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The glyoxylic acid fluorescence histochemical method: a detailed account of the methodology for the visualization of central catecholamine neurons

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Summary

This paper gives a detailed description of the glyoxylic acid fluorescence histochemical method as designed for the highly sensitive visualization of catecholamine neurons. In this method, the primary catecholamines, dopamine and noradrenaline, are efficiently converted to intensely fluorescent 2-carboxymethyl-dihydroisoquinoline derivatives in a well defined reaction with glyoxylic acid. The method is carried out on sections from fresh or glyoxylic acid-perfused tissue, which are immersed in a glyoxylic acid solution, dried, and then reacted either by heating at +100°C, or by glyoxylic acid vapour treatment at +100°C. The method has a high reproducibility, is rapid and convenient, and if desired, sections of good quality can be ready for fluorescence microscopy within half an hour after the sacrifice of the animal.

The glyoxylic acid method demonstrates central and peripheral dopamine- and noradrenaline-containing neurons with an extraordinary sensitivity and precision. The entire adrenergic neuron, including the non-terminal portions of the axon and sometimes also the dendrites, becomes fluorescent, making the method ideal for neuroanatomical tracing of central catecholamine pathways. The spectral characteristics of the glyoxylic acid-induced fluorophores have been investigated, and it is concluded that the catecholamine fluorophores can be identified and distinguished by microspectrofluorometry from those of other fluorogenic monoamines known to occur in the vertebrate brain.

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  1. Department of Histology, University of Lund, Lund, Sweden

    Olle Lindvall & Anders Björklund

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  1. Olle Lindvall
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  2. Anders Björklund
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Lindvall, O., Björklund, A. The glyoxylic acid fluorescence histochemical method: a detailed account of the methodology for the visualization of central catecholamine neurons. Histochemistry 39, 97–127 (1974). https://doi.org/10.1007/BF00492041

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