Immunolabeling of Cleared Human Pancreata Provides Insights into Three-dimensional Pancreatic Anatomy and Pathology.

Immunolabeling of Cleared Human Pancreata Provides Insights into Three-dimensional Pancreatic Anatomy and Pathology.

Am J Pathol. 2018 Apr 20;:

Authors: Noë M, Rezaee N, Asrani K, Skaro M, Groot VP, Wu PH, Olson MT, Hong SM, Kim SJ, Weiss MJ, Wolfgang CL, Makary MA, He J, Cameron JL, Wirtz D, Roberts NJ, Offerhaus GJA, Brosens LAA, Wood LD, Hruban RH

Abstract
Visualizing pathologies in three dimensions (3D) can provide unique insights into the biology of human diseases. A rapid and easy-to-implement dibenzyl ether-based technique was used to clear thick sections of surgically resected human pancreatic parenchyma. Protocols were applicable to both fresh and formalin-fixed paraffin-embedded tissue. The penetration of antibodies into dense pancreatic parenchyma was optimized using both gradually increasing antibody concentrations and centrifugal flow. Immunolabeling with antibodies against CK19 was visualized using both light sheet and confocal laser scanning microscopy. The technique was successfully applied to 26 sections of pancreas, providing 3D images of normal pancreatic tissue, pancreatic intraepithelial neoplasia, intraductal papillary mucinous neoplasms, and infiltrating pancreatic ductal adenocarcinomas. 3D visualization highlighted processes which are hard to conceptualize in two dimensions, such as invasive carcinoma growing into what appeared to be pre-existing pancreatic ducts and within venules, and the tracking of long cords of neoplastic cells parallel to blood vessels. Expanding this technique to formalin-fixed, paraffin-embedded tissue opens pathology archives to 3D visualization of unique biosamples and rare diseases. The application of immunolabeling and clearing to human pancreatic parenchyma provides detailed visualization of normal pancreatic anatomy, and can be used to characterize the 3D architecture of diseases including pancreatic intraepithelial neoplasia, intraductal papillary mucinous neoplasm, and pancreatic ductal adenocarcinomas.

PMID: 29684363 [PubMed - as supplied by publisher]

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