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Visualization and quantification of healthy and carious dentin structure using confocal laser scanning microscopy
Abstract
In this study, a fluorescence technique was developed for visualization of dentin using confocal laser scanning microscopy (CLSM). Eighteen extracted human teeth were used: 13 showing no clinical signs of caries and 5 with visually apparent decay. Preliminary study: All teeth were horizontally sectioned to approx. 200 μm thickness and pre-treated as follows: no pretreatment; vacuum only; ultrasonication only; sodium hypochlorite (NaOCl) only; vacuum and NaOCl; ultrasonication and NaOCl; or vacuum, ultrasonication and NaOCl. Samples were stained with Rhodamine 123 fluorescent dye at a concentration of 10-5 M in phosphate buffer saline for 1 to 24 hours. Caries study: Dentin surfaces, some with pre-existing caries, were visualized using CLSM. Most dentin tubules in sound dentin appeared open using CLSM, but most dentin tubules in carious dentin appeared closed or narrowed. Surface images obtained using CLSM were similar to those seen by SEM, but additional subsurface imaging was possible using CLSM at depth intervals of 1 μm to a depth of 30 - 50 μm. This technique shows good potential for non-invasive surface and subsurface imaging of dentin structures.
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