Demonstration of the cytoskeleton of lens epithelial cells with gold sol techniques.
1. 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.
Spindler
M
M
Institute of Biochemistry, Martin-Luther-University, Halle/Saale, Germany.
Iwig
M
M
D008841:Actin Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure; D000818:Animals; D001426:Bacterial Proteins; D046508:Culture Techniques; D003599:Cytoskeleton / Q000648:ultrastructure*; D004848:Epithelium / Q000648:ultrastructure; D007150:Immunohistochemistry; D007908:Lens, Crystalline / Q000648:ultrastructure*; D008854:Microscopy, Electron; D010590:Phalloidine; D011107:Polylysine; D019809:Streptavidin; D016707:Tissue Fixation
D001426:Bacterial Proteins; D010590:Phalloidine; D011107:Polylysine; D019809:Streptavidin