Abstract
Abstract The pancreas is a unique organ where the endocrine microorgans are embedded in the exocrine tissue. To visualize the spatial distribution of pancreatic cells in situ also with three dimensions observation could provide a better understanding of the overall organization of endocrine cells and the relationship between different structures in the pancreas. In this research, we apply an optical-clearing solution, FocusClear, to minimize the random light scattering in the mouse pancreas and show the overall spatial organization of β-cells and islets in the pancreas. This technique, with the advantage than conventional fluorescence microscopy and alleviates technical difficulties such as collection and operating for serial histological sections and avoids structural defects during the sectioning process. Our results thus establish the basis for a new platform which is a noninvasive three-dimensional microscopic imaging method for in situ visualization and assessment of pancreatic islet microstructure. The three-dimensional images of the fluorescence-labeled structure. Specifically, the islet vasculature and sympathetic nerve fibers were labeled by vessel painting (lipophilic dye labeling of blood vessels) and immunofluorescence staining of neural markers, respectively, for confocal microscopy. Optical sectioning and reconstruction of bulk tissue are simple and rapid. Volume and number of β-cells within a block of tissue can be automatically quantified by freely available software.