Feb 16, 2025

Public workspaceWhole Gut Transit Time, Fecal Water Content, and Fecal Output

 Forked from Whole Gut Transit Time, Fecal Water Content, and Fecal Output
DOI
dx.doi.org/10.17504/protocols.io.3byl4wxd2vo5/v1
  • 1Caltech
Chelsie Steele
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DOI: dx.doi.org/10.17504/protocols.io.3byl4wxd2vo5/v1
Protocol Citation: Chelsie Steele, Yujie Fan 2025. Whole Gut Transit Time, Fecal Water Content, and Fecal Output. protocols.io https://dx.doi.org/10.17504/protocols.io.3byl4wxd2vo5/v1
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: January 07, 2025
Last Modified: February 16, 2025
Protocol Integer ID: 117841
Funders Acknowledgements:
Aligning Science Across Parkinson’s
Grant ID: ASAP-020495
Abstract
Protocol for obtaining whole gut transit time
Experiment Preparation
Experiment Preparation
1d
1d
6% (w/v) carmine red (Sigma-Aldrich, St. Louis, MO) with 0.5% methylcellulose (Sigma-Aldrich)  was dissolved and autoclaved prior to use.
1d
Experiment
Experiment
Mice were orally gavaged with 150 µL of carmine red solution.
5m
Following gavage, mice were single-housed with no bedding for the duration of the experiment, and animals were not fasted beforehand.
Over the 3 hours following gavage, the mice were checked hourly, the number of stools counted, and three stools per animal were collected in pre-weighed, 1.5 mL microcentrifuge tube.
5h
Three hours after gavaging, each pellet collected was checked for the presence of carmine red, and the time of initial carmine red pellet expulsion was recorded as GI transit time.
If the mice did not have any carmine red present by three hours, their stool was checked every 15 minutes until presence of carmine red was seen and the time of initial carmine red pellet expulsion was recorded as GI transit time.
Sample Processing
Sample Processing
12h
12h
The mass of collected fecal pellets was determined, and pellets were left to dry in an 60 ºC oven overnight before weighing the desiccated pellets and calculating the pellets’ initial water content.
12h