Articles de revues : « Automated patch clamp »

1

Shieh, Char-Chang. « Automated high-throughput patch-clamp techniques ». Drug Discovery Today 9, n^o 13 (juillet 2004) : 551–52. http://dx.doi.org/10.1016/s1359-6446(04)03113-7.

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Annecchino, Luca A., et Simon R. Schultz. « Progress in automating patch clamp cellular physiology ». Brain and Neuroscience Advances 2 (1 janvier 2018) : 239821281877656. http://dx.doi.org/10.1177/2398212818776561.

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Patch clamp electrophysiology has transformed research in the life sciences over the last few decades. Since their inception, automatic patch clamp platforms have evolved considerably, demonstrating the capability to address both voltage- and ligand-gated channels, and showing the potential to play a pivotal role in drug discovery and biomedical research. Unfortunately, the cell suspension assays to which early systems were limited cannot recreate biologically relevant cellular environments, or capture higher order aspects of synaptic physiology and network dynamics. In vivo patch clamp electrophysiology has the potential to yield more biologically complex information and be especially useful in reverse engineering the molecular and cellular mechanisms of single-cell and network neuronal computation, while capturing important aspects of human disease mechanisms and possible therapeutic strategies. Unfortunately, it is a difficult procedure with a steep learning curve, which has restricted dissemination of the technique. Luckily, in vivo patch clamp electrophysiology seems particularly amenable to robotic automation. In this review, we document the development of automated patch clamp technology, from early systems based on multi-well plates through to automated planar-array platforms, and modern robotic platforms capable of performing two-photon targeted whole-cell electrophysiological recordings in vivo.

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Haythornthwaite, Alison, Andrea Brueggemann, Cecilia Farre, Sonja Stoelzle, Claudia Haarmann, Michael George et Niels Fertig. « Automated patch clamp for hERG safety screening ». Journal of Pharmacological and Toxicological Methods 60, n^o 2 (septembre 2009) : 222. http://dx.doi.org/10.1016/j.vascn.2009.04.065.

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Jacobsen, Rasmus B., et Naja Møller M. Sørensen. « CFTR on an Automated Patch Clamp System ». Biophysical Journal 110, n^o 3 (février 2016) : 452a. http://dx.doi.org/10.1016/j.bpj.2015.11.2426.

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Holst, Gregory L., William Stoy, Bo Yang, Ilya Kolb, Suhasa B. Kodandaramaiah, Lu Li, Ulf Knoblich et al. « Autonomous patch-clamp robot for functional characterization of neurons in vivo : development and application to mouse visual cortex ». Journal of Neurophysiology 121, n^o 6 (1 juin 2019) : 2341–57. http://dx.doi.org/10.1152/jn.00738.2018.

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Patch clamping is the gold standard measurement technique for cell-type characterization in vivo, but it has low throughput, is difficult to scale, and requires highly skilled operation. We developed an autonomous robot that can acquire multiple consecutive patch-clamp recordings in vivo. In practice, 40 pipettes loaded into a carousel are sequentially filled and inserted into the brain, localized to a cell, used for patch clamping, and disposed. Automated visual stimulation and electrophysiology software enables functional cell-type classification of whole cell-patched cells, as we show for 37 cells in the anesthetized mouse in visual cortex (V1) layer 5. We achieved 9% yield, with 5.3 min per attempt over hundreds of trials. The highly variable and low-yield nature of in vivo patch-clamp recordings will benefit from such a standardized, automated, quantitative approach, allowing development of optimal algorithms and enabling scaling required for large-scale studies and integration with complementary techniques. NEW & NOTEWORTHY In vivo patch-clamp is the gold standard for intracellular recordings, but it is a very manual and highly skilled technique. The robot in this work demonstrates the most automated in vivo patch-clamp experiment to date, by enabling production of multiple, serial intracellular recordings without human intervention. The robot automates pipette filling, wire threading, pipette positioning, neuron hunting, break-in, delivering sensory stimulus, and recording quality control, enabling in vivo cell-type characterization.

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Bruhn, Brandon R., Haiyan Liu, Stefan Schuhladen, Alan J. Hunt, Aghapi Mordovanakis et Michael Mayer. « Dual-pore glass chips for cell-attached single-channel recordings ». Lab Chip 14, n^o 14 (2014) : 2410–17. http://dx.doi.org/10.1039/c4lc00370e.

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Wu (吴秋雨), Qiuyu, Ilya Kolb, Brendan M. Callahan, Zhaolun Su, William Stoy, Suhasa B. Kodandaramaiah, Rachael Neve et al. « Integration of autopatching with automated pipette and cell detection in vitro ». Journal of Neurophysiology 116, n^o 4 (1 octobre 2016) : 1564–78. http://dx.doi.org/10.1152/jn.00386.2016.

