Related Experiment Video
Updated: Nov 4, 2025
Ex vivo Imaging of T Cells in Murine Lymph Node Slices with Widefield and Confocal Microscopes
Published on: July 15, 2011
Visualizing the Dynamics of T Cell-Dendritic Cell Interactions in Intact Lymph Nodes by Multiphoton Confocal
Billur Akkaya1, Olena Kamenyeva2, Juraj Kabat2
1Laboratory of Immune System Biology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA. billur.akkaya@nih.gov.
Insights
This study introduces a multiphoton microscopy protocol to visualize immune cell interactions within intact lymph nodes. The method allows simultaneous imaging of multiple cell types, including T cells and dendritic cells, in vivo.
Area of Science:
- Immunology
- Microscopy
- Cell Biology
Background:
- Multiphoton microscopy enables deep tissue visualization of cellular behavior in intact organs.
- Peripheral lymph nodes, such as the popliteal lymph node, are accessible for intravital imaging of immune cell dynamics.
- Understanding T cell-dendritic cell interactions is crucial for adaptive immunity.
Purpose of the Study:
- To describe a protocol for intravital imaging of antigen-specific T cell-dendritic cell interactions in the popliteal lymph node.
- To enable simultaneous visualization of multiple immune cell types with distinct specificities and lineages.
Main Methods:
- Utilizing multiphoton microscopy for deep tissue imaging.
- Performing intravital microscopy on the popliteal lymph node of immunocompetent mice.
- Developing a protocol to track antigen-specific T cells and dendritic cells simultaneously.
Main Results:
- The protocol allows for the visualization of T cell-dendritic cell interactions in vivo.
- Simultaneous imaging of up to four distinct cell types is achievable.
- This includes T cells with varying antigen specificities and differentiation states, as well as genetically modified dendritic cells.
Conclusions:
- The described protocol provides a powerful tool for studying immune cell behavior in lymph nodes.
- This method facilitates the in-depth analysis of T cell-dendritic cell interactions in their native microenvironment.
- It opens avenues for investigating immune responses in various physiological and pathological conditions.
Abstract:
Multiphoton microscopy has provided us the ability to visualize cell behavior and biology in intact organs due to its superiority in reaching deep into tissues. Because skin draining lymph nodes are readily accessible via minimal surgery, it is possible to characterize the intricate interactions taking place in peripheral lymph nodes intravitally. Here we describe our protocol to visualize antigen-specific T cell-dendritic cell interactions in the popliteal lymph node of immunocompetent mice. With this method, behaviors of up to four cell types, such as T cells with different antigen specificities, T cells differentiated into different effector and regulatory lineages and dendritic cells originating from mice that bear mutations in functional genes can be imaged simultaneously.
