Establishing a Whole-Cell Configuration through the Patch Clamp Method
Transkript
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.
