Patch Clamp

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Niels Fertig - One of the best experts on this subject based on the ideXlab platform.

  • Characterization of iPS Derived Cardiomyocytes in Voltage Clamp and Current Clamp by Automated Patch Clamp
    Biophysical Journal, 2017
    Co-Authors: Andrea Brüggemann, Claudia Haarmann, Michael George, Markus Rapedius, Tom A. Goetze, Ilka Rinke, Niels Fertig
    Abstract:

    In recent years, human stem cell-derived cardiomyocytes have proven to recapitulate key features of human cardiac electrophysiology in vitro. Furthermore, it has become apparent that the intact ensemble of cardiac ion channels is necessary to determine proarrhythmic effects reliably. Hence, due to their increasing availability, stem cell-derived cardiomyocytes have become the preferred choice of cardiac cells.This poster summarizes the promises and challenges of combining iPS derived cardiac myocytes with automated Patch Clamp. Features like high throughput, temperature control, easy internal solution exchange and full automation make planar Patch Clamp a desired method for characterizing iPS derived cardiomyocytes.One of the biggest challenges of planar Patch Clamp in this context is the fact that individual cells cannot be chosen, but cells will be selected randomly. In addition cells have to be harvested before the application to the Patch Clamp chip and cannot be Patched as adherent cells.With this poster, we show our progress on these challenges. Two automated Patch Clamp platforms, the Patchliner, as a medium throughput, and the SyncroPatch 384PE, as a high throughput device, were used for this study.Pharmacological measurements in voltage Clamp as well as current Clamp will be shown also under physiological temperatures and perforated Patch.

  • comprehensive cardiac safety screening pharmacology of stem cell derived cardiomyocytes using high throughput automated Patch Clamp
    Biophysical Journal, 2016
    Co-Authors: Nadine Becker, Claudia Haarmann, Michael George, Markus Rapedius, Ilka Rinke, Andrea Brueggemann, Timo Stengel, Sonja Stolzlefeix, Tom Gotze, Niels Fertig
    Abstract:

    Drug discovery is challenged by poor predictability of proarrhythmic effects of present safety screening approaches. The Comprehensive in Vitro Proarrhythmia Assay (CiPA), an FDA-directed initiative, is specifically focused on improved preclinical assessment of torsade de pointes (TdP) risk, striving to optimize the drug discovery process.In recent years, human stem cell-derived cardiomyocytes have proven to recapitulate key features of human cardiac electrophysiology in vitro. Furthermore, it has become apparent that the intact ensemble of cardiac ion channels is necessary to determine proarrhythmic effects reliably. Hence, due to their relatively easy availability, stem cell-derived cardiomyocytes have become the preferred choice of cardiac cells.We present chip-based approaches allowing parallel Patch Clamp recordings without compromising data quality or technical sophistication. Culture and harvesting protocols have been optimized on cardiomyocytes for success rate but also the number of cells used.We show Patch Clamp data from two automated Patch Clamp platforms (medium throughput - Patchliner and high throughput - SyncroPatch 384/768PE). The Patchliner and the SyncroPatch 384/768PE both boast temperature control to enable recordings at physiological temperature - an important feature since drug efficacies may vary with temperature. Here, we present pharmacological data on voltage-dependent channels of the CiPA panel at different temperatures.The effect of drugs on action potentials is important for assessing the interaction of the cardiac ion channel ensemble. Both Patch Clamp platforms have amplifiers capable of current Clamp. Compound effects on action potentials will also be presented.Patchliner and SyncroPatch 384/768PE are both planar Patch-Clamp platforms combining all the necessary features for successful drug discovery (well-suited for stem cell-derived cardiomyocytes, temperature control, and current Clamp mode).

