Diamine oxidase-gold labels histamine in human mast-cell granules: a new enzyme-affinity ultrastructural method.
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.
Dvorak
A M
AM
Department of Pathology, Beth Israel Hospital, Boston, Massachusetts 02215.
Morgan
E S
ES
Schleimer
R P
RP
Lichtenstein
L M
LM
D006631:Amine Oxidase (Copper-Containing) / Q000737:chemistry*; D002478:Cells, Cultured; D006046:Gold / Q000737:chemistry*; D006632:Histamine / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D008168:Lung / Q000166:cytology; D008407:Mast Cells / Q000378:metabolism* / Q000648:ultrastructure; D008854:Microscopy, Electron
D006046:Gold; D006632:Histamine; D006631:Amine Oxidase (Copper-Containing)