Some Aspects of Instrumentation and Methods as Applied to Fluorometry at the Microscale

Abstract

During the last ten years fluorometry has become a well-known and very important tool in the field of quantitative cytochemistry, whereas biochemists had already developed this technique into an extremely valuable methodology ten to twenty years before. With regard to the quantitative staining reactions involved, cytofluorometry has evolved predominantly from the fluorescence microscopy of cells and tissues. Of course, the experience gained in the biochemists’ development of fluorometric methodology must also be utilized for the quantitation of fluorescence microscopy; no doubt, the use of fluorescence parameters to derive quantitative information in cytochemistry requires the same careful consideration of the physicochemical laws of fluorescence phenomena as that needed in chemistry and biochemistry. Thus, fluorometry of single cells and cellular compartments should profit by the considerable amount of experience that is available in the field of macrofluorometry with regard to instrumentation, methodology, and interpretation of results.