  • automated Patch Clamp analysis of nachα7 and nav1 7 channels
    Current protocols in pharmacology, 2014
    Co-Authors: Alison Obergrussberger, Claudia Haarmann, Michael George, Ilka Rinke, David Guinot, Sonja Stoelzlefeix, Nadine Becker, Andrea Brueggemann, Niels Fertig
    Abstract:

    Automated Patch Clamp devices are now commonly used for studying ion channels. A useful modification of this approach is the replacement of the glass pipet with a thin planar glass layer with a small hole in the middle. Planar Patch Clamp devices, such as the three described in this unit, are overtaking glass pipets in popularity because they increase throughput, are easier to use, provide for the acquisition of high-quality and information-rich data, and allow for rapid perfusion and temperature control. Covered in this unit are two challenging targets in drug discovery: voltage-gated sodium subtype 1.7 (NaV1.7) and nicotinic acetylcholine α7 receptors (nAChα7R). Provided herein are protocols for recording activation and inactivation kinetics of NaV1.7, and activation and allosteric modulation of nAChα7R. Curr. Protoc. Pharmacol. 65:11.13.1-11.13.48. © 2014 by John Wiley & Sons, Inc. Keywords: automated Patch Clamp; NaV1.7; nicotinic acetylcholine receptors

  • hts techniques for Patch Clamp based ion channel screening advances and economy
    Expert Opinion on Drug Discovery, 2012
    Co-Authors: Cecilia Farre, Niels Fertig
    Abstract:

    Introduction: Ten years ago, the first publication appeared showing Patch Clamp recordings performed on a planar glass chip instead of using a conventional Patch Clamp pipette. “Going planar” proved to revolutionize ion channel drug screening as we know it, by allowing high quality measurements of ion channels and their effectors at a higher throughput and at the same time de-skilling the highly laborious technique. Over the years, platforms evolved in response to user requirements regarding experimental features, data handling plus storage, and suitable target diversity. Areas covered: This article gives a snapshot image of Patch Clamp-based ion channel screening with focus on platforms developed to meet requirements of high-throughput screening environments. The commercially available platforms are described, along with their benefits and drawbacks in ion channel drug screening. Expert opinion: Automated Patch Clamp (APC) platforms allow faster investigation of a larger number of ion channel active comp...

  • automated Patch Clamp with current Clamp action potential recordings from stem cell derived cardiomyocytes
    Biophysical Journal, 2011
    Co-Authors: Sonja Stoelzle, Andrea Brüggemann, Alison Haythornthwaite, Claudia Haarmann, Cecilia Farre, Rodolfo J Haedo, Michael George, Niels Fertig
    Abstract:

    Automated planar Patch Clamp devices, with their higher throughput and high data information content, are finding their place in the market for making high quality electrophysiological measurements.Ease of use and higher data throughput make these devices ideal tools for ion channel screening and safety testing. The ability to record from stem cell derived cardiomyocytes in both voltage and current Clamp modes on an automated Patch Clamp platform is an important advancement in ion channel screening and safety testing. Using a planar Patch Clamp workstation, currents mediated by K+, Na+ and Ca2+ channels could be recorded from stem cell derived cardiomyocytes in the voltage Clamp mode. In the current Clamp mode, action potentials could be recorded for the first time on a planar Patch Clamp device. The built-in temperature control allowed for pharmacology on action potentials (see figure) both at room temperature and 35°.The use of stem cell derived cardiomyocytes in safety testing is becoming increasingly important. The ability to test compounds on ion channels in both voltage and current Clamp modes, as well as at different temperatures, may be crucial for future safety testing.View Large Image | View Hi-Res Image | Download PowerPoint Slide

Michael George - One of the best experts on this subject based on the ideXlab platform.

  • Characterization of iPS Derived Cardiomyocytes in Voltage Clamp and Current Clamp by Automated Patch Clamp
    Biophysical Journal, 2017
    Co-Authors: Andrea Brüggemann, Claudia Haarmann, Michael George, Markus Rapedius, Tom A. Goetze, Ilka Rinke, Niels Fertig
    Abstract:

