In the present study, we investigated, using micro-Raman spectroscopy (Raman) and transverse microradiography, the influence of bicarbonate [sodium hydrogen carbonate (NaHCO3)] on the effects of carbonate ions in the mineral phase during demineralization (acid resistance test) of subsurface lesions. Baseline lesions were created by demineralizing bovine enamel, and specimens were then exposed to remineralization solutions containing 0, 5, or 50 mM bicarbonate. Acid resistance tests were performed on remineralized and sound enamel specimens. Raman spectra showed that carbonate and phosphate were incorporated into both surface layers and lesion bodies during remineralization in the presence of bicarbonate. Moreover, the presence of bicarbonate did not affect the rates of remineralization, although the average mineral profiles of remineralized enamel differed from those of sound enamel after acid resistance tests. Raman analyses enabled close evaluation of site-specific characteristics of carbonate and phosphate in subsurface lesions. In conclusion, incorporation of carbonate and phosphate ions into enamel subsurface lesions during remineralization does not affect the magnitude of remineralization or acid resistance.