Three-Dimensional Analysis of the Human Pancreas

Endocrinology. 2018 Mar 1;159(3):1393-1400. doi: 10.1210/en.2017-03076.

Abstract

Pancreatic islets are endocrine micro-organs scattered throughout the exocrine pancreas. Islets are surrounded by a network of vasculature, ducts, neurons, and extracellular matrix. Three-dimensional imaging is critical for such structural analyses. We have adapted transparent tissue tomography to develop a method to image thick pancreatic tissue slices (1 mm) with multifluorescent channels. This method takes only 2 to 3 days from specimen preparation and immunohistochemical staining to clearing tissues and imaging. Reconstruction of the intact pancreas visualizes islets with β, α, and δ cells together with their surrounding networks. Capturing several hundred islets at once ensures sufficient power for statistical analyses. Further surface rendering provides clear views of the anatomical relationship between islets and their microenvironment as well as the basis for volumetric quantification. As a proof-of-principle demonstration, we show an islet size-dependent increase of intraislet capillary density and an inverse decrease in sphericity.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Arterioles / diagnostic imaging
  • Capillaries / diagnostic imaging
  • Frozen Sections
  • Glucagon / analysis
  • Humans
  • Imaging, Three-Dimensional / methods*
  • Immunohistochemistry
  • Insulin / analysis
  • Islets of Langerhans / blood supply
  • Islets of Langerhans / chemistry
  • Islets of Langerhans / diagnostic imaging*
  • Mice
  • Pancreas / chemistry
  • Pancreas / diagnostic imaging*
  • Platelet Endothelial Cell Adhesion Molecule-1 / analysis
  • Somatostatin / analysis

Substances

  • Antibodies, Monoclonal
  • Insulin
  • Platelet Endothelial Cell Adhesion Molecule-1
  • Somatostatin
  • Glucagon