基于免疫荧光法对人工红细胞膜骨架蛋白spectrin的定位分析  

Immunofluorescence Technique for Location Analysis of Artificial Red Blood Cells Cytoskeletal Proteins-Spectrin

作　　者：王莹[1] 王翔[1] 李遥金[2] 刘峰[1] 肖科[1] 熊延连[1] 

机构地区：[1]教育部生物流变科学与技术重点实验室重庆大学生物工程学院,重庆400044 [2]第三军医大学西南医院,重庆400038

出　　处：《中国细胞生物学学报》2010年第3期429-432,共4页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(No.10572159);重庆市科委科技计划项目攻关项目(No.2006ba5010);高等学校学科创新引智计划(No.B06023);重庆大学大型仪器设备开放基金资助项目~~

摘　　要：本文建立免疫荧光方法对人工红细胞膜蛋白进行荧光定位,从膜结构蛋白角度分析本实验室构建的人工红细胞,也为血液代用品的评价提供了新的途径。以荧光标记兔抗人免疫球蛋白为抗体,采用间接免疫荧光染色法对红细胞膜蛋白spectrinα1原位标记,在激光共聚焦显微镜下以488 nm蓝光激发荧光,通过对比天然红细胞和人工红细胞的spectrin分布,分析人工红细胞的构象和重建的过程。与天然红细胞类似,人工红细胞在膜上有血影蛋白分布,本实验荧光原位标记结果进一步揭示了人工红细胞的体外构建过程。In this paper, we reported a new way to locate membrane protein of artificial red blood cells （RBC） and also provided a new method to estimate blood substitute by using immunofluorescence technique. By appling this method, we analyzed the artificial RBC reconstructed by our laboratory from the view of membrane structural proteins. The conformation of artificial RBC and its reconstrucing processes were analyzed by the comparison of distribution of spectrin in both natural RBC and artificial RBC, while spectrin α1 was coloured in situ by indirect immunofluorescence method using fluorescence labeled rabbit-anti-human immunoglobulina and the fluorescence detected with laser scanning confocal microscope （LSCM） was excited by blue light of 488nm wavelength. It is similar to natural RBC, spectrin also distributed in the cytomembrane of RBC. Besides, our results further revealed the reconstructing processes of artificial RBC in vitro.

关 键 词：免疫荧光法 人工红细胞 血影蛋白 定位 

分 类 号：Q26[生物学—细胞生物学]