Paired Patch Clamp Recording of the Neuronal Connections in Acute Brain Slices

Secure a brain slice with the target region in a recording chamber.

Position a recording pipette containing an internal solution over a postsynaptic neuron and achieve a whole-cell configuration.

Introduce a searching patch pipette near a presynaptic neuron.

This pipette is filled with a sodium-based solution to prevent neuron depolarization while searching for the presynaptic cell.

Use gentle suction to establish a loose-seal configuration with the searching pipette.

Hold the membrane potential and apply large current pulses to trigger an action potential in the presynaptic neuron.

Monitor the postsynaptic response to this stimulation.

If no response is detected, patch another presynaptic neuron and repeat stimulation.

Continue this process until a response is observed in the postsynaptic neuron.

Replace the searching pipette with a recording pipette and establish a whole-cell configuration in the presynaptic neuron.

Apply current pulses to elicit action potentials in the presynaptic neuron and record the postsynaptic response.

After preparing slices of the target region, allow them to incubate. During this time, pull the patch pipettes, which should have a long and slender shank.