Qualitative evaluation of scanning electron microscopy methods in a study of the resin cement/dentine adhesive...

Sample preparation and imaging techniques for scanning electron microscopy (SEM) of dehydrated dental samples can hinder the structural analyses. This study qualitatively evaluated images obtained with two different protocols of SEM preparation and analysis to assess the dentine adhesive interface. The crown and root dentine of 12 bovine incisors were subjected to cementation with the resin cement RelyX U100 or RelyX ARC/SBMP (n = 6). After storage for 7 days in a moist environment at 37 ± 1°C, the dentine samples were dehydrated in an ascending alcohol series, and three specimens from each group were coated with gold or carbon and examined in a high-vacuum (JEOL JSM-6360LV, 10 kV) or low-vacuum (FEI Quanta 200F, 15-30 kV) microscope. Images were obtained at magnifications between 50 and 2,000×, but with different working distances. The use of high vacuum for carbon and gold coating and SEM visualization led to cracks in the samples. A small number of cracks can be described in the specimens subjected to the low-vacuum technique. The protocol for SEM imaging in low vacuum was considered more appropriate for preservation of the integrity of the evaluated structures.