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PMID: 6409845 Published · ppublish English Journal Article

A simple post-embedding system for the rapid demonstration of tissue antigens under the electron microscope.

The Histochemical journal ·Vol. 15 ·No. 6 ·1983-06-00 ·Pages 543-55

Newman GR, Jasani B, Williams ED

Abstract

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.

MeSH Terms
Animals Humans Immunoglobulin lambda-Chains/metabolism Immunologic Techniques Insulin/metabolism Islets of Langerhans/ultrastructure Microscopy, Electron/methods Palatine Tonsil/ultrastructure Pancreas/ultrastructure Plastics Rats Thyrotropin/metabolism
Chemicals
Immunoglobulin lambda-Chains Insulin Plastics Thyrotropin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Newman G R
Jasani B
Williams E D
References (16)
16 references, click to expand
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Article Info
Journal
The Histochemical journal
Abbr.
Histochem J
ISSN
0018-2214
Published
1983-06-00
Pages
543-55
Language
English
Region
Netherlands
NLM ID
0163161
Subset
IM
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