3D ultrastructural analysis of pancreatic beta cells by serial block face scanning electron microscopy

Authors

• MA Aronova
• EL McBride
• A Rao
• Q He
• G Zhang
• T Cai
• H Xu
• AL Notkins
• RD Leapman

Abstract

We have applied serial block-face scanning electron microscopy (SBF-SEM) to measure parameters that describe the architecture of pancreatic islets of Langerhans, microscopic endocrine organs that secrete insulin and glucagon for control of blood glucose. By analyzing entire mouse islets, it is possible to determine the numbers and total volume of insulin-containing beta granules with an approach that combines stereological measurements on 2D slices and full 3D volume measurements of whole cells. Results from an NOD mouse model for Type 1 diabetes have been compared with results from wild-type mice. We have also analyzed the organization of mitochondria in beta cells, since it has been suggested that alterations in mitochondrial fission and fusion might play a role in nutrient-induced apoptosis, with possible involvement in the pathophysiology of diabetes. Mitochondria are highly dynamic organelles that change their shape between discrete structures and large interconnected networks by selective fission and fusion of their membranes, resulting in fused networks can permeate through an entire cell and whose function is not fully understood. We have used SBF-SEM to quantify the connectivity of mitochondria in beta cells of pancreatic islets fixed at given time points.

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