Abstract

SO.11.07

In-vivo imaging of lymphatic vessels in the cornea

Felix Bock1, Philipp Steven2, Gereon Hüttmann3, Claus Cursiefen1
1Universitäts-Augenklinik Erlangen, 2Universitäts-Augenklinik, Campus Lübeck, Universitätsklinikum Schleswig-Holstein, Lübeck, 3Institut für Biomedizinische Optik, Universität zu Lübeck, Lübeck

Objective
To visualize naturally invisible lymphatic vessels in the prevascularized, murine cornea in vivo with a multi-photon microscope.
Methods
FITC-conjugated LYVE-1 antibody was injected into a prevascularized murine cornea. Stained lymphatic vessels and additionally autofluorescent blood vessels, migrating cells and collagen fibrils were detected with a multi-photon microscope. The animals were anaesthetized and then placed in a special animal holder for controlled ventilation, temperature- and blood-oxygen monitoring.
Results
Lymphatic vessels could be visualized in the living animal and occurred as broad tubes with blind ends. 3D reconstruction of high resolution z-stacks demonstrated immune staining on the surface of the lymphatic endothelial cells accumulating at the cell-cell connections. Time series revealed cells migrating into the lymphatic vessels. Blood vessels (identified by fast moving erythrocyte signals), lymphatic vessels, migrating cells and collagen structures could be visualized simultaneously.
Conclusions
For the first time an intravital visualization of lymphatic vessels in the cornea was achieved. By this method we paved the way for new in vivo analyses of the interactions of the immune system and lymphatic vessels.

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