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Updated: Nov 16, 2025
Ex vivo Imaging of T Cells in Murine Lymph Node Slices with Widefield and Confocal Microscopes
Published on: July 15, 2011
Acute Lymph Node Slices Are a Functional Model System to Study Immunity Ex Vivo
Maura C Belanger1,2, Alexander G Ball3,2, Megan A Catterton1
1Department of Chemistry, University of Virginia College of Arts and Sciences, Charlottesville, Virginia 22904, United States.
Insights
Live lymph node slices preserve tissue complexity for studying adaptive immunity. This validated platform maintains cell viability and function, enabling detailed immune response analysis.
Area of Science:
- Immunology
- Cell Biology
- Tissue Engineering
Background:
- Lymph nodes are crucial for adaptive immunity, but ex vivo studies often lose spatial organization or dynamic accessibility.
- Existing methods like cell suspensions or fixed tissues limit comprehensive analysis of lymph node functions.
- Live murine lymph node slices offer a potential solution to retain tissue complexity and enable dynamic studies.
Purpose of the Study:
- To systematically characterize live murine lymph node slices as a validated platform for studying immune responses.
- To assess the viability, immune activation, and antigen-specific functions of these ex vivo tissue slices.
- To establish optimized conditions for preparing and utilizing lymph node slices in immunological research.
Main Methods:
- Systematic characterization of live murine lymph node slices.
- Assessment of spatial organization and cell populations.
- Evaluation of 24-h viability, inflammation markers, and immune cell activation.
- Functional assays including T cell receptor cross-linking and antigen-specific cytokine secretion.
- Comparison with traditional cell suspension methods.
Main Results:
- Live lymph node slices successfully maintained spatial organization and native cell populations.
- Optimized slices demonstrated comparable 24-h viability and inflammation levels to cell suspensions.
- Slices exhibited robust responses to T cell receptor stimulation, including increased surface marker expression and cytokine secretion.
- The tissue slices processed protein antigens and showed antigen-specific cytokine secretion in response to ex vivo challenge in vaccinated animals.
Conclusions:
- Live murine lymph node slices represent a versatile and validated platform for immunological research.
- This method preserves the spatial complexity and dynamic accessibility of lymph nodes for ex vivo studies.
- Lymph node slices enable well-defined stimulation, time-course analysis, and parallel read-outs of immune functions.
Abstract:
The lymph node is a highly organized and dynamic structure that is critical for facilitating the intercellular interactions that constitute adaptive immunity. Most ex vivo studies of the lymph node begin by reducing it to a cell suspension, thus losing the spatial organization, or fixing it, thus losing the ability to make repeated measurements. Live murine lymph node tissue slices offer the potential to retain spatial complexity and dynamic accessibility, but their viability, level of immune activation, and retention of antigen-specific functions have not been validated. Here we systematically characterized live murine lymph node slices as a platform to study immunity. Live lymph node slices maintained the expected spatial organization and cell populations while reflecting the 3D spatial complexity of the organ. Slices collected under optimized conditions were comparable to cell suspensions in terms of both 24-h viability and inflammation. Slices responded to T cell receptor cross-linking with increased surface marker expression and cytokine secretion, in some cases more strongly than matched lymphocyte cultures. Furthermore, slices processed protein antigens, and slices from vaccinated animals responded to ex vivo challenge with antigen-specific cytokine secretion. In summary, lymph node slices provide a versatile platform to investigate immune functions in spatially organized tissue, enabling well-defined stimulation, time-course analysis, and parallel read-outs.
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