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Studies on reaction and binding of monoamines after fixation and processing for electron microscopy with special reference to fixation with potassium permanganate

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In the present paper certain properties of potassium permanganate (KMnO4), a fixative used for electron microscopical investigations, have been studied in model test tube experiments and on tissues. Evidence was obtained that KMnO4 reacts with different types of biogenic monoamines resulting in a formation of a precipitate. In addition, also various monoamine analogues, precursors and metabolites reacts with KMnO4. The reaction taking place may be an oxidation-reduction-reaction in which KMnO4 is reduced, probably mainly to manganese dioxide by hydroxyl groups of the amines and related compounds. This is corroborated by the fact that no reaction takes place between KMnO4 and β-phenylethylamine or amphetamine, two substances, which lack hydroxyl groups.

Using labelled monoamines evidence was obtained that the amine partly is retained within the precipitate formed after the reaction with KMnO4 and also in tissues fixed with KMnO4, indicating a possibility to perform autoradiographic studies on KMnO4 fixed tissue.

Electron microscopic studies on tissues fixed under various conditions revealed that fixation with low concentrations (0.6 and 1.0%) of KMnO4 and at high temperatures (about 20° C) leads to inferior results as to general morphology and as to the visualization of intraneuronal amine stores.

Different types of permanganates were tested as fixatives. These results show that fixation with permanganates with monovalent metallic ions (K+, Li+ and Na+) give good results of comparable quality, whereas fixation with zinc permanganate results in seriously destroyed tissues. However, tissue fixed with calcium permanganate reveals very distinct membranes. Furthermore, evidence was obtained that fixation with high concentrations of LiMnO4 (6 and 9%) and NaMnO4 (6 and 9%) was more sensitive as to the demonstration of monoamines at the ultrastructural level as compared to 3% KMnO4. Thus, with e.g. 6 and 9% LiMnO4 small granular vesicles could be seen in slices from the caudate nucleus after incubation with α-methyl-dopamine. This was not possible when using 3% KMnO4 as a fixative.

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  1. Department of Histology, Karolinska Institutet, Stockholm, Sweden

    T. Hökfelt & G. Jonsson

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  1. T. Hökfelt
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Hökfelt, T., Jonsson, G. Studies on reaction and binding of monoamines after fixation and processing for electron microscopy with special reference to fixation with potassium permanganate. Histochemie 16, 45–67 (1968). https://doi.org/10.1007/BF00306211

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