    In recent years, human stem cell-derived cardiomyocytes have proven to recapitulate key features of human cardiac electrophysiology in vitro. Furthermore, it has become apparent that the intact ensemble of cardiac ion channels is necessary to determine proarrhythmic effects reliably. Hence, due to their increasing availability, stem cell-derived cardiomyocytes have become the preferred choice of cardiac cells.This poster summarizes the promises and challenges of combining iPS derived cardiac myocytes with automated Patch Clamp. Features like high throughput, temperature control, easy internal solution exchange and full automation make planar Patch Clamp a desired method for characterizing iPS derived cardiomyocytes.One of the biggest challenges of planar Patch Clamp in this context is the fact that individual cells cannot be chosen, but cells will be selected randomly. In addition cells have to be harvested before the application to the Patch Clamp chip and cannot be Patched as adherent cells.With this poster, we show our progress on these challenges. Two automated Patch Clamp platforms, the Patchliner, as a medium throughput, and the SyncroPatch 384PE, as a high throughput device, were used for this study.Pharmacological measurements in voltage Clamp as well as current Clamp will be shown also under physiological temperatures and perforated Patch.

  • comprehensive cardiac safety screening pharmacology of stem cell derived cardiomyocytes using high throughput automated Patch Clamp
    Biophysical Journal, 2016
    Co-Authors: Nadine Becker, Claudia Haarmann, Michael George, Markus Rapedius, Ilka Rinke, Andrea Brueggemann, Timo Stengel, Sonja Stolzlefeix, Tom Gotze, Niels Fertig
    Abstract:

    Drug discovery is challenged by poor predictability of proarrhythmic effects of present safety screening approaches. The Comprehensive in Vitro Proarrhythmia Assay (CiPA), an FDA-directed initiative, is specifically focused on improved preclinical assessment of torsade de pointes (TdP) risk, striving to optimize the drug discovery process.In recent years, human stem cell-derived cardiomyocytes have proven to recapitulate key features of human cardiac electrophysiology in vitro. Furthermore, it has become apparent that the intact ensemble of cardiac ion channels is necessary to determine proarrhythmic effects reliably. Hence, due to their relatively easy availability, stem cell-derived cardiomyocytes have become the preferred choice of cardiac cells.We present chip-based approaches allowing parallel Patch Clamp recordings without compromising data quality or technical sophistication. Culture and harvesting protocols have been optimized on cardiomyocytes for success rate but also the number of cells used.We show Patch Clamp data from two automated Patch Clamp platforms (medium throughput - Patchliner and high throughput - SyncroPatch 384/768PE). The Patchliner and the SyncroPatch 384/768PE both boast temperature control to enable recordings at physiological temperature - an important feature since drug efficacies may vary with temperature. Here, we present pharmacological data on voltage-dependent channels of the CiPA panel at different temperatures.The effect of drugs on action potentials is important for assessing the interaction of the cardiac ion channel ensemble. Both Patch Clamp platforms have amplifiers capable of current Clamp. Compound effects on action potentials will also be presented.Patchliner and SyncroPatch 384/768PE are both planar Patch-Clamp platforms combining all the necessary features for successful drug discovery (well-suited for stem cell-derived cardiomyocytes, temperature control, and current Clamp mode).

  • automated Patch Clamp analysis of nachα7 and nav1 7 channels
    Current protocols in pharmacology, 2014
    Co-Authors: Alison Obergrussberger, Claudia Haarmann, Michael George, Ilka Rinke, David Guinot, Sonja Stoelzlefeix, Nadine Becker, Andrea Brueggemann, Niels Fertig
    Abstract:

    Automated Patch Clamp devices are now commonly used for studying ion channels. A useful modification of this approach is the replacement of the glass pipet with a thin planar glass layer with a small hole in the middle. Planar Patch Clamp devices, such as the three described in this unit, are overtaking glass pipets in popularity because they increase throughput, are easier to use, provide for the acquisition of high-quality and information-rich data, and allow for rapid perfusion and temperature control. Covered in this unit are two challenging targets in drug discovery: voltage-gated sodium subtype 1.7 (NaV1.7) and nicotinic acetylcholine α7 receptors (nAChα7R). Provided herein are protocols for recording activation and inactivation kinetics of NaV1.7, and activation and allosteric modulation of nAChα7R. Curr. Protoc. Pharmacol. 65:11.13.1-11.13.48. © 2014 by John Wiley & Sons, Inc. Keywords: automated Patch Clamp; NaV1.7; nicotinic acetylcholine receptors

