Admittance-based measurement of membrane capacitance using the EPC-9 patch-clamp amplifier

被引:90
作者
Gillis, KD
机构
[1] Univ Missouri, Dalton Cardiovasc Res Ctr, Dept Elect Engn, Columbia, MO 65211 USA
[2] Univ Missouri, Dalton Cardiovasc Res Ctr, Dept Physiol, Columbia, MO 65211 USA
[3] Max Planck Inst Biophys Chem, Dept Membrane Biophys, D-37077 Gottingen, Germany
来源
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY | 2000年 / 439卷 / 05期
关键词
endocytosis; exocytosis; lock-in amplifier; membrane capacitance; patch-clamp; phase-sensitive detector;
D O I
10.1007/s004240050990
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
A software lock-in amplifier (SLIA) was developed to allow high-time-resolution measurement of membrane capacitance as a single-cell assay of exocytosis. The unique feature of this "virtual instrument" is that it is thoroughly integrated with a computer-controlled patch-clamp amplifier (EPC-9) to allow estimation of equivalent circuit parameters based upon calibrated admittance measurements rather than just relative changes. Since the same software package ("PULSE") controls both the EPC-9 and the SLIA, instrument settings which affect admittance calculations (gain, filtering, etc.) are always "known" by the SLIA. Attenuation and phase shifts introduced within the EPC-9 by lowpass filters and other circuitry are modelled and automatically corrected by the software. In addition, changes in the measured signal introduced by whole-cell capacitance and series resistance compensation are accounted for. The noise of capacitance measurements is nearly optimal and resistive parameters can vary over a large range without inducing artifactual changes in capacitance estimates.
引用
收藏
页码:655 / 664
页数:10
相关论文
共 19 条
[1]   The exocytotic event in chromaffin cells revealed by patch amperometry [J].
Albillos, A ;
Dernick, G ;
Horstmann, H ;
Almers, W ;
deToledo, GA ;
Lindau, M .
NATURE, 1997, 389 (6650) :509-512
[2]   CALCIUM REQUIREMENTS FOR SECRETION IN BOVINE CHROMAFFIN CELLS [J].
AUGUSTINE, GJ ;
NEHER, E .
JOURNAL OF PHYSIOLOGY-LONDON, 1992, 450 :247-271
[3]   An optimized approach to membrane capacitance estimation using dual-frequency excitation [J].
Barnett, DW ;
Misler, S .
BIOPHYSICAL JOURNAL, 1997, 72 (04) :1641-1658
[4]  
COUCH LW, 1983, DIGITAL ANALOG COMMU
[5]   Influence of conductance changes on patch clamp capacitance measurements using a lock-in amplifier and limitations of the phase tracking technique [J].
Debus, K ;
Hartmann, J ;
Kilic, G ;
Lindau, M .
BIOPHYSICAL JOURNAL, 1995, 69 (06) :2808-2822
[6]   CAPACITANCE MEASUREMENTS REVEAL STEPWISE FUSION EVENTS IN DEGRANULATING MAST-CELLS [J].
FERNANDEZ, JM ;
NEHER, E ;
GOMPERTS, BD .
NATURE, 1984, 312 (5993) :453-455
[7]   PHASE TRACKING - AN IMPROVED PHASE DETECTION TECHNIQUE FOR CELL-MEMBRANE CAPACITANCE MEASUREMENTS [J].
FIDLER, N ;
FERNANDEZ, JM .
BIOPHYSICAL JOURNAL, 1989, 56 (06) :1153-1162
[8]   Protein kinase C enhances exocytosis from chromaffin cells by increasing the size of the readily releasable pool of secretory granules [J].
Gillis, KD ;
Mossner, R ;
Neher, E .
NEURON, 1996, 16 (06) :1209-1220
[9]  
Gillis Kevin D., 1995, P155
[10]   KINETICS OF THE SECRETORY RESPONSE IN BOVINE CHROMAFFIN CELLS FOLLOWING FLASH-PHOTOLYSIS OF CAGED CA2+ [J].
HEINEMANN, C ;
CHOW, RH ;
NEHER, E ;
ZUCKER, RS .
BIOPHYSICAL JOURNAL, 1994, 67 (06) :2546-2557