Ultrastructural identification of peripheral myelin proteins by a pre‐embedding immunogold labeling method

Abstract

Ultrastructural immunolabeling of peripheral nervous system components is an important tool to study the relation between structure and function. Owing to the scarcity of certain antigens and the dense structure of the peripheral nerve, a pre-embedding technique is likely appropriate. After several investigations on procedures for pre-embedding immunolabeling, we propose a method that offers a good compromise between detection of antigenic sites and preservation of morphology at the ultrastructural level, and that is easy to use and suitable for investigations on peripheral nerve biopsies from humans. Pre-fixation by immersion in paraformaldehyde/glutaraldehyde is necessary to stabilize the ultrastructure. Then, ultrasmall gold particles with silver enhancement are advised. Antibodies against myelin protein zero and myelin basic protein were chosen for demonstration. The same technique was applied to localize a 35 kDa myelin protein.

Antibody-functionalized polydiacetylene coatings on nanoporous membranes for microorganism detection

“…The coatings were polymerized and then fixed in 0.1 M sodium phosphate buffer with 4% formalin at pH 7.4. Immuno-TEM observations were performed following the method of Canron et al, 10 by Paragon Bioservices, Inc. in Baltimore, MD. Briefly, the coatings were exposed to anti-bovine antibodies labelled with biotin, exposed to streptavidin-labeled 6 nm gold particles, embedded, sectioned, stained and observed.…”