Real-Time Calcium Imaging of a Mouse Medial Prefrontal Cortex Using a Miniscope

Take an anesthetized transduced mouse implanted with a gradient-index lens targeting the medial prefrontal cortex, where neurons express genetically encoded calcium indicators.

Remove the lens cap and clean its surface for clear light transmission.

Secure the miniscope to the lens holder.

Adjust the miniscope position to identify the optimal focal plane. This adjustment ensures clear visualization of neuronal cell bodies.

Detach the cable and allow the mouse to recover.

Connect the cable to the miniscope and transfer the mouse to a chamber with a behavioral recording setup.

During imaging, the miniscope emits light to excite the calcium indicators, which fluoresce upon intracellular calcium influx.

The gradient-index lens relays these optical signals from deep brain tissues to the miniscope, reducing light scattering and enabling real-time visualization of neuronal activity.

Process the recorded calcium transient activity with behavioral observations to correlate neuronal activity to specific behaviors.

After briefly anesthetizing the mouse with isoflurane, loosen the locking screw in the base with a small screwdriver before removing the protective cap, and clean the surface of the GRIN le