Abstract
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. Particurlarly, it is equally applicable to the “slow whole-cell” configuration (access resistance in the range 100 MΩ to 1 GΩ) and to normal whole-cell measurements (access resistance typically 10 MΩ). 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.
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Almers W, Neher E (1987) Gradual and stepwise changes in the membrane capacitance of rat peritoneal mast cells. J Physiol 386:205–217
Armstrong CM, Bezanilla F (1973) Currents related to movement of the gating particles of the sodium channels. Nature 242:459–461
Clausen C, Fernandez JM (1981) A low-cost method for rapid transfer function measurements with direct application to biological impedance analysis. Pflügers Arch 390:290–295
Eisenberg RS, Mathias RT (1980) Structural analysis of electrical properties of cells and tissues. CRC Crit Rev Bioeng 4:203–232
Fernandez JM, Bezanilla F, Taylor RE (1982) Distribution and kinetics of membrane dielectric polarization. II. Frequency domain studies of gating currents. J Gen Physiol 79:41–67
Fernandez JM, Neher E, Gomperts BD (1984) Capacitance measurements reveal stepwise fusion events in degranulating mast cells. Nature 312:453–455
Fishman HM (1985) Relaxations, fluctuations and ion transfer across membranes. Prog Biophys Mol Biol 46:127–162
Fischmeister R, Ayer Jr RK, De Haan RL (1986) Some limitations of the cell-attached patch clamp technique: a two electrode analysis. Pflügers Arch 406:73–82
Gillespie JT (1979) The effect of repetitive stimulation on the passive electrical properties of the presynaptic terminal of the squid giant synapse. Proc Roy Soc Lond B 206:293–306
Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp technique for high-resolution current recording from cells and cell-free membrane patches. Pflügers Arch 391:85–100
Horowitz P, Hill W (1980) The art of electronics. Cambridge University Press, Cambridge
Jaffe LA, Hagiwara S, Kado RT (1978) The time course of cortical vesicle fusion in sea urchin eggs observed as membrane capacitance changes. Dev Biol 67:243–248
Lindau M, Fernandez JM (1986a) IgE-mediated degranulation of mast cells does not require opening of ion channels. Nature 319:150–153
Lindau M, Fernandez JM (1986b) A patch-clamp study of histamine-secreting cells. J Gen Physiol 88:349–368
Lindau M, Rüppel H (1985) On the nature of the fast light-induced charge displacement in vertebrate photoreceptors. Photobiochem Photobiophys 9:43–56
Marty A, Neher E (1983) Tight-seal whole-cell recording. In: Sakmann B, Neher E (eds) Single-channel recording. Plenum Press, New York pp 107–121
Moore LE, Christensen BN (1985) White noise analysis of cable properties of neuroblastoma cells and lamprey central neurons. J Neurophys 53:636–651
Neher E (1986) Patch clamp studies on the role of calcium in secretion. Fortschr Zool 33:275–285
Neher E (1988) The influence of intracellular calcium concentration on degranulation in dialysed mast cells from rat peritoneum. J Physiol 395 (in press)
Neher E, Marty A (1982) Discrete changes of cell membrane capacitance observed under conditions of enhanced secretion in bovine adrenal chromaffin cells. Proc Natl Acad Sci USA 79:6712–6716
Penner R, Neher E, Dreyer F (1986) Intracellularly injected tetanus toxin inhibits exocytosis in bovine adrenal chromaffin cells. Nature 324:76–78
Pusch M, Neher E (1988) Rates of diffusional exchange between small cells and a measuring patch pipette. Pflügers Arch 411:204–211
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Lindau, M., Neher, E. Patch-clamp techniques for time-resolved capacitance measurements in single cells. Pflugers Arch. 411, 137–146 (1988). https://doi.org/10.1007/BF00582306
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DOI: https://doi.org/10.1007/BF00582306