Establishing a Whole-Cell Configuration through the Patch Clamp Method

Begin with an immobilized mouse brain slice placed in a recording chamber of the electrophysiology setup filled with an artificial cerebrospinal fluid to maintain neuronal viability.

The setup contains a pre-assembled cationic solution-filled recording micropipette connected to an amplifier and a pressure control unit.

Under a microscope, locate a neuron and position the micropipette near it.

Apply positive air pressure to clear any debris.

Move the micropipette toward the neuron until a dimple forms on the membrane, indicating neuronal proximity.

Apply weak suction to pull a small membrane patch inside the micropipette.

This creates a tight seal that increases the micropipette resistance, confirming a stable seal.

Maintain the cell's potential at a physiological resting potential for cell stability.

Then, apply brief, strong suction to disrupt the patch membrane, establishing a direct connection with the cell's interior.

This whole-cell configuration is ready for recording neuronal electrical activity.

Using the fine focus wheel of the microscope, start focusing down while lowering the micropipette gradually. Always focus down first, and then, lower the micropipette to the plane o