Optical Clearing and 3D Analysis Optimized for Mouse and Human Pancreata

The pancreas is a heavily innervated organ, but pancreatic innervation can be challenging to comprehensively assess using conventional histological methods. However, recent advances in whole-mount tissue clearing and 3D rendering techniques have allowed detailed reconstructions of pancreatic innerva...

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Bibliographic Details
Published inBio-protocol Vol. 11; no. 15; p. e4103
Main Authors Alvarsson, Alexandra, Jimenez-Gonzalez, Maria, Li, Rosemary, Rosselot, Carolina, Tzavaras, Nikolaos, Wu, Zhuhao, Stanley, Sarah A
Format Journal Article
LanguageEnglish
Published United States Bio-Protocol 05.08.2021
Bio-protocol LLC
Subjects
Biology
Methods
Clearing
Pancreas
iDISCO
Innervation
Light sheet
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Summary:The pancreas is a heavily innervated organ, but pancreatic innervation can be challenging to comprehensively assess using conventional histological methods. However, recent advances in whole-mount tissue clearing and 3D rendering techniques have allowed detailed reconstructions of pancreatic innervation. Optical clearing is used to enhance tissue transparency and reduce light scattering, thus eliminating the need to section the tissue. Here, we describe a modified version of the optical tissue clearing protocol iDISCO+ (immunolabeling-enabled three-dimensional imaging of solvent-cleared organs) optimized for pancreatic innervation and endocrine markers. The protocol takes 13-19 days, depending on tissue size. In addition, we include protocols for imaging using light sheet and confocal microscopes and for 3D segmentation of pancreatic innervation and endocrine cells using Imaris.
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ISSN:2331-8325
2331-8325
DOI:10.21769/BioProtoc.4103