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Updated: Aug 8, 2026
Long-term Intravital Immunofluorescence Imaging of Tissue Matrix Components with Epifluorescence and Two-photon Microscopy
Published on: April 22, 2014
Imaging ocular immune responses by intravital microscopy
James T Rosenbaum1, Stephen R Planck, Tammy M Martin
1Casey Eye Institute, Department of Medicine-Division of Allergy, Immunology and Rheumatic Diseases, Department of Cell & Developmental Biology, Oregon Health and Science Center, Portland, Oregon, USA.
Insights
The eye allows real-time observation of immune cell interactions, including leukocyte trafficking and adhesion, using advanced microscopy. These insights into ocular immune responses are crucial for understanding eye diseases and evaluating treatments.
Area of Science:
- Ophthalmology
- Immunology
- Microscopy
Background:
- The eye provides a unique in vivo environment for studying cellular interactions.
- Immune responses within the eye involve complex events like cell trafficking, adhesion, and antigen presentation.
Purpose of the Study:
- To investigate immune cell behavior and interactions within the eye using intravital microscopy.
- To analyze leukocyte-endothelial interactions in ocular microvascular beds.
- To determine the timing of T cell trafficking and identify antigen-presenting cells in ocular tissues.
Main Methods:
- Intravital microscopy of the conjunctiva, cornea, and iris.
- Scanning laser ophthalmoscopy in rat and mouse models.
- Real-time imaging of fluorescently labeled antigens and cells in the anterior chamber.
- Analysis of leukocyte-endothelial interactions in retinal and choroidal circulations.
Main Results:
- Detailed observation of leukocyte rolling and adhesion in unmanipulated ocular microvasculature.
- Quantification of shear stress effects on leukocyte-endothelial adhesion.
- Insights into T cell trafficking dynamics and potential antigen-presenting cells within the eye.
Conclusions:
- Intravital microscopy offers powerful tools for studying ocular immune responses in real time.
- These methods provide valuable data on leukocyte behavior and antigen presentation in the eye.
- The techniques are being adapted for human eye studies to understand tissue damage and treatment efficacy.
Abstract:
The eye offers excellent opportunities to observe cellular interactions in vivo. This applies especially to the immune response in which discrete events can be studied, including cell trafficking, transendothelial migration, adhesion, antigen presentation, and T cell activation. Intravital microscopy has allowed study of immune cell interactions in tissues such as the conjunctiva, the inflamed cornea, and the iris. Thus the realtime observation of presentation of antigen injected into the anterior chamber of the eye can be imaged using fluorescently labelled antigen and cells. Application of the scanning laser ophthalmoscope to the rat and mouse eye allows analysis of leukocyte-endothelial interactions in the retinal and choroidal circulations. These studies have provided important information on rolling and adhesion of leukocytes in real time in different microvascular beds that have not been manipulated in any way and has, for instance, provided quantitation to the effects of shear stress on leukocyte-endothelial adhesion. In addition, the model permits an accurate analysis of the timing of trafficking of T cells into the eye and the possibility of determining which cells, if any, may be responsible for antigen presentation in the tissues as opposed to the secondary lymphoid organs. Finally, these experimental methods are now being applied to the human eye and should prove valuable in determining the nature of tissue damage events in the eye as well as evaluating the response to treatments.
