Abstract

FR.23.09

Real-time imaging of conjunctiva-associated lymphoid tissue (CALT) using in vivo two-photonmicroscopy

Steven P.
Institute of Anatomy, University of Lübeck and Eye Hospital, UK-SH, Campus Lübeck, Lübeck

Objective: Within the pathogenesis of Dry Eye Syndrom a relation to conjunctiva-associated lymphoid tissue (CALT) is discussed. Despite the fact that the morphology of CALT is well described, almost no functional immunological studies are available. Therefore descriptions of a functional relation between CALT and Dry Eye base on investigations of mucosa-associated lymphoid tissue (MATL) of other organs, e.g. the intestine. The presented project will demonstrate for the first time the feasibility of functional investigations of CALT by using an established mouse model and in vivo two-photon microscopy.
Methods: The Conjunctiva of female Balb/c mice is challenged with either Chlamydia trachomatis C or ovalbumin/choleratoxin B. The induced CALT will be investigated by in vivo two-photonmicroscopy.
Results: First results show that the application of either pathogen induces organized CALT at the nictating membrane of the eye. First real-time investigations using two-photon microscopy demonstrate autofluorescence images of all components of CALT (lymphoepithelium, follicles, blood and lymphatic vessels) up to 90um of depth without the use of artificial dyes and fixatives.
Conclusions: The established animal model enables functional investigations of CALT for the first time. Using real-time two-photon microscopy, tracking of lymphocytes and analyzing immunological processes within CALT allow future correlations of CALT function and Dry Eye Syndrom.

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