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Patch clamp is the main technique for measuring electrical properties of individual cells. Since its discovery in 1976 by Neher and Sakmann, patch clamp has been instrumental in broadening our understanding of the fundamental properties of ion channels and synapses in neurons. The conventional patch-clamp method requires manual, precise positioning of a glass micropipette against the cell membrane of a visually identified target neuron. Subsequently, a tight “gigaseal” connection between the pipette and the cell membrane is established, and suction is applied to establish the whole cell patch configuration to perform electrophysiological recordings. This procedure is repeated manually for each individual cell, making it labor intensive and time consuming. In this article we describe the development of a new automatic patch-clamp system for brain slices, which integrates all steps of the patch-clamp process: image acquisition through a microscope, computer vision-based identification of a patch pipette and fluorescently labeled neurons, micromanipulator control, and automated patching. We validated our system in brain slices from wild-type and transgenic mice expressing channelrhodopsin 2 under the Thy1 promoter (line 18) or injected with a herpes simplex virus-expressing archaerhodopsin, ArchT. Our computer vision-based algorithm makes the fluorescent cell detection and targeting user independent. Compared with manual patching, our system is superior in both success rate and average trial duration. It provides more reliable trial-to-trial control of the patching process and improves reproducibility of experiments.

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Harrison, Reid R., Ilya Kolb, Suhasa B. Kodandaramaiah, Alexander A. Chubykin, Aimei Yang, Mark F. Bear, Edward S. Boyden et Craig R. Forest. « Microchip amplifier for in vitro, in vivo, and automated whole cell patch-clamp recording ». Journal of Neurophysiology 113, n^o 4 (15 février 2015) : 1275–82. http://dx.doi.org/10.1152/jn.00629.2014.

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Patch clamping is a gold-standard electrophysiology technique that has the temporal resolution and signal-to-noise ratio capable of reporting single ion channel currents, as well as electrical activity of excitable single cells. Despite its usefulness and decades of development, the amplifiers required for patch clamping are expensive and bulky. This has limited the scalability and throughput of patch clamping for single-ion channel and single-cell analyses. In this work, we have developed a custom patch-clamp amplifier microchip that can be fabricated using standard commercial silicon processes capable of performing both voltage- and current-clamp measurements. A key innovation is the use of nonlinear feedback elements in the voltage-clamp amplifier circuit to convert measured currents into logarithmically encoded voltages, thereby eliminating the need for large high-valued resistors, a factor that has limited previous attempts at integration. Benchtop characterization of the chip shows low levels of current noise [1.1 pA root mean square (rms) over 5 kHz] during voltage-clamp measurements and low levels of voltage noise (8.2 μV rms over 10 kHz) during current-clamp measurements. We demonstrate the ability of the chip to perform both current- and voltage-clamp measurement in vitro in HEK293FT cells and cultured neurons. We also demonstrate its ability to perform in vivo recordings as part of a robotic patch-clamping system. The performance of the patch-clamp amplifier microchip compares favorably with much larger commercial instrumentation, enabling benchtop commoditization, miniaturization, and scalable patch-clamp instrumentation.

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P.G. Korsgaard, Mads. « Voltage- and Current Clamp on Induced Pluripotent Cardiomyocytes with Automated Patch Clamp ». Biophysical Journal 112, n^o 3 (février 2017) : 414a. http://dx.doi.org/10.1016/j.bpj.2016.11.2218.

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Brüggemann, Andrea, Claudia Haarmann, Markus Rapedius, Tom Goetze, Ilka Rinke, Michael George et Niels Fertig. « Characterization of iPS Derived Cardiomyocytes in Voltage Clamp and Current Clamp by Automated Patch Clamp ». Biophysical Journal 112, n^o 3 (février 2017) : 236a. http://dx.doi.org/10.1016/j.bpj.2016.11.1290.

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Haythornthwaite, Alison, Sonja Stoelzle, Alexander Hasler, Andrea Kiss, Johannes Mosbacher, Michael George, Andrea Brüggemann et Niels Fertig. « Characterizing Human Ion Channels in Induced Pluripotent Stem Cell–Derived Neurons ». Journal of Biomolecular Screening 17, n^o 9 (24 août 2012) : 1264–72. http://dx.doi.org/10.1177/1087057112457821.

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Neurons derived from human-induced pluripotent stem cells were characterized using manual and automated patch-clamp recordings. These cells expressed voltage-gated Na+ (Nav), Ca2+ (Cav), and K+ (Kv) channels as expected from excitable cells. The Nav current was TTX sensitive, IC50 = 12 ± 6 nM ( n = 5). About 50% of the Cav current was blocked by 10 µM of the L-type channel blocker nifedipine. Two populations of the Kv channel were present in different proportions: an inactivating (A-type) and a noninactivating type. The A-type current was sensitive to 4-AP and TEA (IC50 = 163 ± 93 µM; n = 3). Application of γ-aminobutyric acid (GABA) activated a current sensitive to the GABAA receptor antagonist bicuculline, IC50 = 632 ± 149 nM ( n = 5). In both devices, comparable action potentials were generated in the current clamp. With unbiased, automated patch clamp, about 40% of the cells expressed Nav currents, whereas visual guidance in manual patch clamp provided almost a 100% success rate of patching “excitable cells.” These results show high potential for pluripotent stem cell–derived neurons as a useful model for drug discovery, in combination with automated patch-clamp recordings for high-throughput and high-quality drug assessments at human neuronal ion channels in their correct cellular background.

