Jan 16, 2024
GCaMP6f imaging using a mini-epifluorescence microscope (Mendonça et al 2024)
- Marcelo D Mendonça1,2,3
- 1Champalimaud Research, Champalimaud Foundation, Lisbon 1400-038, Portugal;
- 2Champalimaud Clinical Centre, Champalimaud Foundation, Lisbon 1400-038, Portugal;
- 3NOVA Medical School | Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Lisbon 1169-056, Portugal
Protocol Citation: Marcelo D Mendonça 2024. GCaMP6f imaging using a mini-epifluorescence microscope (Mendonça et al 2024). protocols.io https://dx.doi.org/10.17504/protocols.io.kqdg3xdx7g25/v1
Manuscript citation:
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 12, 2024
Last Modified: May 31, 2024
Protocol Integer ID: 93468
Keywords: ASAPCRN
Abstract
GCaMP6f imaging using a mini-epifluorescence microscope methods from Mendonça et al 2024.
Guidelines
This procedure excludes surgical procedures that would occur prior to the imaging day. Please see 'virus injections and lens placement procedure' for those details.
Fluorescence images were acquired at 10 Hz and the LED power was set 40-60% with a gain of 4
Before start
Mice are briefly anesthetized using a mixture of isoflurane and oxygen (1% isoflurane at 1L/min) and the mini-epifluorescence microscope was attached to the baseplate.
GCaMP6f imaging
GCaMP6f imaging
Once the baseplate (that is attached to the mouse) is attached to the mini-epifluorescence microscope, mice are recovered in their home cage for 15-20 minutes prior to starting imaging.
Image acquisition parameters were always set to the same parameters between sessions to be able to compare the activity recorded.
With the baseplate attached to the microscope, mice are them imaged during the FR4/1s task in 3 performance days. (see Single-limb fast FR4 operant task methods for details)