Microspectrofluorometry in biogenic amine research

Abstract

Microspectrofluorometry has played an important part in the development and understanding of fluorescence techniques for the demonstration of biogenic amines. This paper reviews briefly the techniques currently available for the characterization and localization of biogenic amines, and considers the basic design considerations applicable to the construction of a microspectrofluorometer, with special emphasis on the automated correction of spectral data. Spectral data for the commonly occurring biogenic amines dopamine, noradrenaline and 5-hydroxytryptamine have been summarized and the criteria applicable to the spectral analysis of the individual amines and their mixtures discussed. Discrepancies between spectral data derived from tissues and model systems are reviewed and emphasis is given to the importance of biochemical studies of amines prior to definitive microspectrofluorometric analysis. The conceptual basis of quantitative microspectrofluorometry is reviewed with a particular emphasis on the limitations of such studies. The analysis of terminal fields by scanning microspectrofluorometry and its application to the study of extracellular amine distribution is discussed.