Apr 24, 2024

Public workspaceA protocol for tissue clearing and three-dimensional imaging of human sigmoid mucosal biopsies V.2

A protocol for tissue clearing and three-dimensional imaging of human sigmoid mucosal biopsies
  • 1University of California at Los Angeles (UCLA)
Open access
Protocol CitationPu-Qing Yuan, Tao Li, Yvette Taché 2024. A protocol for tissue clearing and three-dimensional imaging of human sigmoid mucosal biopsies . protocols.io https://dx.doi.org/10.17504/protocols.io.6qpvr8dkblmk/v2Version created by Pu-Qing Yuan
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: April 24, 2024
Last Modified: April 24, 2024
Protocol Integer ID: 98739
Keywords: Tissue clearing, 3D imaging, human sigmoid mucosal biopsy, nerve fibers
Funders Acknowledgement:
NIH/SPARC
Grant ID: 1OT2OD024899-01
Abstract
This protocol was developed for tissue clearing and 3D imaging of human sigmoid mucosal biopsies by adapting and modifying the original CLARITY tissue clearing technique/immunofluorescence protocols that we established for 3D imaging and phenotyping of human colonic enteric nervous system. By using this new protocol, a piece of entire human sigmoid mucosal biopsy specimen with the volume about 6x3x1-2 mm (length x wide x thickness) can be completely cleared within 3-4 days. The 3D images and videos generated from cleared biopsy samples showed clear spatial views of nerve innervation, distributions of mast cells and enteric glial cells, and configurations of mast cells with nerve fibers in the sigmoid colonic mucosa.
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