An intravital microscopy-based approach to assess intestinal permeability and epithelial cell shedding performance

Martínez-Sánchez, Luz DC., Pradhan, Rashmita, Ngo, Phuong A., Erkert, Lena, Becker, Lukas S., Watson, Alastair J., Atreya, Imke, Neurath, Markus F. and López-Posadas, Rocío (2020) An intravital microscopy-based approach to assess intestinal permeability and epithelial cell shedding performance. Journal of Visualized Experiments, 2020 (166). ISSN 1940-087X

[img] PDF (Accepted_Manuscript) - Accepted Version
Restricted to Repository staff only until 3 December 2022.

Download (539kB) | Request a copy

Abstract

Intravital microscopy of the gut using confocal imaging allows real time observation of epithelial cell shedding and barrier leakage in living animals. Therefore, the intestinal mucosa of anesthetized mice is topically stained with unspecific staining (acriflavine) and a fluorescent tracer (rhodamine-B dextran), mounted on a saline solution-rinsed plate and directly imaged using a confocal microscope. This technique can complement other non-invasive techniques to identify leakage of intestinal permeability, such as transmucosal passage of orally administered tracers. Besides this, the approach presented here allows the direct observation of cell shedding events at real-time. In combination with appropriate fluorescent reporter mice, this approach is suitable for shedding light into cellular and molecular mechanisms controlling intestinal epithelial cell extrusion, as well as to other biological processes. In the last decades, interesting studies using intravital microscopy have contributed to knowledge on endothelial permeability, immune cell gut homing, immune-epithelial communication and invasion of luminal components, among others. Together, the protocol presented here would not only help increase the understanding of mechanisms controlling epithelial cell extrusion, but could also be the basis for the developmental of other approaches to be used as instruments to visualize other highly dynamic cellular process, even in other tissues. Among technical limitations, optical properties of the specific tissue, as well as the selected imaging technology and microscope configuration, would in turn, determine the imaging working distance, and resolution of acquired images.

Item Type: Article
Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
Related URLs:
Depositing User: LivePure Connector
Date Deposited: 30 Jan 2021 01:11
Last Modified: 12 Jun 2021 12:00
URI: https://ueaeprints.uea.ac.uk/id/eprint/78329
DOI: 10.3791/60790

Actions (login required)

View Item View Item