Patch-clamp recording is a widely used technique for studying ion channel functions and cellular responses. During patch-clamp experiments, the fast capacitance (CFast) transient is a detrimental artifact and must be eliminated with CFast compensation circuit. When applying the conventional computer-aided CFast compensation procedure, the patch-clamp amplifier faces the risk of saturation due to large square-wave responses, and compensation errors may be caused due to inherent disadvantages of iterative algorithm. Here, we introduce a novel non-iterative automatic CFast compensation method, namely K-method, based on kernel estimation with white noise excitation. The kernel was estimated with cross-correlation technique and captured dynamic properties of the CFast and its related hardware. To achieve the optimal settings of the CFast compensation, the kernel of CFast was fitted to two calibrated kernels indicating two distinct types of compensation effect: the “instantaneous” and “delay” effect. The fitted coefficients were used to adjust compensation circuit. White noise excitation significantly reduced the possibility of saturation, and the K-method suffered from no typical disadvantage of iterative method. We performed compensation experiments on a model circuit and HEK293 cells. The results demonstrated a good accuracy of the K-method and the membrane capacitance measurement could benefit from it.
This is a preview of subscription content, log in to check access.
Buy single article
Instant unlimited access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Hamill, O. P., A. Marty, E. Neher, B. Sakmann, and F. J. Sigworth. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 391:85–100, 1981.
Marmarelis, V. Z. Nonlinear Dynamic Modeling of Physiological Systems. New Jersey: Wiley, 541 pp, 2004.
Marty, A., and E. Neher. Tight-seal whole-cell recording. In: Single-Channel Recording, edited by B. Sakmann, and E. Neher. New York: Plenum Press, 1995, pp. 31–52.
Press, W., S. Teukolsky, W. T. Vetterling, and B. Flannery. Numerical Recipes in C: the Art of Scientific Computing. New York: Cambridge University Press, 1235 pp., 2007.
Sigworth, F. J. Design of the EPC-9, a computer-controlled patch-clamp amplifier. 1. Hardware. J. Neurosci. Methods 56:195–202, 1995.
Sigworth, F. J., H. Affolter, and E. Neher. Design of the EPC-9, a computer-controlled patch-clamp amplifier. 2. Software. J. Neurosci. Methods 56:203–215, 1995.
Xiong, J., G. Hu, and A. Qu. Automatic calibration of frequency compensation system in computer-controlled patch-clamp amplifier. J. Comp. Sci. 3:765–772, 2007.
Ye, Y., G. Hu, and A. Qu. The improvement of C-Fast transient cancellation in automatic patch-clamp amplifier. Shanghai J. Biomed. Eng. 27:98–101, 2006.
Zhang, H., J. Luo, J. Xiong, X. G. Lin, Z. X. Wu, and A. Qu. Zf-and-Hsys-based Cm measurement under the whole-cell patch-clamp recording. Pflugers Arch. 457:1423–1434, 2009.
Zhang, H., A. Qu, J. Luo, and J. Luo. Error analysis of Cm measurement under the whole-cell patch-clamp recording. J. Neurosci. Methods 185:307–314, 2010.
Zhang, H., J. Xiong, J. Luo, and A. Qu. Experimental determination of Cm measurement related hardware parameters of the patch-clamp amplifier. J. Neurosci. Methods. 176:246–253, 2009.
We thank Haowen Liu for his efforts and patience in the experiments. This work was supported by grants from National Nature Science Foundation of China (No. 30327001).
Conflict of interest
The authors have declared that no conflict of interest exists.
Associate Editor Edward Guo oversaw the review of this article.
About this article
Cite this article
Luo, J., Hou, P., Ding, J. et al. Kernel Based Non-Iterative Automatic Fast Capacitance Compensation in Patch-Clamp Experiments. Cel. Mol. Bioeng. 5, 440–449 (2012). https://doi.org/10.1007/s12195-012-0241-z
- White noise excitation
- Cross-correlation technique
- Membrane capacitance measurement