Three-dimensional tooth imaging using multiphoton and second harmonic generation microscopy

Abstract

Detailed morphological and cellular information relating to the human tooth have traditionally been obtained through histological studies that required decalcification, staining, and fixation. With the recent invention of multiphoton microscopy, it has become possible to acquire high resolution images without histological procedures. Using an epiilluminated multiphoton microscope, we obtained two-photon excited autofluorescence and second harmonic generation (SHG) images of <i>ex vivo</i> human tooth. By combining these two imaging modalities we obtained submicron resolution images of the enamel, dentin, and the periodontal ligaments. The enamel emits endogenous two-photon autofluorescence. The structure of the dentin is visible from both the autofluorescence and second harmonic generation signals. The periodontal ligament composed mostly of collagen can be visualized by SHG imaging. We also constructed three dimensional images of the enamel, dentin, and periodontal ligament. The effectiveness of using multiphoton and second harmonic generation microscopy to obtain structural information of teeth suggest its potential use in dental diagnostics.