Proper treatment of dental caries demands detection of carious lesions at an early stage and a minimal invasive cavity preparation to preserve the maximum tooth structure. Various devices use fluorescence for caries detection via recording the red fluorescence generated by dentin caries under illumination.The aim of this study was to evaluate the porphyrin and pentosidine involvement in the red fluorescence observed in enamel and dentin caries when illuminated with the Soprolife® camera (Sopro, Acteon Group, La Ciotat, France) and Vistacam® camera (Dürr Dental AG, Bietigheim-Bissingen, Germany). Three techniques were used: single photon fluorescence spectroscopy, micro-Raman spectroscopy, and color analysis with ImajeJ software. Cross-sections of human teeth, scored from 0-6 with the International Caries Detection and Assessment System (ICDAS), were examined by fluorescence microscopy. Teeth spectra of each ICDAS score were compared with those of protoporphyrin IX (PpIX), porphyrin I, and pentosidine solutions. A specific confocal Raman microscopy analysis was realized and a Red-Green-Blue model analysis of Soprolife® images was performed using ImageJ software to compare the color variations on ICDAS score 1 and 2. Fluorescence spectroscopy and micro- Raman spectroscopy revealed the presence of PpIX in carious enamel and dentin. The clinical relevance of this experimentation was that the increased knowledge of the fluorescence aids for caries detection could improve the preventive approach, thus reducing the operative one.