用金溶胶技术显示晶状体上皮细胞的细胞骨架

Demonstration of the cytoskeleton of lens epithelial cells with gold sol techniques.

Spindler M, Iwig M

Institute of Biochemistry, Martin-Luther-University, Halle/Saale, Germany.

Acta Histochem Suppl. 1991;41:153-9.

Abstract

The Triton-extraction procedure is well suited for the demonstration of the cytoskeleton by TEM. A good preservation of the fine cytoskeletal network depends on optimal drying. 2. Immunogold-labeling is a useful method for visualization of intermediate filaments (IF) and microfilaments (MF). Labelled and unlabelled filaments are discernible. 3. Various methods were tested for a direct gold-labeling of MF with phallotoxin-gold preparations: The application of phalloidin-gold complexes results in an intensive unspecific staining of cytoskeletal filaments. A modified coupling procedure for the binding of phallacidin to transferrin was necessary to avoid the agglomeration of gold particles. The direct labeling of the cytoskeleton by the application of phallacidin-transferrin-gold complexes to Triton-extracted cells demonstrated an enrichment of gold particles on the both types of filaments, but MF and IF were not discernible. The treatment with phallacidin-polylysin-gold complexes demonstrated labelled filaments, but without sufficient intensity.

摘要

曲拉通提取法非常适合通过透射电子显微镜（TEM）来展示细胞骨架。精细细胞骨架网络的良好保存取决于最佳干燥条件。2. 免疫金标记是一种用于可视化中间丝（IF）和微丝（MF）的有用方法。标记和未标记的丝是可辨别的。3. 测试了多种用鬼笔环肽-金制剂直接对微丝进行金标记的方法：应用鬼笔环肽-金复合物会导致细胞骨架丝出现强烈的非特异性染色。为避免金颗粒的聚集，需要一种用于将鬼笔环肽与转铁蛋白结合的改良偶联程序。将鬼笔环肽-转铁蛋白-金复合物应用于经曲拉通提取的细胞对细胞骨架进行直接标记，结果表明在这两种类型的丝上都有金颗粒富集，但微丝和中间丝无法辨别。用鬼笔环肽-聚赖氨酸-金复合物处理可显示出标记的丝，但强度不够。