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Don Yuen, Xiufeng Wu, Tatiana Ecoiffier, Alex Kwan, Lu Chen; Live Imaging of Lymphatic Vessels in the Cornea. Invest. Ophthalmol. Vis. Sci. 2012;53(14):2393.
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Lymphatic system plays an important role in tissue homeostasis and immune responses. Its dysfunction has been found in many disorders from cancer metastasis to transplant rejection. The cornea provides an ideal site for lymphatic research due to its accessible location, transparent nature, and lymphatic-free but -inducible features. An advanced technology for live imaging of lymphatic vessels in this tissue would therefore have widespread applications in biomedical research. In this study, we present a novel and highly efficient method for live imaging of newly formed lymphatic vessels in murine cornea, one of the most desirable sites for vascular research.
Corneal lymphangiogenesis was induced by suture placement or micropocket implantation. Newly formed lymphatic vessels were visualized by large molecule FITC-dextran injection. Dye uptake was monitored under a custom-built live imaging system with an adjustable eye and head holder to secure steady pictures. The specificity of FITC-dextran labeling of lymphatic vessels in the cornea was examined by immunofluorescent staining with LYVE-1, the lymphatic marker.
The new live imaging technique generated high quality pictures showing lymphatic vessels in clear contrast against the background within live mouse cornea. It produced multi-dimensional images of lymphatic vessels in corneal stroma. The system can be used to demonstrate lymphatic vessels at various stages and to capture lymphatic changes in the same cornea over a certain time period. The FITC-dextran labeled vessels expressed LYVE-1, confirming their lymphatic nature.
The new live imaging system provides a powerful tool to study dynamic lymphatic processes in the cornea. It possesses major advantages over conventional immunohistochemistry method, which is non-applicable for time course evaluation of the same tissue. Given its high efficiency and accuracy, we anticipate this new technique will facilitate a number of applications in biomedical research, such as dissection of lymphatic processes from formation to regression, or examination of the therapeutic effect of a pharmaceutical product.
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