Potential clamp of isolated dialyzed neuron: minimalization of the effect of series resistance.
A modification of the technique of intracellular dialysis of isolated single excitable cells, such as rat spinal ganglion neuron, suitable for potential clamping of its somatic membrane is described. The advantage of the new modification is the substantial reduction of the effect of inherent resistance in series (RS) to the membrane resistance (RM) on precision of potential clamping. This is attained by reversal of cell position in the perfusion pipette resulting in an approximately tenfold reduction in the area of active membrane. The resistance of this area proportionally increased while RS remained unchanged. Hence the error in potential fixation, which is inversely proportional to the ratio RM/RS, is by approximately one order smaller with the new technique than with the original one. An essential step in the new technique is the osmotic expansion of the cell to improve the contact of the cell with the perfusion pipette in the pore and to facilitate disruption of the appropriate part of the membrane. All features and advantages of the technique of intracellular dialysis, such as simplicity, the possibility to easily change ionic composition of media, and/or to apply drugs to any side of the membrane in the same cell, etc., have been maintained.
Stolc
S
S
Centre of Physiological Sciences, Slovak Academy of Sciences, Bratislava.
Nemcek
V
V
Boska
D
D
D002462:Cell Membrane / Q000502:physiology; D003956:Dialysis; D004594:Electrophysiology; D006863:Hydrogen-Ion Concentration; D008564:Membrane Potentials; D009474:Neurons / Q000502:physiology; D009997:Osmotic Pressure