ObjectivesThe fluorescence-aided identification technique (FIT) is based on the fluorescence properties of dental materials, specifically the intensity of their fluorescence compared to the autofluorescence of hard dental substances; this creates a perceived contrast between dental material and tooth. However, no studies to date have determined the extent to which the fluorescence intensity of tooth-colored dental materials must differ from that of natural autofluorescence to ensure reliable visual detection. The aim of this study was therefore to determine, for the first time, how pronounced the difference between fluorescence intensity and autofluorescence must be to reliably identify tooth-colored material. MethodsTen dentists assessed six different resin-based composite (RBC) samples of varying fluorescence intensity placed in the cavities of ten extracted teeth under standardized fluorescence-exciting illumination. The outcome variable was fluorescence perceptibility. Their assessments of the outcome variable were compared with measurements of the fluorescence intensities of the RBCs and the surrounding dental hard tissues, which were expressed as a fluorescence intensity ratio. Demographic data of the participants, including age, gender, and professional experience, were also recorded. ResultsNo significant differences were found for visual fluorescence perceptibility in relation to the explanatory variables of gender (p = 0.14), age (p = 0.13), and professional experience (p = 0.34). In contrast, the fluorescence intensity ratio was significantly different (p < 0.0001). SignificanceFor both clinicians and manufacturers, fluorescence intensity levels are important when selecting or developing FIT-compatible materials. Our results suggest that the fluorescence intensity levels of dental materials should be no more than 75% and no less than 200% of tooth natural autofluorescence to ensure reliable detection of tooth-colored materials.