This in vitro study investigated the prevalence, location, and pathway of the second mesiobuccal canal (MB-2) in 45 first and second maxillary molars using the operating microscope (OM). Initially location and negotiation of MB-2 were attempted without magnification. Teeth in which MB-2 was not located or could not be negotiated were further explored under OM. Roots where MB-2 could not be negotiated even with OM were cross-sectioned and inspected microscopically. Morphometric measurements were performed to map the location of MB-2. Without magnification an apparent MB-2 orifice was located in 42 teeth and the canal negotiated in 31 (69%). With OM one additional apparent MB-2 orifice was located, and five previously identified canals were negotiated (total 80%). The root cross-sections confirmed the absence of MB-2 in all nine teeth where it was not negotiated. Location of MB-2 varied randomly. In conclusion MB-2 can be negotiated in 80% of maxillary molars, although an orifice is apparent in 96% of the teeth. Ability to negotiate MB-2 is facilitated by OM.

Objective: The objective of this study was to evaluate and compare the two scanning electron microscope (SEM) preparation protocols and determine the better SEM preparation technique to study stem cells on human amniotic membrane (hAM) scaffold. Materials and Methods: Formaldehyde-based protocol and glutaraldehyde-based protocol were compared to evaluate the quality of SEM images for stem cells cultured on hAM scaffold. Results: The results suggested that formaldehyde-based protocol is better than glutaraldehyde-based protocol in terms of showing clearer topography of the membrane as well as the boarders of the cells. To provide intact surface of the SEM sample and avoid possible ruptures of the hAM or the thin cell layer, it is recommended to perform the dehydration step using graded alcohol concentrations of 20%, 30%, 40%, 50%, 60%, 70%, 80%, and 90%, one time for each and twice in 100% for 10 min each. Gold sputter-coating step is not recommended as it does not improve the image quality. Conclusions: To obtain clear SEM images, it is recommended to run a preliminary study to determine the better chemicals and conditions of sample preparation even when following preexisting protocols.