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Related Experiment Video

Updated: Jan 14, 2026

Visualizing Lymph Node Structure and Cellular Localization using Ex-Vivo Confocal Microscopy
07:40

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Visualizing Endogenous Effector T Cell Egress from the Lymph Nodes.

Manisha Menon1, Alexandre P Benechet2, Kamal M Khanna3

  • 1Department of Immunology, University of Connecticut Health, 263 Farmington Ave, Farmington, CT, 06030, USA.

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

Studying T cell migration in lymphoid tissues during infection is crucial. This chapter details methods using intravital two-photon microscopy and in situ tetramer staining to visualize T cell dynamics in vivo.

Keywords:
In situ tetramer stainingIntravital microscopySphingosine-1-phosphate receptorT cell egress

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Area of Science:

  • Immunology
  • Cell Biology
  • Microscopy

Background:

  • Immune responses are regulated by the local anatomy of lymphoid tissues during infection.
  • Understanding cellular choreography within intact tissue environments is essential for studying immunity.
  • Pathogen-specific T cell migration and effector T cell egress from lymph nodes are complex, critical processes following infection.

Purpose of the Study:

  • To detail methods for visualizing the migration and egress of endogenous antigen-specific CD8 T cells in situ.
  • To investigate the mechanisms regulating T cell dynamics within lymphoid tissues during viral infection.
  • To provide a comprehensive assessment of cellular population distribution within intact tissues.

Main Methods:

  • Intravital two-photon (2P) microscopy for direct in vivo visualization of T cell dynamics.
  • Multicolor whole-mount in situ tetramer staining for static imaging and global cellular assessment.
  • Application of these techniques to study endogenous antigen-specific CD8 T cells following viral infection.

Main Results:

  • The study details methodologies for visualizing T cell migration and egress.
  • These methods allow for direct in vivo observation of T cell dynamics.
  • Static imaging provides a global assessment of cellular distribution within tissues.

Conclusions:

  • Intravital 2P microscopy and whole-mount in situ tetramer staining are powerful tools for studying T cell behavior in lymphoid tissues.
  • These techniques enable detailed investigation of cellular choreography during immune responses.
  • Understanding T cell dynamics in situ is crucial for advancing our knowledge of immunity and infection.