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Updated: Dec 26, 2025
Intravital Imaging of the Mouse Popliteal Lymph Node
Published on: February 8, 2012
Multiphoton Intravital Microscopy of Mandibular Draining Lymph Nodes: A Mouse Model to Study Corneal Immune Responses
Maria J Lopez1,2, Yashar Seyed-Razavi1,2, Takefumi Yamaguchi1
1Department of Ophthalmology, Center for Translational Ocular Immunology, Tufts Medical Center, Tufts University School of Medicine, Boston, MA, United States.
Insights
This study introduces a new method for imaging immune cells in mandibular lymph nodes using multiphoton microscopy. This technique visualizes immune responses to corneal transplants, offering insights into ocular immunity.
Area of Science:
- Immunology
- Microscopy
- Ophthalmology
Background:
- Corneal immunity is vital for vision and transplant success.
- Understanding immune cell dynamics in draining lymph nodes is crucial.
- Previous intravital imaging of mandibular lymph nodes was challenging.
Purpose of the Study:
- To develop the first multiphoton intravital microscopy (MP-IVM) model for mouse mandibular lymph nodes (dLNs).
- To investigate immune cell behavior in dLNs following corneal transplantation.
- To explore the corneal immune reflex arc.
Main Methods:
- Utilized transgenic mice with fluorescently labeled CD11c+ cells.
- Applied MP-IVM to image mandibular dLNs in vivo.
- Analyzed cell density, speed, displacement, and directionality before and after corneal transplantation.
Main Results:
- Established a novel MP-IVM model for mandibular dLNs.
- Observed increased CD11c-YFP+ cell density and migration in dLNs post-corneal transplant.
- Demonstrated altered immune cell kinetics, including increased speed and directionality.
Conclusions:
- The developed MP-IVM technique enables visualization of immune cell spatiotemporal dynamics in mandibular dLNs.
- This model provides insights into the immune response to corneal transplantation.
- The technique is valuable for studying ocular immune and inflammatory diseases.
Abstract:
Multiphoton intravital microscopy (MP-IVM) is a powerful tool to image cells in vivo. Its application in immunology research has opened new horizons, allowing intravital imaging of leukocytes at the single-cell level. A transparent cornea is vital to retain vision. As an immune privileged site, a rapid innate response to foreign antigens is crucial in clearing opportunistic bacterial and viral pathogens, and minimizing collateral structural damage to the cornea. Furthermore, dissecting the mechanisms and preventing the immunological rejection process after corneal transplantation is imperative to retain sight. Therefore, understanding the underlying mechanisms behind corneal immunity, specifically the process of antigen presentation and adaptive immunity in the mandibular draining lymph nodes (dLNs) in vivo, is crucial. Attempts of intravital imaging of mandibular dLNs have yielded little success to date, due to breathing artifacts and the location that is difficult to access. Herein, we present the first MP-IVM mouse model of the mandibular dLNs, utilizing transgenic mice in which CD11c+ cells are fluorescently labeled. Furthermore, we demonstrate that CD11c-YFP+ cells are localized mainly in the parafollicular cortex (T cell zone) and subcapsular area and are sparsely distributed in the follicular region (B cell zone) of mandibular dLNs during steady state. A significant increase in host CD11c-YFP+ cell density is noted at 14 and 21 days following allogeneic corneal transplantation, compared to steady state (p < 0.05). Moreover, allogeneic corneal transplantation results in increased host-derived CD11c-YFP+ cell mean speed and displacement in mandibular dLNs, compared to steady state (p < 0.001). The meandering index, an index for directionality, is significantly increased after allogeneic corneal transplantation at both 14 and 21 days, compared to steady state (p < 0.001). Taken together, our study demonstrates the necessary methodology required for intravital multiphoton imaging of the mandibular dLNs, allowing visualization of spatiotemporal kinetics of immune cells in vivo, and provides a window into the corneal immune reflex arc. This technique will be a powerful tool to investigate the pathogenesis of ocular immune and inflammatory diseases.
