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Immunogold technique for electron microscopic detection of a bacterial surface antigen in ultrathin sections.
A method for the immunocytochemical detection of a bacterial surface antigen was developed using the postembedding immunogold technique (PPID) in ultrathin sections of semithin Epon-embedded Epon sections. For this purpose, sections were blocked with 1% bovine serum albumin (BSA), rinsed in phosphate-buffered saline (PBS), and incubated with a monoclonal antibody to treponema pallidum antigens. Next, sections were incubated with goat anti-mouse immunoglobulins coupled to colloidal gold particles (6 nm) followed by the application of a silver enhancement procedure. Treponemal structures were easily detected on the sections, and the localization of the antigen was verified by back-contrasting in the dark using iodine-ethylenediaminetetraacetic acid. This technique was used for the localization of the surface antigen in ultrathin sections and will be particularly useful in studies of treponemal morphology.Q:
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