Optogenetic Electrophysiology Recording of Cells with Light-Dependent Ion Channels

Place a coverslip containing embryonic kidney cells expressing light-dependent ion channels and a fluorescent reporter protein on a silicon-applied measuring chamber. Close it and add an external buffer.

Position this chamber under a microscope. Place a bath electrode filled with agar. Perfuse with a buffer to remove residual media and dead cells.

Using a fluorescence filter, identify and focus on the target cell.

Attach a patch pipette to a micromanipulator. Apply pressure to prevent clogging and approach the cell.

Position the pipette near the cell and release positive pressure to form a membrane patch at the tip.

After forming the membrane patch, apply negative pressure to rupture the membrane.

Apply a light of a specific wavelength to open the ion channels. The movement of ions is recorded as voltage change by the electrode.