Endocrinology
Endocrinology
5 years ago

Technical Resource article (METHODS AND TOOLS).

Steve Seung-Young Lee, Scott K Olehnik, Zachary C Wesner, Jonas L Fowler, Stephen J Kron, Manami Hara
Pancreatic islets are endocrine micro-organs scattered throughout the exocrine pancreas. Islets are surrounded by a network of vasculature, ducts, neurons and extracellular matrix. Three-dimensional imaging is critical for such structural analyses. We have adapted Tissue Transparent Tomography (T3) to develop a method to image thick pancreatic tissue slices (1 mm) with multi-fluorescent channels. This method takes only 2-3 days from specimen preparation and immunohistochemical staining to clearing tissues and imaging. Reconstruction of the intact pancreas visualizes islets with beta-, alpha- and delta-cells together with their surrounding networks. Capturing several hundred islets at once ensures sufficient power for statistical analyses. Further surface rendering provides clear views of the anatomical relationship between islets and their microenvironment as well as the basis for volumetric quantification. As a proof-of-principle demonstration, we show an islet-size dependent increase of intra-islet capillary density and an inverse decrease in sphericity.

Publisher URL: http://doi.org/10.1210/en.2017-03076

DOI: 10.1210/en.2017-03076

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