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Protocol
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Human Pancreas Optical Clearing by iDISCO and VISIKOL

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Description
Optical clearing of tissues to improve deep tissue microscopy was described over a century ago and methods have improved considerably following publication of the CLARITY method from Karl Deisseroth in 2013. Improvements in organic and inorganic methods are constantly reported for multiple species and organs. Large volume imaging was needed for human pancreas studies due to known heterogeneity in normal islet sizes and proportions of endocrine cells. Lobularity in islet beta-cell loss and islet inflammation is also seen in patients with type 1 diabetes. The iDISCO protocol allows for rapid clearing, easy staining, and high resolution confocal imaging. Human pancreas samples were cleared using iDISCO as originally published. This protocol describes steps to clear 4mm x 6mm x 400um deep sections of 4% paraformaldehyde fixed human pancreas using iDISCO-like reagents purchased from Visikol.
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