改良的视网膜血管消化铺片联合共聚焦显微镜观察--视网膜血管的三维检查方法

目的探讨一种视网膜血管的三维检查方法。设计实验性研究。研究对象大鼠视网膜血管消化铺片。方法将大鼠深度麻醉后,行全身循环灌注,至眼球变苍白,再灌注4％多聚甲醛作血管内固定。取双眼以4％多聚甲醛固定24小时后,取出视网膜,改良视网膜血管消化铺片方法,采用胶原酶消化制备视网膜血管铺片,经免疫荧光染色后采用激光扫描共聚焦显微镜观察。主要指标视网膜血管铺片消化状况及三维结构的观察。结果胶原酶消化处理后可获得理想的、完整视网膜血管铺片, 通过激光扫描共聚焦显微镜扫描,可以观察到视网膜血管三维特征。结论改良的视网膜血管消化铺片联合共聚焦显微镜观察是一种有效的视网膜血管三维检查方法。

Modified retinal vascular digest preparations combined with laser scanning confocal microscope-a three-dimensional examination method for retinal vessels

Objective To explore a three-dimensional examination method for retinal vessels. Design Experimental study. Participants Retinal vascular digest preparations of rat. Methods After deep anaesthesia, rats were sacrificed and perfused till eyeballs pale. Then 4% paraformaldehyde was perfused for vascular internal fixation. Eyeballs were fixed in 4% paraformaldehyde for 24hr, and the retinal vessels were isolated with collagenase digest technique. After immunofluorescence staining, the retinal vessels were observed with laser scanning confocal microscope. Main Outcome Measures The digest state of retinal vascular digest preparations and three-dimensional observation of the preparations. Results Ideal complete retinal vascular digest preparations were obtained after collagenase digestion. Three-dimensional characteristics of retinal vessels can be observed with laser scanning confocal microscope. Conclusions Modified retinal vascular digest preparations combined with laser scanning confocal microscope provided us a method for retinal vascular three-dimensional structure observation.(phthalmol CHN,2006,15:138-141)