The aluminum-formaldehyde (ALFA) histofluorescence method for improved visualization of catecholamines and indoleamines. 1. A detailed account of methodology for central nervous tissue using paraffin, cryostat or vibratome sections

Abstract

This present paper presents a new aluminum-formaldehyde (ALFA) histofluorescence method for highly sensitive visualization of central monoamine-containing neurons, based on perfusion with or immersion in buffers containing high concentrations of aluminum ions. Our previous studies have shown that perfusion or immersion of tissues with solutions containing high concentrations of magnesium results in an improvement in the visualization of intraneuronal catecholamines in the reaction with formaldehyde and glyoxylic acid. This study demonstrates that aluminum is considerably more efficient as a fluorescence-promoting agent, thus causing a further increase in the sensitivity of the formaldehyde method. Detailed protocols are given for the ALFA-method applied to paraffin sections of freeze-dried tissue, and to cryostat and Vibratome sections. The present ALFA technique applied to paraffin sections of freeze-dried tissue visualizes all known catecholamine neuron systems with a sensitivity comparable to, and for certain noradrenergic systems higher than, that of the previously published glyoxylic acid-Vibratome method. Furthermore, the use of freeze-dried, paraffin embedded tissue makes possible convenient storage and parallel processing of many specimens. This mode of processing also allows en bloc reaction, which is the only way by which consistent and reproducible fluorescence yields can be obtained throughout large series of sections and parallelly processed specimens. In animals pretreated with l-tryptophanand MAO-inhibitor the technique is also useful for studies on central indoleamine-containing systems in freeze-dried tissue. The ALFA procedure applied to cryostat and Vibratome sections gives a more sensitive and reproducible visualization of central catecholamine neurons than previous methods.