Confocal fluorescence microscopy images of dentinal porosity at the dentin-enamel junction UM_0001_C1_2_DEJ

Confocal image stacks were acquired with a Leica TCS SP8 microscope, using a 40x/1.3NA oil immersion lens. A wavelength of 561 nm (LED laser DPSS 561) was used to excite Rhodamine B fluorescence which was collected in a range of 567-636 nm with a hybrid detector. The acquisitions were performed on the polished surface of crown dentin up to 900 µm from the DEJ in the direction of the pulp (cf Fig.1a-b). The scanning was performed using 26-52 µW laser power with a linear increase in depth to compensate for scattering and absorption with a pixel dwell time of 300 ns and a line average of 3 acquisitions. The scan consisted of a mosaic of 5 overlapping stacks of 205.5×205.5 µm2 images with a pixel size of 100 nm, collected over 20.65 µm in depth with 350 nm sampling, forming a 934.4×205.5×20.65 µm3 volume. The resulting experimental dataset was thus 2055×9344×59 voxels in shape with 100×100×350 nm3 voxel size.