Patch-clamp techniques for time-resolved capacitance measurements in single cells.
Two methods are described for estimation of passive cell parameters such as membrane capacitance, membrane conductance and access resistance in tight-seal whole cell recording. Both methods are restricted in their application to cases where the cell under study can be approximated by a simple three-component network with linear properties over some voltage range. One method, referred to as the time domain technique, requires only standard electrophysiological equipment and a computer. Parameters are derived from an analysis of capacitive transients during square wave stimulation. It is readily adaptable to wide variations in experimental parameters. Particularly, it is equally applicable to the "slow whole-cell" configuration (access resistance in the range 100 M omega to 1 G omega) and to normal whole-cell measurements (access resistance typically 10 M omega). The other method applies a sine wave command signal to the cell and employs a lock-in amplifier to analyse the resulting current signal. Two modes of operating the lock-in amplifier are described. One mode provides an output signal directly proportional to small changes in capacitance at maximum resolution (1-10 fF). The other mode, in conjunction with a digital computer, supplies estimates of all passive cell parameters, as does the time domain technique, but with a large amount of data reduction performed by the lock-in amplifier itself. Due to the special hardware, however, this method is not as flexible as the time domain technique.
Lindau
M
M
Freie Universität Berlin, Fachbereich Physik, Abteilung Biophysik, Federal Republic of Germany.
Neher
E
E
D000818:Animals; D004553:Electric Conductivity; D004594:Electrophysiology / Q000295:instrumentation / Q000379:methods; D005089:Exocytosis; D066298:In Vitro Techniques; D007700:Kinetics; D008407:Mast Cells / Q000502:physiology*; D008564:Membrane Potentials; D013997:Time Factors