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- Start by placing a recording chamber containing a sample in a bath solution, whose composition matches the physiological extracellular environment or the cytoplasm, onto a microscope stage. Advance the recording micropipette, a pipette containing a silver electrode bathed in an electrolyte solution, which mimics the intracellular conditions, toward the neuron by applying positive pressure, and make contact with the cell membrane.
Gently switch from positive to negative pressure to form a tight seal between the membrane and the pipette. Set a holding potential on the amplifier to maintain the cell at a constant voltage. Then rupture the cell membrane within the micropipette by applying suction.
Once the cell membrane ruptures, the holding potential causes the flow of ions through voltage-gated ion channels, creating a membrane potential, which is recorded by the recording electrode. Immediately record the membrane potential. The recording electrode transmits the signal to the feedback amplifier, which subtracts the membrane potential from the holding potential-- an oscilloscope, which presents a visual display of the membrane potential, and a computer. In the following protocol, w
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