两种组织透明技术在免疫荧光染色观察脊髓3D结构中的应用  

Application of Two Kinds of Tissue Clearing Method in Observing 3D Imaging of Spinal Cord with Immunofluorescent Staining

作　　者：段红梅[1] 尚俊奎 郝鹏[2] 李千千[1] 杨飞祥[2] 杨朝阳[1,2] 李晓光[1,2] 

机构地区：[1]北京航空航天大学生物与医学工程学院,北京市100191 [2]首都医科大学神经生物学系,北京市100069

出　　处：《中国康复理论与实践》2016年第4期417-421,共5页Chinese Journal of Rehabilitation Theory and Practice

基　　金：国家"863"计划项目(No.2012AA020506);国家自然科学基金面上项目(No.31271037);国家自然科学基金国际合作与交流项目(No.31320103903);国家自然科学基金委员会资助项目(No.31130022);"十二五"国家科技支撑计划项目(No.2012BAI17B04);高等学校全国优秀博士学位论文作者专项资金资助项目(No.201356);国家国际科技合作专项项目(No.2014DFA30640)

摘　　要：目的观察CUBIC和i DISCO两种透明技术在脊髓免疫荧光染色中的效果。方法应用两种组织透明技术处理1 mm厚脊髓冠状切片,然后行神经丝(NF)蛋白免疫荧光染色,激光共聚焦扫描显微镜观察染色效果。结果与CUBIC透明相比,i DISCO透明技术所需时间短,透明度高(F=6.64,P<0.01),免疫荧光染色深(F=5117.55,P<0.01)。结论应用i DISCO透明方法和免疫荧光染色相结合的方法可以完整观察脊髓轴突,更加简单快速,染色效果更好。Objective To compare the application of CUBIC and i DISCO clearing methods in observing 3D imaging of spinal cord with immunofluorescent staining. Methods 1 mm thick spinal cord coronal sections were processed with CUBIC and i DISCO, respectively. The neurofilament（NF） protein was detected by immunofluorescent staining and then was observed by a laser confocal microscope. Results Compared with CUBIC, i DISCO had the advantages of shorter time, higher transparency（F=6.64, P〈0.01）, and deeper penetration（F=5117.55, P〈0.01）. Conclusion Immunofluorescent staining combined with i DISCO could completely observe the spinal axons with shorter time and better stain effect.

关 键 词：脊髓 透明 轴突再生 CUBIC iDISCO 大鼠 

分 类 号：R651.2[医药卫生—外科学]