@article{014de8d1efe641898d1e9595e0f7972d,
title = "Optical clearing and 3D analysis optimized for mouse and human pancreata",
abstract = "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.",
keywords = "Clearing, IDISCO, Innervation, Light sheet, Pancreas",
author = "Alexandra Alvarsson and Maria Jimenez-Gonzalez and Rosemary Li and Carolina Rosselot and Nikolaos Tzavaras and Zhuhao Wu and Stanley, {Sarah A.}",
note = "Funding Information: This protocol was adapted from the previous study published by Alvarsson et al. (2020). A.A. was supported by a senior postdoctoral fellowship from the Charles H. Revson Foundation (grant no. 18-25), a fellowship from Sweden-America Foundation (Ernst O Eks fond), and a postdoctoral scholarship from the Swedish Society for Medical Research (SSMF). M.J.G and A.A. are supported by the Department of Defense Discovery Award (# W81XWH-20-1-0156). This work was also supported by the American Diabetes Association Pathway to Stop Diabetes Grant ADA #1-17-ACE-31 and, in part, by grants from the NIH (R01NS097184, OT2OD024912, and R01DK124461), Department of Defense (W81XWH-20-1-0345), and the Alexander and Alexandrine Sinsheimer Scholar Award. This work was supported in part by a Mindich Child Health and Development Institute Pilot and Feasibility Grant and by a Mount Sinai Distinguished Scholar award. Microscopy and image analysis were performed at the Microscopy CoRE at the Icahn School of Medicine at Mount Sinai. Publisher Copyright: Copyright {\textcopyright} 2021 The Authors.",
year = "2021",
month = aug,
day = "5",
doi = "10.21769/BioProtoc.4103",
language = "English",
volume = "11",
journal = "Bio-protocol",
issn = "2331-8325",
publisher = "Bio-protocol LLC",
number = "15",
}