  • automated Patch Clamp with current Clamp action potential recordings from stem cell derived cardiomyocytes
    Biophysical Journal, 2011
    Co-Authors: Sonja Stoelzle, Andrea Brüggemann, Alison Haythornthwaite, Claudia Haarmann, Cecilia Farre, Rodolfo J Haedo, Michael George, Niels Fertig
    Abstract:

    Automated planar Patch Clamp devices, with their higher throughput and high data information content, are finding their place in the market for making high quality electrophysiological measurements.Ease of use and higher data throughput make these devices ideal tools for ion channel screening and safety testing. The ability to record from stem cell derived cardiomyocytes in both voltage and current Clamp modes on an automated Patch Clamp platform is an important advancement in ion channel screening and safety testing. Using a planar Patch Clamp workstation, currents mediated by K+, Na+ and Ca2+ channels could be recorded from stem cell derived cardiomyocytes in the voltage Clamp mode. In the current Clamp mode, action potentials could be recorded for the first time on a planar Patch Clamp device. The built-in temperature control allowed for pharmacology on action potentials (see figure) both at room temperature and 35°.The use of stem cell derived cardiomyocytes in safety testing is becoming increasingly important. The ability to test compounds on ion channels in both voltage and current Clamp modes, as well as at different temperatures, may be crucial for future safety testing.View Large Image | View Hi-Res Image | Download PowerPoint Slide

  • Excitement Over Automated Patch Clamp: Action Potentials from Cardiac Myocytes
    Biophysical Journal, 2010
    Co-Authors: Sonja Stoelzle, Andrea Brüggemann, Alison Haythornthwaite, Claudia Haarmann, Cecilia Farre, Michael George, David Guinot, Ralf Kettenhofen, Niels Fertig
    Abstract:

    The use of cardiac myocytes is becoming increasingly important for drug safety testing. Unique features of certain planar Patch Clamp workstations, coupled with ease-of-use and higher data throughput, make these devices ideal tools for ion channel screening and safety testing. Using stem cell derived cardiac myocytes, recordings could be made not only in the voltage-Clamp mode but also in the current-Clamp mode on a planar Patch Clamp workstation. This demonstrates for the first time parallel current-Clamp recordings on a planar Patch Clamp workstation. Ion channels important in drug discovery, such as hERG and voltage-gated Na+, Ca2+ and K+ channels in the voltage-Clamp mode from stem cell derived cardiac myocytes will be shown. In addition, action potential recordings in the current-Clamp mode at 35°C, and modulation of the action potentials by hERG active compounds, will also be shown.View Large Image | View Hi-Res Image | Download PowerPoint Slide

Andrea Brüggemann - One of the best experts on this subject based on the ideXlab platform.

  • Characterization of iPS Derived Cardiomyocytes in Voltage Clamp and Current Clamp by Automated Patch Clamp
    Biophysical Journal, 2017
    Co-Authors: Andrea Brüggemann, Claudia Haarmann, Michael George, Markus Rapedius, Tom A. Goetze, Ilka Rinke, Niels Fertig
    Abstract:

    In recent years, human stem cell-derived cardiomyocytes have proven to recapitulate key features of human cardiac electrophysiology in vitro. Furthermore, it has become apparent that the intact ensemble of cardiac ion channels is necessary to determine proarrhythmic effects reliably. Hence, due to their increasing availability, stem cell-derived cardiomyocytes have become the preferred choice of cardiac cells.This poster summarizes the promises and challenges of combining iPS derived cardiac myocytes with automated Patch Clamp. Features like high throughput, temperature control, easy internal solution exchange and full automation make planar Patch Clamp a desired method for characterizing iPS derived cardiomyocytes.One of the biggest challenges of planar Patch Clamp in this context is the fact that individual cells cannot be chosen, but cells will be selected randomly. In addition cells have to be harvested before the application to the Patch Clamp chip and cannot be Patched as adherent cells.With this poster, we show our progress on these challenges. Two automated Patch Clamp platforms, the Patchliner, as a medium throughput, and the SyncroPatch 384PE, as a high throughput device, were used for this study.Pharmacological measurements in voltage Clamp as well as current Clamp will be shown also under physiological temperatures and perforated Patch.

