Structure of the erythrocyte membrane skeleton as observed by atomic force microscopy
Structure of the erythrocyte membrane skeleton as observed by atomic force microscopy
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DOI：
10.1016/s0006-3495(98)77926-3
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发表时间：
1998-05-01
影响因子：
3.4
通讯作者：
Kusumi, A
中科院分区：
文献类型：
--
作者：
Takeuchi, M;Miyamoto, H;Kusumi, A
利用原子力显微镜(AFM)对红细胞膜表面高度变化高度敏感的特点，用原子力显微镜(AFM)观察了干燥的人红细胞膜表面膜骨架的结构。膜骨架的大部分可以成像，甚至在膜的细胞外表面也可以成像。用不同的固定和干燥方法制备用于AFM观察的鬼膜样品，结果发现，对未固定的样品进行冷冻干燥(在冷冻剂中快速浸泡冷冻)是一种快速、简单的方法，可以在不去除膜或扩展膜骨架的情况下获得一致的良好观察结果。在细胞外表面观察膜骨架是可能的，主要是因为膜的双层部分在干燥过程中沉入细胞内。在细胞膜的胞外和胞浆表面观察到的平均网目尺寸分别为4800和3000 nm(2)，这表明在质膜的胞外表面可以观察到80%的网状结构，这表明在质膜的胞外表面可以观察到一个三维折叠的网络。
The structure of the membrane skeleton on the cytoplasmic surface of the erythrocyte plasma membrane was observed in dried human erythrocyte ghosts by atomic force microscopy (AFM), taking advantage of its high sensitivity to small height variations in surfaces. The majority of the membrane skeleton can be imaged, even on the extracellular surface of the membrane. Various fixation and drying methods were examined for preparation of ghost membrane samples for AFM observation, and it was found that freeze-drying (freezing by rapid immersion in a cryogen) of unfixed specimens was a fast and simple way to obtain consistently good results for observation without removing the membrane or extending the membrane skeleton. Observation of the membrane skeleton at the external surface of the cell was possible mainly because the bilayer portion of the membrane sank into the cell during the drying process. The average mesh size of the spectrin network observed at the extracellular and cytoplasmic surfaces of the plasma membrane was 4800 and 3000 nm(2), respectively, which indicates that spectrin forms a three-dimensionally folded meshwork, and that 80% of spectrin can be observed at the extracellular surface of the plasma membrane.