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Establishing a Whole-Cell Configuration through the Patch Clamp Method

Establishing a Whole-Cell Configuration through the Patch Clamp Method

Transcript

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.

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