Source Paper
High-fat diet disrupts a septal control on feeding to promote obesity in male mice
Jiang S, Lai S, Jing H, Wu X, Li F et al.
Nat Commun • 2025
Single-cell calcium imaging during food consumption
Objective: Single-cell calcium imaging to record neuronal activity in the lateral septum (LS) during food consumption in freely moving mice
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Protocol Steps
Viral injection and GRIN lens implantation
Inject 200 nl of AAV2/9-hEF1a-DIO-GCaMP6s into the LS at coordinates AP: +0.5 mm, ML: ±0.45 mm, DV: –2.8 mm at 60 nl/min injection rate. Insert GRIN lens 100 μm above injection site and secure with dental cement.
View evidence from paper
“inject 200 nl of AAV virus solution into the LS at a slow rate (60 nl/min)”
Recovery period
Allow mice to recover for at least 3 weeks after viral injection and GRIN lens implantation.
View evidence from paper
“at least three weeks prior to any manipulations to allow for viral infection”
Baseplate attachment
After 4-6 weeks of GCaMP6s injection, fix a baseplate that matches the miniscope to each mouse's skull with dental cement.
View evidence from paper
“After 4-6 weeks of GCaMP6s injection, a baseplate that matched the miniscope was fixed”
Adaptive training
Provide mice with 10-minute adaptive training sessions for at least 3 days before imaging to habituate them to the experimental setup.
View evidence from paper
“mice received 10-minute adaptive training for at least 3 days”
Calcium imaging during food consumption
Randomly place food pellets for freely moving mice and simultaneously record calcium imaging data during food consumption. Include at least 10 food intake periods per imaging session.
View evidence from paper
“There were at least 10 food intake periods during the whole imaging session”
Data acquisition
Acquire imaging data at 30-Hz frame rate using UCLA Miniscope-DAQ-DT-Software.
View evidence from paper
“Imaging data were acquired at a 30-Hz frame rate and collected using UCLA Miniscope-DAQ-DT-Software”
Signal processing
Process calcium signals using CNMF-E software to extract motion-corrected GCaMP6s fluorescence dynamics from individual neurons. Quantify as Z-scores or ΔF/F values with baselines defined as mean fluorescence during first 2 seconds of each trial.
View evidence from paper
“Calcium signal processing was performed using CNMF-E software to extract motion-corrected GCaMP6s fluorescence dynamics”