The polarizing images of hard tissues including bone and cementum show characteristic features of different birefringence fortheir microstructures. Nevertheless, the clear mechanism of the amplified birefringence under polarizing microscope has not been well understood. We hypothesized that the unique polarized light could be accumulated in the microtubules due to the decreased refractory angle by the inside lower-density matrix, and then the accumulated light in the microtubules could be dispersed brightly. In order to elucidate the polarizing effect on the microtubules, the dentinal tubules in different conditions were observed, and compared with each other to explain their birefringence phenomena. In the decalcified sections of normal tooth, the dentinal tubules located near the pulp chamber showed strong birefringence, while the sclerosed dentinal tubules near the dentino-enamel junction did not show the birefringence. The birefringence was more conspicuous in the longitudinal sectionsof dentinal tubules than in the cross sections. In the decalcified sections of complex odontoma, which produced abnormal and immature dentinal tubules, the birefringence was not observed in the shrunken dentinal tubules filled with dense materials, while the peritubular matrix showed clear birefringence. The birefringence was also observed in the collagen fibers in the connective tissue, and continuously strong in the immature cemental materials containing precollagen fibers. However, the highly mineralized osteodentine did not show the birefringence. Taken together, these data suggest that the microtubules composed ofless-dense matrix than the background tissue, i.e., dentinal tubules, Haversian canals, etc., produce the amplified birefringence by the polarizing light according to the hypothesis of microtubule refraction.