Optical Coherence Tomography was used to monitor subsurface caries evolution process in vitro. Human tooth was used and bacteria were employed to induce caries lesions. Twenty-five human third molars, were used in this study. The teeth were cut longitudinally at mesio-distal direction; the surfaces were coated with nail varnish except for two squared windows (2x4 mm); at the cement-enamel junction. Artificial lesions were induced by a S. Mutans microbiological culture. The samples (N = 50) were divided into groups according to the demineralization time: 3, 5, 7, 9 and 11 days. The culture medium, was changed each 48 hours. After the demineralization process the samples were rinsed with double-deionized water and stored in a humid environment. The OCT system was implemented with average power of 96 μW in the sample arm, providing a 23 μm of axial resolution. The images were produced with lateral scans step of 10 μm. The detection system was composed by a detector, a demodulator and a computer. With the images generated by OCT it was possible to determine the lesion depth as function of sample exposition time to microbiological culture. We observed that the depth of the lesion in the root dentine increased from 70 μm to 230 μm, depending of exposure time, and follows the bacterial population growth law. This OCT system accurately depicts hard dental tissue and it was able to detect early caries in its structure, providing a powerful contactless high resolution image of lesions.