Establishing a Whole-Cell Voltage-Clamp Configuration for Electrophysiological Recordings in Brain Slices
Transcript
Take a recording pipette with a wire electrode in an electrolyte solution.
Position the pipette above an immobilized brain slice.
Adjust the pipette current to reduce background signals.
Use a microscope to locate the target neuron and move the pipette toward the neuron.
Apply a slight positive pressure and lock it.
The positive pressure disturbs the tissue surface, indicating proximity to the cell.
Bring the pipette closer until a dimple forms on the membrane, signifying correct positioning.
Release the positive pressure.
Observe an increase in resistance as the cell forms a seal with the pipette.
Hold the membrane potential at a constant negative voltage for stability.
Pull the micropipette away to remove excess pressure.
Adjust the pipette's fast and slow electrical responses for accuracy.
Apply brief suction to break through the cell, establishing a direct electrical connection.
The whole-cell voltage clamp configuration is now ready for electrophysiology recording.
