Immunohistochemical localization of cell surface receptors using a novel method permitting simple, rapid and reliable LM/EM correlation

A method is presented which allows correlative serial section analysis by light and electron microscopy of cell surface antigens in monolayer cultures. Sites of antigenicity are shown by deposition of diaminobenzidine after pre-embedding, immunoperoxidase immunocytochemistry. Osmication is replaced...

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Bibliographic Details
Published in:The Histochemical journal 1986-07, Vol.18 (7), p.387-396
Main Authors: WYNFORD-THOMAS, D, BHARAT JASANI, NEWMAN, G. R
Format: Article
Language:English
Subjects:
Antigens, Surface - analysis
Biological and medical sciences
Cell Line
Cell Membrane - ultrastructure
Cell receptors
Cell structures and functions
ErbB Receptors - analysis
Fundamental and applied biological sciences. Psychology
Histological Techniques
Humans
Immunoenzyme Techniques
Microscopy, Electron
Molecular and cellular biology
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Summary:A method is presented which allows correlative serial section analysis by light and electron microscopy of cell surface antigens in monolayer cultures. Sites of antigenicity are shown by deposition of diaminobenzidine after pre-embedding, immunoperoxidase immunocytochemistry. Osmication is replaced by the use of gold chloride which specifically enhances the electron density of diaminobenzidine. In addition gold chloride bound to diaminobenzidine survives embedding and provides the basis for a post-embedding photochemical amplification method. Immunostained cells are embedded in LR White by a rapid technique which preserves their structure and leaves them available for subsequent post-embedding immunocytochemistry. The method is illustrated by the demonstration of epidermal growth factor (EGF) receptors on the EGF receptor-rich human carcinoma cell line A431 using a well characterized monoclonal antibody raised against EGF receptor.
ISSN:0018-2214
1573-6865
DOI:10.1007/BF01675220