Nov 02, 2021
GCaMP6F Fluorescence in Ex Vivo Intestinal Preparation
- Bryan Yoo1,
- Jessica Griffiths1,
- Sarkis Mazmanian1
- 1California Institute of Technology
- Mazmanian Lab
Protocol Citation: Bryan Yoo, Jessica Griffiths, Sarkis Mazmanian 2021. GCaMP6F Fluorescence in Ex Vivo Intestinal Preparation. protocols.io https://dx.doi.org/10.17504/protocols.io.bzqap5se
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: November 02, 2021
Last Modified: November 02, 2021
Protocol Integer ID: 54754
Funders Acknowledgements:
Center for Environmental and Microbial Interactions (CEMI)
Grant ID: n/a
Emerald Foundation
Grant ID: n/a
Heritage Medical Research Institute
Grant ID: n/a
Department of Defense
Grant ID: PD160030
National Institutes of Health
Grant ID: GM007616 and DK078938
Aligning Science Across Parkinson’s
Grant ID: ASAP-000375
Aligning Science Across Parkinson's
Grant ID: ASAP-020495
Abstract
Protocol for GCaMP6F imaging in ex vivo intestinal tissue used in Yoo et al 2021
Small intestinal tissue was quickly harvested from ChAT-Cre mice, flushed and placed in oxygenated (95% O2, 5% CO2), ice cold Krebs-Henseleit solution for 1 hour followed by 15 min at room temperature.
A segment was cut along the mesenteric attachment, and pinned flat (mucosa facing down) on a Sylgard-lined recording chamber (Warner Instruments, PH1) in oxygenated Krebs-Henseleit solution.
C21 was added at 10nM and GCaMP6F fluorescence was detected on an upright microscope (Zeiss, Oberkochen, Germany-Examiner D1).