小鼠视网膜组织石蜡切片制作方法的探讨  

Investigation of fixative method of mouse retinal paraffin section

作　　者：程钧[1] 万磊[1] 刘廷[1] 刘伟吉[1] 董晓光[1] 

机构地区：[1]山东省眼科研究所,山东省青岛市266071

出　　处：《眼科新进展》2009年第3期161-164,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)~~

摘　　要：目的寻找一种效果好的视网膜固定方法。方法30只小鼠取出眼球后在角巩膜缘刺一小口,A组24只眼球,用体积分数10%中性甲醛固定;B组16只眼球,用FAA固定液(体积分数10%中性甲醛10mL、体积分数95%乙醇85mL、冰醋酸5mL混合)固定;C组20只眼球,置于FAA固定液中浸泡1min后用体积分数10%中性甲醛固定过夜,脱水后去除角膜、虹膜和晶状体,分别做HE染色和免疫组织化学染色,镜下观察其差异。结果大体观察A组大部分眼球皱缩变形,视网膜脱离率高;B组、C组眼球形态均无明显变化,无视网膜脱离发生。HE染色示A组标本视网膜结构疏松,有较多裂隙,且易发生视网膜碎裂;B组标本视网膜过度压缩,视网膜各层结构不清晰;C组标本视网膜结构清晰,细胞排列紧密,色泽艳丽。免疫组织化学染色3组均显示神经元特异性烯醇化酶NSE阳性,但C组标本染色效果优于A组、B组,且脱片程度低,适用于视网膜组织的固定。结论采用FAA先浸泡再用体积分数10%中性甲醛固定视网膜组织的效果优于单独使用体积分数10%中性甲醛和FAA固定液。Objective To search for an effective fixative method for mouse retina.Methods Eyeballs of 30 mice were taken out,and pricked for a small hole at the corneoscleral limbus.All eyeballs were divided randomly into 3 groups,twenty-four eyeballs in group A fixed in 10% neutral formalin,sixteen eyeballs in group B fixed in an admixture of 10% neutral formalin（10 mL）,95% alcohol（85 mL） and glacial acetic acid （FAA,5 mL）,twenty eyeballs in group C soaked into FAA for 1 minute and then fixed in 10% neutral formalin and stayed overnight.Then the eyeballs were anhydration and removed cornea,iris,and lens.HE staining and immunohistochemistry were performed,and then compared the differences under the light microscope.Results In general,a great many of eyeballs in group A were shrunk and the rate of retinal detechment was very high;The shape of eyeballs in group B and C were well and no retinal detachment happened.HE staining showed that the retinal structures of group A were raritas and there were many fragmentation.The retinas of group B contracted too much so that the structures were not clear.In group C,the structures of the retinas were clear,the cells were arranged and compacted,and the colour was beautiful.Immunohistochemistry showed positive expression of NSE in 3 groups,but group C was with a better result than that of group A or B,and few exfoliation of the tissue from the glass slide was observed,which was good for retinal tissue fixation.Conclusion The method,soaked into FAA first and then fixed in 10% neutral formalin,is superior to using 10% neutral formalin or FAA fixation alone.

关 键 词：视网膜 组织固定 石蜡切片 小鼠眼球 

分 类 号：R774.1[医药卫生—眼科]