  • Studying mechanosensitive ion channels with an automated Patch Clamp
    European Biophysics Journal, 2014
    Co-Authors: Maria Barthmes, Mac Donald F Jose, Jan Peter Birkner, Andrea Brüggemann, Christian Wahl-schott, Armaǧan Koçer
    Abstract:

    Patch Clamp electrophysiology is the main technique to study mechanosensitive ion channels (MSCs), however, conventional Patch Clamping is laborious and success and output depends on the skills of the operator. Even though automated Patch systems solve these problems for other ion channels, they could not be applied to MSCs. Here, we report on activation and single channel analysis of a bacterial mechanosensitive ion channel using an automated Patch Clamp system. With the automated system, we could Patch not only giant unilamellar liposomes but also giant Escherichia coli (E. coli) spheroplasts. The tension sensitivity and channel kinetics data obtained in the automated system were in good agreement with that obtained from the conventional Patch Clamp. The findings will pave the way to high throughput fundamental and drug screening studies on mechanosensitive ion channels.

  • automated Patch Clamp with current Clamp action potential recordings from stem cell derived cardiomyocytes
    Biophysical Journal, 2011
    Co-Authors: Sonja Stoelzle, Andrea Brüggemann, Alison Haythornthwaite, Claudia Haarmann, Cecilia Farre, Rodolfo J Haedo, Michael George, Niels Fertig
    Abstract:

    Automated planar Patch Clamp devices, with their higher throughput and high data information content, are finding their place in the market for making high quality electrophysiological measurements.Ease of use and higher data throughput make these devices ideal tools for ion channel screening and safety testing. The ability to record from stem cell derived cardiomyocytes in both voltage and current Clamp modes on an automated Patch Clamp platform is an important advancement in ion channel screening and safety testing. Using a planar Patch Clamp workstation, currents mediated by K+, Na+ and Ca2+ channels could be recorded from stem cell derived cardiomyocytes in the voltage Clamp mode. In the current Clamp mode, action potentials could be recorded for the first time on a planar Patch Clamp device. The built-in temperature control allowed for pharmacology on action potentials (see figure) both at room temperature and 35°.The use of stem cell derived cardiomyocytes in safety testing is becoming increasingly important. The ability to test compounds on ion channels in both voltage and current Clamp modes, as well as at different temperatures, may be crucial for future safety testing.View Large Image | View Hi-Res Image | Download PowerPoint Slide

  • Excitement Over Automated Patch Clamp: Action Potentials from Cardiac Myocytes
    Biophysical Journal, 2010
    Co-Authors: Sonja Stoelzle, Andrea Brüggemann, Alison Haythornthwaite, Claudia Haarmann, Cecilia Farre, Michael George, David Guinot, Ralf Kettenhofen, Niels Fertig
    Abstract:

    The use of cardiac myocytes is becoming increasingly important for drug safety testing. Unique features of certain planar Patch Clamp workstations, coupled with ease-of-use and higher data throughput, make these devices ideal tools for ion channel screening and safety testing. Using stem cell derived cardiac myocytes, recordings could be made not only in the voltage-Clamp mode but also in the current-Clamp mode on a planar Patch Clamp workstation. This demonstrates for the first time parallel current-Clamp recordings on a planar Patch Clamp workstation. Ion channels important in drug discovery, such as hERG and voltage-gated Na+, Ca2+ and K+ channels in the voltage-Clamp mode from stem cell derived cardiac myocytes will be shown. In addition, action potential recordings in the current-Clamp mode at 35°C, and modulation of the action potentials by hERG active compounds, will also be shown.View Large Image | View Hi-Res Image | Download PowerPoint Slide

  • ion channel screening automated Patch Clamp on the rise
    Drug Discovery Today: Technologies, 2008
    Co-Authors: Cecilia Farre, Andrea Brüggemann, Michael George, Niels Fertig
    Abstract:

    Ion channel proteins are of major importance for the human physiology and thus highly attractive molecular drug targets. Large-scale ion channel screening of wanted and unwanted drug effects is required, but has been limited by the lack of adequate screening technology, because available methods put a trade-off between high-throughput and high-information content. The advent of automated Patch Clamp platforms has revolutionized ion channel screening, enabling investigations from a more functional perspective at a much higher throughput. The current status of automated Patch Clamp platforms, their strengths and drawbacks as well as future developments are reviewed.

