Anat Biol Anthropol.  2022 Jun;35(2):57-66. 10.11637/aba.2022.35.2.57.

Deep Tissue Clearing for Three-dimensional Imaging Analysis of Murine Pancreas

Affiliations
  • 1Department of Anatomy, Kyungpook National University, School of Medicine, Korea
  • 2Bio-Medical Research Institute, Kyungpook National University Hospital, Korea
  • 3Immune Square Inc., Korea
  • 4Binaree, Inc., Korea
  • 5Biocenter, Gyeonggido Business & Science Accelerator, Korea
  • 6Graduate School of East-West Medical Science, Kyung Hee University, Korea
  • 7Deparment of Developmental and Cell Biology, University of California, Irvine, USA
  • 8Department of Artificial Intelligence & Big Data Engineering, Daegu Catholic University, Korea
  • 9Department of Internal Medicine, Chungbuk National University Hospital, Korea
  • 10Department of Internal Medicine, Chungbuk National University, College of Medicine, Korea
  • 11Clinical Omics Institute, Kyungpook National University, Korea
  • 12Department of Anatomy, Yonsei University College of Medicine, Korea

Abstract

Optical imaging of the vascular system is necessary for structural, functional, developmental, and pathological biology. Recent developments in optical tissue-clearing methods have significantly advanced large-scale tissue researches by integrating a three-dimensional (3D) visualization of topographical vasculature, so we aimed to enhance the vascular structure of the murine pancreas which are highly vascularized structure. In this study, the adult C57/BL6 murine pancreatic tissue was cleared after lectin staining, resulting in the reconstructed structures for 3D imaging within a short period without damaging. Enhancing the visualization processes using light-sheet fluorescence microscopy (LSFM) together with 3D reconstruction imaging tool, we studied in depth of the entire vascular system, which further enabled to reconstruct 3D structure of the vasculature in pancreas and islets. We successfully implemented the normal murine pancreatic tissues including exocrine and endocrine cells and vascular structures, reflecting pancreatic β-cell function. The size of islets of each pancreatic lobe were not different but the number of islets in each lobe was variable in our quantification. In conclusion, this deep tissue clearing technique for 3D imaging could be applied in future studies to further understand the topological interactions of various cells and blood vessels within intact organs and tissues.

Keyword

Light-sheet fluorescence microscopy; Pancreas; Three-dimensional; Tissue clearing; Vascular system
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