To begin, implant the recording electrodes and inject the relevant viruses in the mouse skull. After anesthetizing the mice, connect the optical fibers sequentially in the order of the recording site before inserting the head stage. Then, set the 635 nanometer light pulse with a 10%duty cycle at 20 hertz, and set up the light stimulation for 10 seconds during recording.
Set the intensity at the optical fiber tips at three milliwatts. Next, use a fiber photometry system to record Gcamp6m signals throughout the behavior testing. Simultaneously, record calcium signals, local field potentials, and electroencephalogram under optogenetic stimulation, along with corresponding behavioral performances.
Finally, employ transistor-transistor logic signal synchronization tagging to ensure temporal consistency in recorded data. Brief optogenetic stimulation elicited robust after discharges, along with remarkable increases in calcium activities in the target brain regions. A 30 hertz response was observed in the opto-stimulating regio, induced by the activation of optogenetic proteins via blue light LED.