Some limitations of the cell-attached patch clamp technique: a two-electrode analysis.
With two independent patch electrodes sealed to small clusters of electrically coupled chick embryo cardiac cells, we have measured four parameters: true seal and patch resistance, channel conductance, and membrane potential. One electrode was in the cell-attached mode, and recorded current flowing in parallel through the membrane patch and seal. The second electrode, sealed on a different cell in the cluster, was in the whole cell recording configuration, and served to record or control the membrane potential of the cluster. We fit the four measured parameters to a simple electrical model to reveal errors not usually recognized in the patch-clamp technique. Among these are the following: (1) The apparent seal resistance, determined by changing the potential in a patch electrode, may be a poor estimate of true seal resistance, since it includes the parallel combination of seal- and patch-resistance. (2) Patch resistance may be influenced by the electrode filling solution, and is often much lower than is usually assumed. (3) With a small cell preparation that has an input resistance in the gigaohm range, measurements of single-channel conductance using a cell-attached patch electrode may be inaccurate because cell membrane potential does not remain constant as electrode potential is varied.
Fischmeister
R
R
Ayer
R K
RK
DeHaan
R L
RL
D000818:Animals; D002642:Chick Embryo; D046508:Culture Techniques; D004553:Electric Conductivity; D004594:Electrophysiology / Q000379:methods*; D006321:Heart / Q000196:embryology*; D007473:Ion Channels / Q000502:physiology; D008564:Membrane Potentials
D007473:Ion Channels