METHOD FOR ENHANCEMENT OF ELECTRON MICROSCOPIC VISUALIZATION OF EMBEDDED ANTIGEN BY BRIDGING OSMIUM TO URANIUM ANTIBODY WITH THIOCARBOHYDRAZIDE

Thiocarbohydrazide (TCH), a bidentate ligand, capable of bridging osmium to osmium in ultrathin sections (OTO method), was shown to possess the additional property of binding osmium to uranium-labeled antibody. On sections in which acrolein-fixed antigen had been localized for electron microscopy by...

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Veröffentlicht in:J. Histochem. Cytochem., 14: 711-18(Oct. 1966) 14: 711-18(Oct. 1966), 1966-10, Vol.14 (10), p.711-718
Hauptverfasser: STERNBERGER, LUDWIG A, HANKER, JACOB S, DONATI, EDWARD J, PETRALI, JOHN P, SELIGMAN, ARNOLD M
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Sprache:eng
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Zusammenfassung:Thiocarbohydrazide (TCH), a bidentate ligand, capable of bridging osmium to osmium in ultrathin sections (OTO method), was shown to possess the additional property of binding osmium to uranium-labeled antibody. On sections in which acrolein-fixed antigen had been localized for electron microscopy by uranium antibody, application of TCH followed by osmication (T-O procedure) greatly enhanced the specific contrast afforded. In addition, the T-O procedure yielded better resolution and better stability to the electron beam than the immunouranium technique employed without this intensification. Even osmium tetroxide-fixed tissue could be used, providing thiosemicarbazide (TSC) was employed to block the later reaction of TCH with tissue-bound osmium and to limit, thereby, the ligation of TCH to the uranium-labeled antibody.
ISSN:0022-1554
1551-5044
DOI:10.1177/14.10.711