视网膜血管铺片技术的改进

摘要

Objective To improve the method of retinal vascular preparation and application in experimental animals of mice, rats, dogs and monkeys.Methods The retinal vessels were isolated with proteinase K-trypsin digestion technique.The samples were stained by hematoxylin-eosin(HE) and Periodic-Acid Schiff(PAS).The expression of CD31 and VEGF were detected by immunohistochemistry(IHC).Results The spread sheets of vascular network are complete, and can clearly show the blood vessels.No nerve tissue residue is left.Different levels of vascular morphology are clear. The vascular branches are clear and complete with no fracture.The cell morphology is intact, and the cell nucleus is clearly dispayed.The entire process can be completed in a short time, and has a high success rate.The vascular antigen are successfully retained: Expression of CD31 can be seen in both pericytes and endothelial cells; VEGF is mainly expressed in endothelial cells.Conclusions The improved method is successfully applied in different experimental animals.This approach will provide an important contribution to the retinal vascular disease research.%目的：探讨视网膜血管铺片技术的改进及在实验动物小鼠，大鼠，狗和猴中的应用。方法应用蛋白酶 K －胰蛋白酶联合消化法制备视网膜血管铺片，行 HE 及 PAS 染色观察血管形态，免疫组化方法观察 CD31及VEGF 的表达。结果用改进方法制备的视网膜血管铺片，血管网完整，无神经组织残留，可清晰显示血管走向。各级血管形态清晰，血管分支清楚完整，无断裂现象。血管内皮细胞形态清晰。 CD31在血管周细胞及内皮细胞均有表达；VEGF 主要在内皮细胞表达。结论改进的视网膜血管铺片技术可成功应用于不同实验动物，为视网膜血管性疾病的研究提供了重要途径。