Diamine oxidase-gold labels histamine in human mast-cell granules: A new enzyme-affinity ultrastructural method

A. M. Dvorak*, E. S. Morgan, R. P. Schleimer, L. M. Lichtenstein

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

30 Scopus citations

Abstract

We developed a post-embedding ultrastructural enzyme affinity-gold technique to label histamine. Diamine oxidase (DAO)-gold complex was prepared and tested for specificity with routinely processed, Epon-embedded, histamine-rich human mast-cell granules and histamine-agar test blocks, both of which were labeled. Specificity controls included removal of staining by DAO digestion of Epon sections containing mast cells or by filtering DAO- gold over histamine-agarose beads, and a wide variety of controls (for these specificity controls) that did not abrogate mast-cell granule or histamine- agar block labeling with DAO-gold. Internal negative controls in each sample included the failure of DAO-gold to stain Type II alveolar pneumocyte lamellar bodies or altered, swollen mast-cell granules from which histamine has been released. The new post-embedding enzyme affinity-gold technique is effective in optimally prepared ultrastructural samples that allow clear identification of key ultrastructural morphology.

Original languageEnglish (US)
Pages (from-to)787-800
Number of pages14
JournalJournal of Histochemistry and Cytochemistry
Volume41
Issue number6
DOIs
StatePublished - 1993

Keywords

  • Diamine oxidase
  • Histaminase
  • Histamine
  • Mast cells
  • Secretory granules
  • Type II alveolar pneumocytes
  • Ultrastructural enzyme affinity-gold method

ASJC Scopus subject areas

  • Anatomy
  • Histology

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