A chemical mechanism for tissue staining by osmium tetroxide-ferrocyanide mixtures
Abstract
The presence of Fe(CN)6(-4) provides sequential, one-electron reduction pathways for OSO4. An equilibrium is established containing OSO4, Fe(CN)6(-4), Fe(CN)6(-3), OSO2(OH)4(-4), and labile cyano-bridged OS-Fe species containing Os in nominal oxidation states of VIII, VII, and VI. These osmium complexes are chelated by appropriately placed donor atoms in the macromolecular tissue matrix, and chelation facilitates the reduction of osmium in situ to lower oxidation states (predominantly IV) that are relatively nonlabile. The greater reactivity and concentration of the Os(VII and VI) intermediates in this system leads to more Os deposition than OsO4 alone; the chelation is responsible for the immobilization of Os and the observed staining pattern in electron micrographs. Chemical data from model systems and electron micrographs of tissue are presented in support of this mechanism.
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Potassium ferrocyanide in combination with osmium tetroxide enhances the visualization of cellular membranes.
“Later, the introduction of potassium ferrocyanide (potassium hexacyanoferrate (II); K4[Fe(CN)6]; Karnovsky, 1971; White et al., 1979; McDonald, 1984) or potassium ferricyanide (potassium ferricyanide (III); K3[Fe(CN)6]; Rivlin and Raymond, 1987) in combination with osmium tetroxide was used to enhance the visualization of cellular membranes, as well as certain aspects of cell morphology.”
From Unambiguous identification of asymmetric and symmetric synapses using volume electron microscopy (2024) · cites this paper as White et al., 1979
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Cited by 108
- Authors
- White, D. L., Mazurkiewicz, J. E., & Barrnett, R. J.
- ORCID
- White, D L
- Venue
- Journal of Histochemistry & Cytochemistry
- Publisher
- SAGE Publications
- Published
- 1979-07
- Volume
- 27
- Issue
- 7
- Pages
- 1084-1091
- ISSN
- 0022-1554 (ISSN-L)
- 1551-5044
- OpenAlex
- W2026975428
- Cited by
- 108
- References
- 0
- Citations by year
- 2026: 1 · 2025: 3 · 2024: 1 · 2023: 3 · 2022: 1 · 2021: 1 · 2020: 4 · 2019: 5 · 2017: 3 · 2016: 3 · 2015: 4 · 2013: 1 · 2012: 3
- FWCI
- 4.71
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Histochemistry & Cytochemistry (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Advanced Electron Microscopy Techniques and Applications
- Health Sciences › Medicine › Pharmacology › Biological Stains and Phytochemicals
- Health Sciences › Medicine › Anatomy › Medical and Biological Sciences
- Keywords
- osmium tetroxide, ferrocyanide, tissue staining, osmium complexes, one-electron reduction, oxidation state, chelation, staining mechanism, electron microscopy contrast
- MeSH terms
- Animals, Erythrocyte Membrane, Erythrocytes, Ferrocyanides, Histocytochemistry, Microscopy, Electron, Muscles, Osmium, Osmium Tetroxide, Sarcoplasmic Reticulum, Staining and Labeling, Rats
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