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

David Wynford-Thomas, Bharat Jasani, Geoffrey R. Newman

Research output: Contribution to journalArticlepeer-review

28 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)387-396
Number of pages10
JournalThe Histochemical Journal
Volume18
Issue number7
DOIs
Publication statusPublished - Jul 1 1986

ASJC Scopus subject areas

  • Anatomy
  • Cell Biology

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