Sources of errors in different single-electrode voltage-clamp techniques: a computer simulation study

doi:10.1016/0165-0270(94)90176-7

Journal of Neuroscience Methods

Volume 53, Issue 2, August 1994, Pages 189-197

https://doi.org/10.1016/0165-0270(94)90176-7 Get rights and content

Abstract

The use of voltage clamp with a single electrode has been useful in estimating kinetic parameters for a number of ionic whole-cell currents. There are two main types of such a technique: discontinuous voltage clamp (dSEVC) (Brennecke and Lindemann, 1974), and continuous voltage clamp (cSEVC) (Hamill et al., 1981). We have studied, by means of computer simulations, the performance of both types of clamp on estimating activation kinetics parameters of a typical neuronal Ca²⁺ current. Deviations from the theoretical values are shown to be sensitive on both set-up and cell properties. Both types of clamp are shown to lose voltage control when either access resistance or absolute membrane conductance are increased. In contrast, changes in membrane capacitance affect differently to the estimates obtained by the two types of clamp. Cell size is also shown to affect cSEVC performance but not that of dSEVC. The nature and magnitude of errors obtained by using both types of clamp in different situations are discussed.

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Accurate measurement of junctional conductance between electrically coupled cells with dual whole-cell voltage-clamp under conditions of high series resistance
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Accurate measurement of the junctional conductance (G_j) between electrically coupled cells can provide important information about the functional properties of coupling. With the development of tight-seal, whole-cell recording, it became possible to use dual, single-electrode voltage-clamp recording from pairs of small cells to measure G_j. Experiments that require reduced perturbation of the intracellular environment can be performed with high-resistance pipettes or the perforated-patch technique, but an accompanying increase in series resistance (R_s) compromises voltage-clamp control and reduces the accuracy of G_j measurements. Here, we present a detailed analysis of methodologies available for accurate determination of steady-state G_j and related parameters under conditions of high R_s, using continuous or discontinuous single-electrode voltage-clamp (CSEVC or DSEVC) amplifiers to quantify the parameters of different equivalent electrical circuit model cells. Both types of amplifiers can provide accurate measurements of G_j, with errors less than 5% for a wide range of R_s and G_j values. However, CSEVC amplifiers need to be combined with R_s-compensation or mathematical correction for the effects of nonzero R_s and finite membrane resistance (R_m). R_s-compensation is difficult for higher values of R_s and leads to instability that can damage the recorded cells. Mathematical correction for R_s and R_m yields highly accurate results, but depends on accurate estimates of R_s throughout an experiment. DSEVC amplifiers display very accurate measurements over a larger range of R_s values than CSEVC amplifiers and have the advantage that knowledge of R_s is unnecessary, suggesting that they are preferable for long-duration experiments and/or recordings with high R_s.
Parameter-extraction of a two-compartment model for whole-cell data analysis
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Neuronal modeling of patch-clamp data is based on approximations which are valid under specific assumptions regarding cell properties and morphology. Certain cells, which show a biexponential capacitance transient decay, can be modeled with a two-compartment model. However, for parameter-extraction in such a model, approximations are required regarding the relative sizes of the various model parameters. These approximations apply to certain cell types or experimental conditions and are not valid in the general case. In this paper, we present a general method for the extraction of the parameters in a two-compartment model without assumptions regarding the relative size of the parameters. All the passive electrical parameters of the two-compartment model are derived in terms of the available experimental data. The experimental data is obtained from a DC measurement (where the command potential is a hyperpolarizing DC voltage) and an AC measurement (where the command potential is a sinusoidal stimulus on a hyperpolarized DC potential) performed on the cell under test. Computer simulations are performed with a circuit simulator, XSPICE, to observe the effects of varying the two-compartment model parameters on the capacitive transients of the current response. Our general solution for the parameter-estimation of a two-compartment model may be used to model any neuron, which has a biexponential capacitive current decay. In addition, our model avoids the need for simplifying and perhaps erroneous approximations. Our equations may be easily implemented in hardware/software compensation schemes to correct the recorded currents for any series resistance or capacitive transient errors. Our general solution reduces to the results of previous researchers under their approximations.
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A new hardware cell model for electrophysiological recording has been constructed which allows for the assessment of voltage clamp accuracy in different recording situations. Each compartment consists of a capacitor in parallel with a variable resistor and can be connected to other compartments by a variable axial resistance. The simulated membrane resistance can be changed extrinsically by a command voltage input which is optically coupled to the cell without any direct galvanic contact. Each compartment possesses a buffer amplifier which reads out the potential at the simulated membrane element, (e.g. `somatic' or `dendritic' potential). The model allows for the direct observation of typical situations and problems arising in electrophysiological experiments. We used the model to monitor deviations between the `intracellular' and the command voltage, e.g. due to series resistance errors. We also used the model to simulate synaptic currents which were generated by triangular membrane conductance changes. The results demonstrate the strong influence of synaptic location and series resistance on voltage clamp fidelity. The cell model is a new and easy-to-handle tool for the observation of voltage control under realistic experimental conditions.
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Research paperSources of errors in different single-electrode voltage-clamp techniques: a computer simulation study

Abstract

J. Neurosci. Methods

Determination of the resistance in series in the membranes of giant axons

J. Memb. Biol.

Theory of membrane voltage clamp with discontinuous feedback through a pulsed current clamp

Rev. Sci. Instr.

Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches

Pflügers Arch.

A quantitative description of membrane current and its application to conduction and excitation in nerve

J. Physiol.

Sodium currents in dissociated bull-frog sympathetic neurones

J. Physiol.

Calcium currents in bullfrog sympathetic neurons. I. Activation kinetics and pharmacology

J. Gen. Physiol.

Research paper
Sources of errors in different single-electrode voltage-clamp techniques: a computer simulation study