Optical clearing improves the imaging depth and signal-to-noise ratio for digital analysis and three-dimensional projection of the human enteric nervous system.

Neurogastroenterology and Motility : the Official Journal of the European Gastrointestinal Motility Society
Y A LiuS C Tang

Abstract

Due to the dispersed nature of neurites and fibers, the microtome-based 2-dimensional histology provides only a limited perspective of the enteric nervous system. To visualize the enteric plexus, we applied optical clearing to avoid scattering in the human ileum to facilitate photon penetration for 3-dimensional (3-D) microscopy of the neural tissue. Human ileal specimens were derived by trimming the donor bowel due to its excess length during the clinical trial of small intestinal transplantation. The pan-neuronal marker PGP9.5 was used as the immunostaining target to reveal the enteric plexuses. The labeled tissues were immersed in the optical-clearing solution prior to deep-tissue confocal microscopy. The serial sections were digitally analyzed and processed by reconstruction algorithms for 3-D visualization. Optical clearing of the ileal specimen led to less fluorescence signal decay along the focal path in the tissue and a higher signal-to-noise ratio of the confocal micrographs in comparison with the untreated saline control. Taking advantage of the high signal-to-noise ratio images, we applied software-based signal analysis to identify the presence of the nerve fibers and quantify the signal peaks. The image stacks deriv...Continue Reading

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Citations

Jun 27, 2013·Diabetologia·Shiue-Cheng TangYa-Yuan Fu
Mar 17, 2012·American Journal of Physiology. Gastrointestinal and Liver Physiology·Yuan-An LiuShiue-Cheng Tang
Sep 10, 2014·Diabetes, Obesity & Metabolism·S-C TangH-J Chien
Mar 19, 2013·Neurogastroenterology and Motility : the Official Journal of the European Gastrointestinal Motility Society·Y A LiuS C Tang
Oct 6, 2011·Nature Reviews. Gastroenterology & Hepatology·Katherine Smith
Feb 15, 2019·Neurogastroenterology and Motility : the Official Journal of the European Gastrointestinal Motility Society·Gleison D P BossolaniJean-Pierre Timmermans
Jan 16, 2014·American Journal of Physiology. Endocrinology and Metabolism·Jyuhn-Huarng JuangShiue-Cheng Tang
May 19, 2020·American Journal of Physiology. Gastrointestinal and Liver Physiology·Cambrian Y Liu, D Brent Polk
Sep 12, 2019·American Journal of Physiology. Gastrointestinal and Liver Physiology·Hung-Jen ChienShiue-Cheng Tang
Oct 23, 2012·American Journal of Physiology. Gastrointestinal and Liver Physiology·Ya-Yuan FuShiue-Cheng Tang
Apr 27, 2021·American Journal of Physiology. Endocrinology and Metabolism·Chien-Chia ChenShiue-Cheng Tang

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