用于免疫荧光染色的大鼠视网膜冰冻切片的制作体会  

Production of Rat Retina Frozen Section Used for Immunofluorescence Staining

作　　者：郭岩[1] 孔佑华[1] 王凤琴[2] 

机构地区：[1]济宁医学院组胚教研室,山东济宁272067 [2]济宁医学院形态学实验室,山东济宁272067

出　　处：《沈阳医学院学报》2014年第3期162-163,172,共3页Journal of Shenyang Medical College

基　　金：2013年山东省高校青年骨干教师国内访问学者资助项目(No.鲁教人字(2013)8号)

摘　　要：目的：探索适合免疫荧光染色的大鼠视网膜冰冻切片的制作方法与技巧.方法：将SD大鼠麻醉后灌注或不灌注取眼球,根据标本处理差异分别进行固定及脱水处理,制作不同厚度视网膜冰冻切片60例,进行免疫荧光抗体标记,利用荧光显微镜观察其细胞排列及标记状况.结果：完整眼球视网膜结构形态可保持相对规则,但偶尔发生组织牵拉;玻璃体去除的视杯标本免疫荧光染色结果相对理想.标本厚度介于10～12 μm冰冻切片视网膜血管免疫荧光染色结果优于其他厚度者.结论：用于免疫荧光的组织标本应尽量保持其完整的固有形态,通过对目标结构的定性、定位以及定量,再现视网膜各成分的状态.实验者可根据需要选择最适合的眼组织取材方法、切片厚度等以便得到满意的观察效果.Objective： To explore optimal frozen section method and technique for immunofluorescence staining of rat retina. Methods： Eyes of SD rats were taken with perfusion or not after anesthesia, and fixed and dehydrated according to the various speci- men processing. A total of 60 frozen slices in different sickness were made, and then marked with immunohistochemical antibody. The cells arrangement and the status of the tag were observed with fluorescent microscope. Results： Full eye retina structure form could keep the relative rules, but organization pulling occasionally occurred. Immunofluorescence results of optic cup specimen after vitreous removal were relatively ideal. And the results of specimen thickness between 10 - 12 μm were superior to others. Conclu- sion ： Tissue samples for immunofluorescence should try to maintain its complete natural form by means of quantitative and qualitative, positioning of the target structure and reproduce style of the retina component.

关 键 词：SD大鼠 视网膜 冰冻切片 免疫荧光 

分 类 号：R774.13[医药卫生—眼科]