GCaMP6F Fluorescence in Ex Vivo Intestinal Preparation
Bryan Yoo, Jessica Griffiths, Sarkis Mazmanian
Published: 2021-11-03 DOI: 10.17504/protocols.io.bzqap5se
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Abstract
Protocol for GCaMP6F imaging in ex vivo intestinal tissue used in Yoo et al 2021
Steps
1.
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.
2.
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.
3.
C21 was added at 10nM and GCaMP6F fluorescence was detected on an upright microscope (Zeiss, Oberkochen, Germany-Examiner D1).
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