A simple post-embedding system for the rapid demonstration of tissue antigens under the electron microscope.
A simple and versatile technique for the preparation of ultra-thin sections, which can be stained immunohistochemically directly on electron microscope grids, is presented. An anti-hapten immunoperoxidase procedure has been adapted for use on tissue fixed in a purified monomeric glutaraldehyde--picric acid mixture, and embedded in 'L R White', a recently formulated plastic resin. This plastic tolerates the use of partial dehydration of tissue, resulting in higher antigenic yields. In addition, no etching of ultra-thin sections is necessary, and the whole immunostaining procedure can be completed in less than 2 h. A comparison of commonly used fixatives is discussed. High-resolution micrographs showing general staining (uranyl acetate--lead citrate) of rat pancreas, and immunostaining of insulin and TSH in storage granules in perfusion-fixed rat tissue and of lambda-chain immunoreactive cells in immersion-fixed human tonsil are included as examples.
D000818:Animals; D006801:Humans; D007146:Immunoglobulin lambda-Chains / Q000378:metabolism; D007158:Immunologic Techniques; D007328:Insulin / Q000378:metabolism; D007515:Islets of Langerhans / Q000648:ultrastructure; D008854:Microscopy, Electron / Q000379:methods*; D014066:Palatine Tonsil / Q000648:ultrastructure; D010179:Pancreas / Q000648:ultrastructure; D010969:Plastics; D051381:Rats; D013972:Thyrotropin / Q000378:metabolism
D007146:Immunoglobulin lambda-Chains; D007328:Insulin; D010969:Plastics; D013972:Thyrotropin