中文摘要：

目的探讨标记大鼠全角膜新生血管和淋巴管的方法。方法应用心脏灌注荧光联合全角膜免疫荧光法．共焦显微镜下区分并标记全角膜的新生血管和淋巴管。结果适度的抗体浓度和灌注时间下可标记出全角膜新生血管和淋巴管。血管呈黄绿色，淋巴管呈红色。结论心脏灌注荧光联合全角膜免疫荧光法可用于标记大鼠全角膜新生血管和淋巴管。

英文摘要：

Objective Corneal lymphangiogenesis provides an exit route for antigen presenting cells to the regional lymph node inducing immune response. However, corneal lymphangiogenesis is difficult to identify and visualize clinieally The purpose of this study was to explore a novel method to visualize corneal new lymphatic vessels in rat whole mount corneas. Methods Corneal alkali burned model was made in both eyes of 26 rats by sticking filter soaked in 1 mol/L NaOH solution to cornea,and 3 normal rats was as controls. Corneal new blood vessels and lymphatic vessels were identified by electron microscopyl ,2,3 days postinjury. Gradient concentration of platelet-endothelial cell adhension molecule/phycoerythrin （PECAM-PE） （ 1 ： 500 - 1 ： 25 ） was used to visualize corneal vessels. Whole cornea was incubated with PE-conjugated anti-PECAM （ 1： 50）and Lectin-FITC was injected through carotid artery for 30 seconds and 1 ,2,3,4 minutes respectively to perform cofocal microscopic examination. Results Both corneal hemangiogenesis and lymphangiogenesis were seen in stroma 3 days after initiation of model under the electron microscopy and clear red fluorescence for 1 ： 50 PE-conjugated anti-PECAM could be seen. The results of FITCconjugated lectin labeling showed that new blood vessels appeared in yellow-green fluorescence 2 minutes after profusion and the new lymphatic vessels has red fluorescence. Conclusion Combination of heart profusion and corneal immunofluorescence is a good approach to identify new blood vessels and lymphatic vessels on whole mount cornea of rat.