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Measurements of membrane patch capacitance using a software-based lock-in system

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Abstract

On-cell patch-clamp capacitance measurements can resolve the fusion of individual vesicles to a membrane patch and the accompanying dilation of the fusion pore. So far, these measurements have used a patch-clamp amplifier in combination with a hardware lock-in amplifier. Usually, solely the capacitance and conductance outputs of hardware lock-in amplifiers were recorded, which needed to be filtered rather heavily to suppress spectral components at the stimulus frequency. Therefore, the temporal resolution was limited, and information carried in the patch current was not utilized. In this paper, we describe an alternative and more versatile approach for measuring patch capacitance and conductance, using a digitally controlled patch-clamp amplifier. The software lock-in system showed better bandwidth and identical signal-to-noise performance needing less instrumentation. High temporal resolution measurements on patches of chromaffin cells showed that vesicle fission can be completed in only tens of microseconds. Capacitance calculation based on the patch current allows for straightforward offline phase correction. Moreover, the close inspection of direct current for the first time revealed small current changes accompanying the fusion and fission of large secretory vesicles, promising new insights into the vesicles’ membrane properties. A practical guide to high-resolution on-cell patch-clamp capacitance measurements using the software lock-in is provided.

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Acknowledgment

The experimental work and data analysis were performed by A.N. with a small contribution by C.H. We would like to thank F. Wolf for discussion and Stefan Hallermann, Manfred Heckmann and Manfred Lindau for their comments on the manuscript. We thank Jakob Neef and Andrew Woehler for participation in some recordings. We thank Margitta Köppler for excellent technical assistance and Ina Herfort for bovine chromaffin cell preparations. This work was supported by the Federal Ministry for Education and Research through the Bernstein Center for Computational Neuroscience, Goettingen, by the Deutsche Forschungsgemeinschaft (through the Center for Molecular Physiology of the Brain), by the EU (Eurohear) and by the Human Frontier Science Program Organization.

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Authors and Affiliations

  1. InnerEarLab, Department of Otolaryngology, Goettingen University Medical School, Goettingen, Germany

    Andreas Neef & Tobias Moser

  2. Bernstein Center for Computational Neuroscience, Goettingen, Germany

    Andreas Neef & Tobias Moser

  3. HEKA Elektronik Dr. Schulze GmbH, Lambrecht, Germany

    Christian Heinemann

  4. Center for Molecular Physiology of the Brain, University of Goettingen, Goettingen, Germany

    Tobias Moser

  5. Department of Otolaryngology and Bernstein Center for Computational Neuroscience, University of Goettingen, 37099, Goettingen, Germany

    Andreas Neef

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  1. Andreas Neef
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  2. Christian Heinemann
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  3. Tobias Moser
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Correspondence to Andreas Neef.

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Neef, A., Heinemann, C. & Moser, T. Measurements of membrane patch capacitance using a software-based lock-in system. Pflugers Arch - Eur J Physiol 454, 335–344 (2007). https://doi.org/10.1007/s00424-006-0191-1

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  • DOI: https://doi.org/10.1007/s00424-006-0191-1

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