摘要
目的探讨一种简单且易于规范操作的小鼠血管灌注造影视网膜铺片技术。方法30只生后12～17d龄C57BL/6J小鼠麻醉后行20g.L-1伊文蓝上腔静脉灌注,5min后处死,摘取双眼眼球,40g.L-1多聚甲醛固定,A组20只眼球固定10min,B组20只眼球固定20min,C组20只眼球固定40min,手术显微镜下去除角膜、虹膜、晶状体、玻璃体及巩膜,并将视网膜平铺于载玻片上,荧光显微镜下照相,观察各组差异。结果大体观察:A组视网膜变大、变薄、变形,视网膜面积明显扩大;B组视网膜无变形,保持原有的形态;C组视网膜黏附黑色视网膜色素上皮层,不易分离。荧光显微镜下照相观察:A组铺片成功率为30%,视网膜血管变形,模糊不清;B组铺片成功率为90%,视网膜血管均匀规则分布;C组铺片成功率为30%,黑色视网膜色素上皮层部分黏附,血管背景成像不均匀。结论良好的规范操作技巧和恰当的多聚甲醛固定时间是保证小鼠血管灌注造影视网膜铺片成功的关键。
Objective To investigate an effective, simple and normative technique for stretched preparation of retina after angiography in mice. Methods Thirty C57BL/6J mice at postnatal 12 days to 17 days were anesthetized,and infused with 20 g· L^-1 Evans blue through the superior vena cava. After 5 minutes, the mice were killed, and the eyeballs of two eyes were enucleated. All eyeballs were divided randomly into three groups, 20 eyeballs in each group, and fixed in 40g· L^-1 paraformaldehyde for 10 minutes in group A,20 minutes in group B and 40 minutes in group C,respectively. After the cornea, iris, lens, vitreous body and sclera were removed under operating microscope ,the retina was tiled on the slide. The differences among three groups were observed with photograph under fluorescence microscope. Results Just with general observation, the retinas became big, thin and out of shape, and the area of retina became significantly large in group A. The shapes of retinas were unchanged and without disfiguration in group B. Black pigment epithelium layer adhered on the retinas, and not easily to be dissolved in group C. With photograph under fluorescence microscopy, the retina vessels were obscure and out of shape in group A;The retinal vessels had an uniform distribution in group B;Black pigment epithelium layer partly adhered, and the vascular background imaging was uneven in group C. The successful rate of stretched preparation was 30% in group A,90% in group B and 30% in group C, respectively. Conclusion A good, normative technique and correct time for fixation with paraformaldehyde are key factors for the success of stretched preparation of retina after angiography in mice.
出处
《眼科新进展》
CAS
北大核心
2009年第9期663-666,共4页
Recent Advances in Ophthalmology
基金
国家自然科学基金资助(编号:30772386)
科技部973计划前期研究专项基金资助(编号:2007CB516705)
山东省自主创新重大科技专项计划基金资助(编号:2006GG1102020)~~
关键词
血管造影
组织固定
视网膜铺片
小鼠眼球
angiography
tissue fixation
stretched preparation of retina
mouse eyeball
