researchers used multiphoton laser scanning microscopy for tracking CD4+ T cells and dendritic cells (DCs) in ex vivo and intravital settings.

Keywords:
Adoptive transferDendritic cellIn vivoIntravital imagingMigrationMultiphotonT cell

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Area of Science:

  • Immunology
  • Cell Biology
  • Microscopy

Background:

  • Immune responses involve complex cellular interactions within lymph nodes.
  • Understanding T cell and dendritic cell (DC) dynamics is crucial for immunology.
  • Advanced imaging techniques are needed to study these cellular behaviors in vivo.

Purpose of the Study:

  • To describe methods for visualizing and tracking fluorescently labeled CD4+ T cells and DCs in lymph nodes (LNs).
  • To compare ex vivo and intravital multiphoton laser scanning microscopy for studying immune cell responses.
  • To provide methods for quantitative, real-time analysis of cellular tracking data.

Main Methods:

  • Application of multiphoton laser scanning microscopy.
  • Ex vivo imaging of lymph nodes (LNs).
  • Intravital imaging of the popliteal LN.
  • Development of methods for time-series data acquisition for cellular tracking.

Main Results:

  • Successful visualization and tracking of CD4+ T cells and DCs within the LN environment.
  • Ex vivo imaging provides comparable data to intravital imaging without complex surgery.
  • Intravital imaging allows study within an intact lymph and blood supply context.
  • Acquired time-series data enables quantitative, real-time analysis of immune cell responses.

Conclusions:

  • Multiphoton microscopy is effective for studying immune cell dynamics in LNs.
  • Both ex vivo and intravital approaches offer valuable insights into T cell/DC interactions.
  • The described methods facilitate quantitative analysis of cellular tracking for immune response studies.