Claudia Haarmann - One of the best experts on this subject based on the ideXlab platform.

  • Characterization of iPS Derived Cardiomyocytes in Voltage Clamp and Current Clamp by Automated Patch Clamp
    Biophysical Journal, 2017
    Co-Authors: Andrea Brüggemann, Claudia Haarmann, Michael George, Markus Rapedius, Tom A. Goetze, Ilka Rinke, Niels Fertig
    Abstract:

    In recent years, human stem cell-derived cardiomyocytes have proven to recapitulate key features of human cardiac electrophysiology in vitro. Furthermore, it has become apparent that the intact ensemble of cardiac ion channels is necessary to determine proarrhythmic effects reliably. Hence, due to their increasing availability, stem cell-derived cardiomyocytes have become the preferred choice of cardiac cells.This poster summarizes the promises and challenges of combining iPS derived cardiac myocytes with automated Patch Clamp. Features like high throughput, temperature control, easy internal solution exchange and full automation make planar Patch Clamp a desired method for characterizing iPS derived cardiomyocytes.One of the biggest challenges of planar Patch Clamp in this context is the fact that individual cells cannot be chosen, but cells will be selected randomly. In addition cells have to be harvested before the application to the Patch Clamp chip and cannot be Patched as adherent cells.With this poster, we show our progress on these challenges. Two automated Patch Clamp platforms, the Patchliner, as a medium throughput, and the SyncroPatch 384PE, as a high throughput device, were used for this study.Pharmacological measurements in voltage Clamp as well as current Clamp will be shown also under physiological temperatures and perforated Patch.

  • comprehensive cardiac safety screening pharmacology of stem cell derived cardiomyocytes using high throughput automated Patch Clamp
    Biophysical Journal, 2016
    Co-Authors: Nadine Becker, Claudia Haarmann, Michael George, Markus Rapedius, Ilka Rinke, Andrea Brueggemann, Timo Stengel, Sonja Stolzlefeix, Tom Gotze, Niels Fertig
    Abstract:

    Drug discovery is challenged by poor predictability of proarrhythmic effects of present safety screening approaches. The Comprehensive in Vitro Proarrhythmia Assay (CiPA), an FDA-directed initiative, is specifically focused on improved preclinical assessment of torsade de pointes (TdP) risk, striving to optimize the drug discovery process.In recent years, human stem cell-derived cardiomyocytes have proven to recapitulate key features of human cardiac electrophysiology in vitro. Furthermore, it has become apparent that the intact ensemble of cardiac ion channels is necessary to determine proarrhythmic effects reliably. Hence, due to their relatively easy availability, stem cell-derived cardiomyocytes have become the preferred choice of cardiac cells.We present chip-based approaches allowing parallel Patch Clamp recordings without compromising data quality or technical sophistication. Culture and harvesting protocols have been optimized on cardiomyocytes for success rate but also the number of cells used.We show Patch Clamp data from two automated Patch Clamp platforms (medium throughput - Patchliner and high throughput - SyncroPatch 384/768PE). The Patchliner and the SyncroPatch 384/768PE both boast temperature control to enable recordings at physiological temperature - an important feature since drug efficacies may vary with temperature. Here, we present pharmacological data on voltage-dependent channels of the CiPA panel at different temperatures.The effect of drugs on action potentials is important for assessing the interaction of the cardiac ion channel ensemble. Both Patch Clamp platforms have amplifiers capable of current Clamp. Compound effects on action potentials will also be presented.Patchliner and SyncroPatch 384/768PE are both planar Patch-Clamp platforms combining all the necessary features for successful drug discovery (well-suited for stem cell-derived cardiomyocytes, temperature control, and current Clamp mode).

