Mapping Large Extensions of Flat Dentin Through Digital Microscopy: Introduction to the Method and Possible Applications

Purpose: To introduce a mapping method to characterize large dentin surfaces using digital microscopy and to discuss the advantages and possible applications of the method. Materials and Methods: Twenty unerupted third molars were sectioned transversally exposing coronal dentin surfaces. The microscopic mosaic method was used to generate a large field image with the resolution necessary to measure characteristics of dentin tubules. The AxioVision 4.7 software was used to control a motorized optical microscope and the process of acquiring approximately 400 small images to generate each dentin mosaic. An image analysis routine measured the number of tubules (NT) and the ratio between the total area of tubules and the area of the mosaic - the area fraction (AF) - of each mosaic. An automatic procedure transformed the mosaic image into a color map, providing a direct visual representation of tubule density through colors. The dentin maps were used for a comparative qualitative analysis of tubule density distribution of each sample. Results: The results for NT (92450 to 196029 tubules/sample) and AF (4.12% to 11.10%) demonstrated a wide variation among dentin samples. The maps confirmed the microstructure variety, also revealing strong local variations in tubule density within each sample. Conclusion: The mapping method was able to perform dentin morphology characterization and is a valuable tool for producing a baseline for dentin adhesion studies. The method could be also useful in determining the real contribution of dentin structures to the final adhesion quality. Keywords: adhesion, digital microscopy, image analysis, dentin map