Discontinuous single electrode voltage-clamp measurements: assessment of clamp accuracy in Vicia faba guard cells.
The use of a discontinuous single electrode voltage-clamp (dSEVC) offers an attractive alternative to the patch-clamp technique, since whole-cell measurements can be performed with a single sharp electrode. Comparison of current-voltage relations, however, revealed a weaker voltage dependence of channels measured with the dSEVC compared to patch clamp. The accuracy of the dSEVC was tested on Vicia faba guard cells impaled with double-barrelled electrodes. The actual clamp potential was measured independently of the dSEVC, at the second barrel. The weaker voltage dependence of ion channels appeared to be due to an overestimation of the clamp potential by the dSEVC. The deviation between the intended and actual clamp potential showed a linear relationship with the injected current; on average a 126 mV deviation was found for a clamp current of 1 nA. The deviation was probably caused by a slow settling capacity at the electrode, not compensated by the dSEVC amplifier. It is concluded that the dSEVC method in its current state is only suited for the study of small ion conductances in plant cells.
Roelfsema
M R
MR
Julius-von-Sachs-Institut für Biowisssenschaften, Lehrstuhl für Molekulare Pflanzenphysiologie und Biophysik, Universität Würzburg, Julius-von-Sachs-Platz 2, D-97082, Germany.
Steinmeyer
R
R
Hedrich
R
R
D002462:Cell Membrane; D007887:Fabaceae / Q000502:physiology*; D007473:Ion Channels; D007700:Kinetics; D008564:Membrane Potentials; D008839:Microelectrodes; D018408:Patch-Clamp Techniques / Q000379:methods*; D019441:Plant Epidermis / Q000502:physiology; D018514:Plant Structures / Q000502:physiology; D011188:Potassium / Q000378:metabolism*
D007473:Ion Channels; D011188:Potassium