Histotomography of the odontoblast processes at the dentine–enamel junction of permanent healthy human teeth in the confocal laser scanning microscope

Abstract

The translucency of teeth allows the non-destructive subsurface visualisation of their microstructure by confocal laser scanning microscopy (CLSM) at a level of about 150 μm below the surface. The dentine–enamel junction (DEJ) is accessible only directly adjacent to the cervix of the tooth. Therefore teeth have to be sectioned for studying marginal areas of the dental hard tissue. The potential of the technique for (pseudo) three-dimensional visualisation allows the study of an array of individual confocal images, the interpretation of which is similar to that of macroscopic tomographs (CT-scan, MRI). Additionally, the extended focus mode yields the overlay of individual confocal images in the form of a two-dimensional projection. This mode of operation proved to be particularly suited for the visualisation of odontoblast processes in their whole extension. The three-dimensional junction between enamel and dentine, the branches of the odontoblast processes and their interactions with the DEJ is demonstrable by CLSM without staining or other procedures of sample preparation. The direct microscopic comparison between samples, either fresh or kept in a humid chamber, and Technovit-embedded sample blocks gives evidence that the risk of artefacts by sample storage or by the embedding procedure is minimal. The tomographs limited to subsurface areas of the tissue also exclude mechanical surface artefacts due to grinding or cutting.