The aim of this work was to investigate relationship between refractive index (n) and mineral content (MC) (vol %) of enamel and dentin using swept-source optical coherence tomography (SS-OCT) and transverse microradiography (TMR). Enamel and dentin blocks were partitioned into three regions. The middle partition of each sample was covered with a nail polish to protect the sound area during exposure to the treatment solutions. Samples were demineralized in a demineralizing solution, which was refreshed once a week, for 2 months. One window was covered with acid-resistant varnish, leaving the other window exposed; the samples were placed in a solution for remineralization. Samples then were sliced into disks with thickness of 300 μm to 400 μm and placed on metal plate in order to capture cross-sectional images of sound, demineralized and remineralized regions by OCT at 1319 nm center wavelength. The n then was calculated via formula using image analysis software. Following n measurement, these specimens were further polished for the TMR analysis. Correlation between OCT n and TMR MC was examined. A significant and highly positive correlation was found between the measured n and the actual MC at the corresponding locations (Pearson correlation coefficients (r) were 0.94 and 0.97 in enamel and 0.95 and 0.91 in dentin after de-/remineralization process, respectively p < 0.05). OCT showed a potential for quantitative analysis of the mineral loss or gain by measuring of the n in vitro. Supported by the grant from the Japanese Ministry of Education, Global Center of Excellence (GCOE) Program, "International Research Center for Molecular Science in Tooth and Bone Diseases.".