BACKGROUND: The intestinal wall has a complex topographical architecture. The multi-layered network of the enteric nervous system and its intercellular interactions are difficult to map using traditional section-based or whole-mount histology. With the advent of optical clearing techniques, it has become feasible to visualize intact tissue and organs in 3D. However, as yet, a gap still needs to be filled in that no in-depth analysis has been performed yet on the potential of different clearing techniques for the small intestine. AIM: The goal of this study was to identify an optimal clearing protocol for in toto imaging of mouse intestinal tissue. METHODS: Five aqueous-based clearing protocols (SeeDB2, CUBIC, ScaleS, Ce3D, and UbasM) and four organic reagent-based clearing protocols (3DISCO, iDISCO+, uDISCO, and Visikol® ) were assessed in segments of small intestine from CX3CR1GFP/GFP and wild-type mice. Following clearing, optical transparency, tissue morphology, green fluorescent protein (GFP) fluorescence retention, and compatibility with (immuno-)labeling were analyzed. KEY RESULTS: All organic reagent-based clearing protocols-except for Visikol-rendered tissue highly transparent but led to substantial tissue shrinkage and deformation. Of the aqueous-based protocols, only Ce3D yielded full-thickness tissue transparency. In addition, Ce3D displayed excellent GFP retention and preservation of tissue morphology. CONCLUSIONS: Ce3D emerged as a most efficient protocol for enabling rapid full-thickness 3D mapping of the mouse intestinal wall.
BACKGROUND: The intestinal wall has a complex topographical architecture. The multi-layered network of the enteric nervous system and its intercellular interactions are difficult to map using traditional section-based or whole-mount histology. With the advent of optical clearing techniques, it has become feasible to visualize intact tissue and organs in 3D. However, as yet, a gap still needs to be filled in that no in-depth analysis has been performed yet on the potential of different clearing techniques for the small intestine. AIM: The goal of this study was to identify an optimal clearing protocol for in toto imaging of mouse intestinal tissue. METHODS: Five aqueous-based clearing protocols (SeeDB2, CUBIC, ScaleS, Ce3D, and UbasM) and four organic reagent-based clearing protocols (3DISCO, iDISCO+, uDISCO, and Visikol® ) were assessed in segments of small intestine from CX3CR1GFP/GFP and wild-type mice. Following clearing, optical transparency, tissue morphology, green fluorescent protein (GFP) fluorescence retention, and compatibility with (immuno-)labeling were analyzed. KEY RESULTS: All organic reagent-based clearing protocols-except for Visikol-rendered tissue highly transparent but led to substantial tissue shrinkage and deformation. Of the aqueous-based protocols, only Ce3D yielded full-thickness tissue transparency. In addition, Ce3D displayed excellent GFP retention and preservation of tissue morphology. CONCLUSIONS: Ce3D emerged as a most efficient protocol for enabling rapid full-thickness 3D mapping of the mouse intestinal wall.
Authors: David M Young; Siavash Fazel Darbandi; Grace Schwartz; Zachary Bonzell; Deniz Yuruk; Mai Nojima; Laurent C Gole; John Lr Rubenstein; Weimiao Yu; Stephan J Sanders Journal: Elife Date: 2021-02-11 Impact factor: 8.140
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Authors: Neil Marr; Mark Hopkinson; Andrew P Hibbert; Andrew A Pitsillides; Chavaunne T Thorpe Journal: Biol Proced Online Date: 2020-07-01 Impact factor: 3.244
Authors: Elina Nürnberg; Mario Vitacolonna; Julia Klicks; Elena von Molitor; Tiziana Cesetti; Florian Keller; Roman Bruch; Torsten Ertongur-Fauth; Katja Riedel; Paul Scholz; Thorsten Lau; Richard Schneider; Julia Meier; Mathias Hafner; Rüdiger Rudolf Journal: Front Mol Biosci Date: 2020-02-21
Authors: Manuela Gries; Anne Christmann; Steven Schulte; Maximilian Weyland; Stephanie Rommel; Monika Martin; Marko Baller; Ralph Röth; Stefanie Schmitteckert; Marcus Unger; Yang Liu; Frederik Sommer; Timo Mühlhaus; Michael Schroda; Jean-Pierre Timmermans; Isabel Pintelon; Gudrun A Rappold; Markus Britschgi; Hilal Lashuel; Michael D Menger; Matthias W Laschke; Beate Niesler; Karl-Herbert Schäfer Journal: Mol Neurodegener Date: 2021-06-02 Impact factor: 14.195