  • automated Patch Clamp analysis of nachα7 and nav1 7 channels
    Current protocols in pharmacology, 2014
    Co-Authors: Alison Obergrussberger, Claudia Haarmann, Michael George, Ilka Rinke, David Guinot, Sonja Stoelzlefeix, Nadine Becker, Andrea Brueggemann, Niels Fertig
    Abstract:

    Automated Patch Clamp devices are now commonly used for studying ion channels. A useful modification of this approach is the replacement of the glass pipet with a thin planar glass layer with a small hole in the middle. Planar Patch Clamp devices, such as the three described in this unit, are overtaking glass pipets in popularity because they increase throughput, are easier to use, provide for the acquisition of high-quality and information-rich data, and allow for rapid perfusion and temperature control. Covered in this unit are two challenging targets in drug discovery: voltage-gated sodium subtype 1.7 (NaV1.7) and nicotinic acetylcholine α7 receptors (nAChα7R). Provided herein are protocols for recording activation and inactivation kinetics of NaV1.7, and activation and allosteric modulation of nAChα7R. Curr. Protoc. Pharmacol. 65:11.13.1-11.13.48. © 2014 by John Wiley & Sons, Inc. Keywords: automated Patch Clamp; NaV1.7; nicotinic acetylcholine receptors

  • automated Patch Clamp with current Clamp action potential recordings from stem cell derived cardiomyocytes
    Biophysical Journal, 2011
    Co-Authors: Sonja Stoelzle, Andrea Brüggemann, Alison Haythornthwaite, Claudia Haarmann, Cecilia Farre, Rodolfo J Haedo, Michael George, Niels Fertig
    Abstract:

    Automated planar Patch Clamp devices, with their higher throughput and high data information content, are finding their place in the market for making high quality electrophysiological measurements.Ease of use and higher data throughput make these devices ideal tools for ion channel screening and safety testing. The ability to record from stem cell derived cardiomyocytes in both voltage and current Clamp modes on an automated Patch Clamp platform is an important advancement in ion channel screening and safety testing. Using a planar Patch Clamp workstation, currents mediated by K+, Na+ and Ca2+ channels could be recorded from stem cell derived cardiomyocytes in the voltage Clamp mode. In the current Clamp mode, action potentials could be recorded for the first time on a planar Patch Clamp device. The built-in temperature control allowed for pharmacology on action potentials (see figure) both at room temperature and 35°.The use of stem cell derived cardiomyocytes in safety testing is becoming increasingly important. The ability to test compounds on ion channels in both voltage and current Clamp modes, as well as at different temperatures, may be crucial for future safety testing.View Large Image | View Hi-Res Image | Download PowerPoint Slide

  • Excitement Over Automated Patch Clamp: Action Potentials from Cardiac Myocytes
    Biophysical Journal, 2010
    Co-Authors: Sonja Stoelzle, Andrea Brüggemann, Alison Haythornthwaite, Claudia Haarmann, Cecilia Farre, Michael George, David Guinot, Ralf Kettenhofen, Niels Fertig
    Abstract:

    The use of cardiac myocytes is becoming increasingly important for drug safety testing. Unique features of certain planar Patch Clamp workstations, coupled with ease-of-use and higher data throughput, make these devices ideal tools for ion channel screening and safety testing. Using stem cell derived cardiac myocytes, recordings could be made not only in the voltage-Clamp mode but also in the current-Clamp mode on a planar Patch Clamp workstation. This demonstrates for the first time parallel current-Clamp recordings on a planar Patch Clamp workstation. Ion channels important in drug discovery, such as hERG and voltage-gated Na+, Ca2+ and K+ channels in the voltage-Clamp mode from stem cell derived cardiac myocytes will be shown. In addition, action potential recordings in the current-Clamp mode at 35°C, and modulation of the action potentials by hERG active compounds, will also be shown.View Large Image | View Hi-Res Image | Download PowerPoint Slide

Fred J. Sigworth - One of the best experts on this subject based on the ideXlab platform.

