Ferric ion, ferrocyanide, and inorganic phosphate as cytochemical reactants at peripheral nodes of Ranvier

Ferric ion (Fe3+) and ferrocyanide (Fe(CN)64-) were used under a variety of conditions to stain nodes of Ranvier in mammalian peripheral nerves. Principal findings are: 1. Ferric ion will bind to the extracellular nodal gap substance if nerves are pretreated with a phosphate buffer; or, it will bind to the cytoplasmic surface of the nodal axolemma if pretreatment is with cacodylate or veronal--acetate buffer. 2. Ferrocyanide will bind to the inner surface of the nodal axolemma, where it may react with ferric ion to form a blue stain, or with an osmium compound to form a black stain. 3. Ferric ion and ferrocyanide are bound to nodes as colloidal precipitates, and may migrate away from their sites of formation. 4. Not all nodes in a single piece of tissue, or in a single fibre have identical staining properties. It is concluded that ferric ion, ferrocyanide, and inorganic phosphate are valuable as cytochemical reactants for peripheral nodes of Ranvier, but they must be used in carefully controlled experimental situations in order to avoid spurious results.