Optical clearing and 3D analysis optimized for mouse and human pancreata

Alexandra Alvarsson, Maria Jimenez-Gonzalez, Rosemary Li, Carolina Rosselot, Nikolaos Tzavaras, Zhuhao Wu, Sarah A. Stanley

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
Article numbere4103
JournalBio-protocol
Volume11
Issue number15
DOIs
StatePublished - 5 Aug 2021

Keywords

  • Clearing
  • IDISCO
  • Innervation
  • Light sheet
  • Pancreas

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