Establishing a Whole-Cell Voltage-Clamp Configuration for Electrophysiological Recordings in Brain Slices
Transcrição
To obtain whole-cell configuration, place the recording pipette just above the slice, and offset pipette current in the Voltage Clamp mode. Apply slight positive pressure to the pipette and lock the stopcock.
Next, select a healthy cell with an intact membrane and approach the tissue with the pipette. The positive pressure should cause a slight disturbance on the tissue. Slowly bring the pipette closer to the cell in a diagonal motion until a small dimple is formed on the cell surface. Then, release the positive pressure lock. The cell will begin to form a seal and the resistance will increase above one gigaohm. In voltage clamp, hold the cell at -68 millivolts.
Subsequently, slightly pull the pipette away from the cell diagonally to remove excess pressure. Compensate for the fast and slow pipette capacitance. Apply a brief suction through the tube connected to the pipette holder to break through the cell and obtain a whole-cell configuration.
