Abstract
Membrane capacitance (C m) measurement, which measures and monitors plasma membrane capacitance and its changes of a cell, is one of the most important quantitative techniques and widely used in the studies of exocytosis and endocytosis. We present here a Z f-and-H sys-based C m measurement method (Z–H method) for our self-developed patch-clamp amplifier (PCA). The method adopts the complex impedance of the feedback resistor of the PCA and a transfer function, H sys, of the main and stimulus paths to generate the impedance of a patch-clamped cell, which extends Gillis’ work for C m measurements. The chief advantage of the approach is that it does not depend on idealized assumptions about the impedance supplied to the probe input by measuring the frequency responses of the various components of the PCA. H sys is generated on “frequency grids” at different standard settings of the PCA, stored and managed by a simple database, and then retrieved for calculation of cell parameters, which results in a high efficiency. An algorithm for obtaining the trans-admittance of the slow capacitance compensation (C-slow) circuitry is provided to improve the accuracy of impedance measurements when the C-slow circuitry is turned on.
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Notes
To avoid ambiguity, in H sys implementation, we use H o to symbolize the ratio of \(V_{{\text{o}}} \) to \(V_{{\text{i}}} \)
The sign of the imaginary part of H F2Out is opposite to that in Gillis’ Eq. 1 [12]. PSD detects the phase delay of the output with respect to the input, i.e., the phase difference between the reference and the measured signals [13]. What is desired here is the phase difference between the measured and reference signals; hence, the imaginary part of H F2Out should assume the minus sign
References
Heidelberger R, Heinemann C, Neher E, Matthews G (1994) Calcium dependence of the rate of exocytosis in a synaptic terminal. Nature 371:513–515
Heinemann C, Chow RH, Neher E, Zucker RS (1994) Kinetics of the secretory response in bovine chromaffin cells following flash photolysis of caged Ca2+. Biophys J 67:2546–2557
Cole KS (1968) Membranes, Ions and impulses. University of California Press, Berkeley
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
Gillis KD, Misler S (1992) Single cell assay of exocytosis from pancreatic islet β-cells. Pflugers Arch 420:121–123
Penner R, Neher E (1988) The role of calcium in stimulus-secretion coupling in excitable and non-excitable cells. J Exp Biol 139:329–345
Penner R, Neher E (1989) Stimulus-secretion coupling in mast cells. Soc Gen Physiol Ser 44:295–310
Augustine GJ, Neher E (1992) Calcium requirements for secretion in bovine chromaffin cells. J Physiol (Lond) 450:247–271
Barnett DW, Misler S (1995) Coupling of exocytosis to depolarization in rat pancreatic islet, β-cells: effects of Ca2+-, Sr2+-, and Ba2+-containing extracellular solutions. Pflugers Arch 430:593–595
Lindau M, Fernandez JM (1986) IgE-mediated degranulation of mast cells does not require opening of ion channels. Nature 319:50–153
Fernandez JM, Lindau M, Eckstein F (1987) Intracellular stimulation of mast cells with guanine nucleotides mimic antigenic stimulation. FEBS Lett 216:89–93
Gillis KD (2000) Admittance-based measurement of membrane capacitance using the EPC-9 patch-clamp amplifier. Pflugers Arch 439:655–664
Zhang H, Xiong J, Luo J, Qu AL (2008) Experimental determination of Cm measurement related hardware parameters of the patch-clamp amplifier. J Neurosci Methods. doi:10.1016/j. jneumeth.2008.08.022
Sigworth FJ (1995) Electronic design of the patch clamp. In: Sakmann B, Neher E (eds) Single-channel recording, 2nd edn. Plenum, New York, pp 95–127
Pusch M, Neher E (1988) Rats of diffusional exchange between small cell and a measuring patch pipette. Pflugers Arch 411(2):204–211
Lindau M, Neher E (1988) Patch-clamp techniques for time-resolved capacitance measurements in single cells. Pfluegers Arch 411:137–146
Blair DP, Sydenham PH (1975) Phase sensitive detection as a means to recover signals buried in noise. J Phys E Sci Instrum 8:621–627
Gillis KD (1995) Techniques for membrane capacitance measurements. In: Sakmann B, Neher E (eds) Single-channel recording, 2nd edn. Plenum, New York, pp 155–198
Donnelly DF (1994) A novel method for rapid measurement of membrane resistance, capacitance, and access resistance. Biophys J 66:873–877
Joshi C, Fernandez JM (1988) Capacitance measurements: an analysis of the phase detector technique used to study exocytosis and endocytosis. Biophys J 53:885–892
Fidler N, Fernandez JM (1989) Phase tracking: an improved phase detection technique for cell membrane capacitance measurements. Biophys J 56:1153–1162
Santos-Sacchi J (2004) Determination of cell capacitance using the exact empirical solution of ∂Y/∂Cm and its phase angle. Biophys J 87:714–727
Thompson RE, Lindau M, Webb WW (2001) Robust, high-resolution, whole cell patch-clamp capacitance measurements using square wave stimulation. Biophys J 81:937–948
Novák P, Zahradník I (2006) Q-method for high-resolution, whole-cell patch-clamp impedance measurements using square wave stimulation. Ann Biomed Eng 34:1201–1212
Acknowledgments
This work was supported by grants from National Natural Science Foundation of China (No. 60571011 and No.30327001). I wish to thank Qiuping Chen for his efforts and patience in the experiments, Zongda Wu and Zhewen Su for their tireless efforts in writing the codes.
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Zhang, H., Luo, J., Xiong, J. et al. Z f-and-H sys-based C m measurement under the whole-cell patch-clamp recording. Pflugers Arch - Eur J Physiol 457, 1423–1434 (2009). https://doi.org/10.1007/s00424-008-0614-2
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DOI: https://doi.org/10.1007/s00424-008-0614-2