The diagnosis of root caries, in particular the judgment of the activity of a visually observed lesion, is difficult. Quantitative determination of lesion severity would allow the lesion to be monitored with time, so that an indication of lesion activity could be obtained. This paper describes a step in the development of a method that provides such a quantitative determination. Specifically, fluorescein sodium salt is used as a penetrating dye, the subject of study being the relationship between dye concentration and porosity in demineralized root dentin. Fourteen human third molars were demineralized in vitro (lactic acid CMC-gel, pH 5; in each of 6 groups for 4, 7, 11, 14, 18, and 21 days). Fluorescein sodium salt (0.2 g/L) was applied for 2 min. Thin slices (+/- 130 microns) were cut from the root surfaces without water cooling. The dye fluorescence radiance in the demineralized dentin was determined by means of a micro-Raman spectroscope and compared with the mineral loss profiles measured with transverse microradiography (TMR). The TMR data were corrected for the difference in measurement area between the two measurement systems. Corrected TMR profiles were compared with the corresponding fluorescence scans, showing linear correspondence. The correlation coefficient was r = 0.96. We conclude that, after uptake of fluorescein sodium salt for 2 min, the dye concentration in an artificially produced root-surface caries lesion is proportional to the amount of mineral lost from that lesion.