Summary
To obtain contrast-staining of glycogen in electron microscopy, various contrast-enhancing additives can be used in combination with potassium osmate. Examples are potassium ferrocyanide and certain nitrogen heterocyclic compounds, such as triazoles. In the reaction sequence leading to contrast-stained glycogen, a primary reaction is the formation of glycogen osmate. This reaction was studied with isolated glycogen. On the basis of the stoichiometric findings, a molecular structure of the reaction product is proposed; apparently, osmate is bound by glycogen because of the presence of suitably located hydroxyl groups. The resulting compound is not itself sufficiently electron dense, but it binds 1,2,4-triazole as an additional ligand. Secondary reactions can result in additional osmium binding, and finally to osmium (IV) deposits, leading to contrast.
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Riemersma, J.C., Alsbach, E.J.J. & de Bruijn, W.C. Chemical aspects of glycogen contrast-staining by potassium osmate. Histochem J 16, 123–136 (1984). https://doi.org/10.1007/BF01003544
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DOI: https://doi.org/10.1007/BF01003544