  • Patch Clamp amplifiers on a chip
    Journal of Neuroscience Methods, 2010
    Co-Authors: P Weerakoon, Eugenio Culurciello, Youshan Yang, Joseph Santossacchi, Peter J Kindlmann, Fred J. Sigworth
    Abstract:

    We present the first, fully integrated, two-channel implementation of a Patch-Clamp measurement system. With this "PatchChip" two simultaneous whole-cell recordings can be obtained with rms noise of 8pA in a 10kHz bandwidth. The capacitance and series-resistance of the electrode can be compensated up to 10pF and 100MΩ respectively under computer control. Recordings of hERG and Na(v) 1.7 currents demonstrate the system's capabilities, which are on par with large, commercial Patch-Clamp instrumentation. By reducing Patch-Clamp amplifiers to a millimeter size micro-chip, this work paves the way to the realization of massively parallel, high-throughput Patch-Clamp systems for drug screening and ion-channel research. The PatchChip is implemented in a 0.5μm silicon-on-sapphire process; its size is 3×3mm(2) and the power consumption is 5mW per channel with a 3.3V power supply.

  • an integrated Patch Clamp potentiostat with electrode compensation
    IEEE Transactions on Biomedical Circuits and Systems, 2009
    Co-Authors: P Weerakoon, Eugenio Culurciello, Kathryn G. Klemic, Fred J. Sigworth
    Abstract:

    We present the first fully integrated implementation of a Patch-Clamp measurement system with series-access resistance and parasitic capacitive compensation capability. The system was implemented in a 0.5- mum silicon-on-sapphire process and is capable of recording cell membrane currents up to plusmn20 nA, with an rms noise of 5 pA at 10-kHz bandwidth. The system can compensate for the capacitance and resistance of the electrode, up to 20 pF and up to 70% of the series-access resistance, respectively. The die size is 1150 by 700 mum. The power consumption is 300 muW at 3.3 V. The integrated Patch-Clamp system will be used to fabricate high-throughput planar Patch-Clamp systems.

  • An integrated Patch-Clamp amplifier in silicon-on-sapphire CMOS
    2006 IEEE International Symposium on Circuits and Systems, 2006
    Co-Authors: Farah Laiwalla, Kathryn G. Klemic, Fred J. Sigworth, Eugenio Culurciello
    Abstract:

    We fabricated an integrated Patch-Clamp amplifier capable of recording from pico- to tens of micro-amperes of current. The high-dynamic range of seven decades and the pico-ampere sensitivity of the instrument was designed for whole-cell Patch-Clamp recordings. The prototype was fabricated on a 0.5mum silicon-on-sapphire process. The device employs an integrating headstage with a frequency-modulated output pulse ranging from 3Hz to 10MHz. A digital interface produces a 16bit output conversion of the input currents. We report on electrical measurements from the fabricated device, and measurements conducted on cells in a typical Patch-Clamp experiment

  • microfluidic system for planar Patch Clamp electrode arrays
    Nano Letters, 2006
    Co-Authors: Kathryn G. Klemic, Mark A Reed, Fred J. Sigworth
    Abstract:

    We present a microfluidic system integrated with disposable cell interface partitions for simultaneous Patch Clamp recordings. Glass-supported poly(dimethylsiloxane) (PDMS) partitions, having a 2 microm air-blown aperture, were reversibly sealed to a microfluidic system including PDMS channels with isolation valves and microfabricated Ag/AgCl electrodes. Gigaseal recordings from RBL-1 cells were obtained with a 24% success rate. Simultaneous whole cell recordings from valve-isolated electrodes were obtained.

  • An Integrated Patch-Clamp Amplifier in Silicon-on-Sapphire CMOS
    IEEE Transactions on Circuits and Systems I: Regular Papers, 2006
    Co-Authors: Farah Laiwalla, Kathryn G. Klemic, Fred J. Sigworth, Eugenio Culurciello
    Abstract:

    We designed and tested an integrated Patch-Clamp amplifier capable of recording from pico to tens of microamperes of current. The high-dynamic range of seven decades and the picoampere sensitivity of the instrument was targeted to whole-cell Patch-Clamp recordings. The prototype was fabricated on a 0.5-mum silicon-on-sapphire process. The device employs an integrating headstage with a pulse frequency modulated output, ranging from 3 Hz to 10 MHz. A digital interface produces a 16-bit output conversion of the input currents. We report on electronic characterization of the fabricated device, dynamic performance, and examples of measurements on biological cells for Patch-Clamp applications. The device will be used in an advanced planar high-throughput Patch-Clamp screening system for testing medicines