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Golubovic, Andjelko, Martina Knirsch, Uwe Czubayko, Alexander Hummer, Albrecht Lepple-Wienhues et Dirck Lassen. « The Chiptip : A Novel Tool for Automated Patch Clamp ». Combinatorial Chemistry & ; High Throughput Screening 12, n^o 1 (1 janvier 2009) : 73–77. http://dx.doi.org/10.2174/138620709787047984.

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Humphries, Edward, John Ridley, Robert Kirby et Marc Rogers. « Predicting proarrhythmic risk exclusively using automated patch clamp data ». Journal of Pharmacological and Toxicological Methods 105 (septembre 2020) : 106728. http://dx.doi.org/10.1016/j.vascn.2020.106728.

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Haythornthwaite, Alison, Andrea Brueggemann, Cecilia Farre, Sonja Stoelzle, Claudia Haarmann, Michael George et Niels Fertig. « Automated patch clamp for temperature controlled hERG compound screening ». Journal of Pharmacological and Toxicological Methods 58, n^o 2 (septembre 2008) : 155–56. http://dx.doi.org/10.1016/j.vascn.2008.05.043.

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Brüggemann, Andrea, Sonja Stoelzle, Michael George, Jan C Behrends et Niels Fertig. « Microchip Technology for Automated and Parallel Patch-Clamp Recording ». Small 2, n^o 7 (juillet 2006) : 840–46. http://dx.doi.org/10.1002/smll.200600083.

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Farre, Cecilia, Michael George, Andrea Brüggemann et Niels Fertig. « Ion channel screening – automated patch clamp on the rise ». Drug Discovery Today : Technologies 5, n^o 1 (mars 2008) : e23-e28. http://dx.doi.org/10.1016/j.ddtec.2008.12.004.

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Barthmes, Maria, Mac Donald F. Jose, Jan Peter Birkner, Andrea Brüggemann, Christian Wahl-Schott et Armağan Koçer. « Studying mechanosensitive ion channels with an automated patch clamp ». European Biophysics Journal 43, n^o 2-3 (14 février 2014) : 97–104. http://dx.doi.org/10.1007/s00249-014-0944-2.

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Hasan, Md Mahadhi, Lotten Ragnarsson, Fernanda C. Cardoso et Richard J. Lewis. « Transfection methods for high-throughput cellular assays of voltage-gated calcium and sodium channels involved in pain ». PLOS ONE 16, n^o 3 (5 mars 2021) : e0243645. http://dx.doi.org/10.1371/journal.pone.0243645.

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Chemical transfection is broadly used to transiently transfect mammalian cells, although often associated with cellular stress and membrane instability, which imposes challenges for most cellular assays, including high-throughput (HT) assays. In the current study, we compared the effectiveness of calcium phosphate, FuGENE and Lipofectamine 3000 to transiently express two key voltage-gated ion channels critical in pain pathways, CaV2.2 and NaV1.7. The expression and function of these channels were validated using two HT platforms, the Fluorescence Imaging Plate Reader FLIPRTetra and the automated patch clamp QPatch 16X. We found that all transfection methods tested demonstrated similar effectiveness when applied to FLIPRTetra assays. Lipofectamine 3000-mediated transfection produced the largest peak currents for automated patch clamp QPatch assays. However, the FuGENE-mediated transfection was the most effective for QPatch assays as indicated by the superior number of cells displaying GΩ seal formation in whole-cell patch clamp configuration, medium to large peak currents, and higher rates of accomplished assays for both CaV2.2 and NaV1.7 channels. Our findings can facilitate the development of HT automated patch clamp assays for the discovery and characterization of novel analgesics and modulators of pain pathways, as well as assisting studies examining the pharmacology of mutated channels.

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Lei, Chon Lok, Alan Fabbri, Dominic G. Whittaker, Michael Clerx, Monique J. Windley, Adam P. Hill, Gary R. Mirams et Teun P. de Boer. « A nonlinear and time-dependent leak current in the presence of calcium fluoride patch-clamp seal enhancer ». Wellcome Open Research 5 (2 novembre 2021) : 152. http://dx.doi.org/10.12688/wellcomeopenres.15968.2.

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Automated patch-clamp platforms are widely used and vital tools in both academia and industry to enable high-throughput studies such as drug screening. A leak current to ground occurs whenever the seal between a pipette and cell (or internal solution and cell in high-throughput machines) is not perfectly insulated from the bath (extracellular) solution. Over 1 GΩ seal resistance between pipette and bath solutions is commonly used as a quality standard for manual patch work. With automated platforms it can be difficult to obtain such a high seal resistance between the intra- and extra-cellular solutions. One suggested method to alleviate this problem is using an F− containing internal solution together with a Ca2+ containing external solution — so that a CaF2 crystal forms when the two solutions meet which ‘plugs the holes’ to enhance the seal resistance. However, we observed an unexpected nonlinear-in-voltage and time-dependent current using these solutions on an automated patch-clamp platform. We performed manual patch-clamp experiments with the automated patch-clamp solutions, but no biological cell, and observed the same nonlinear time-dependent leak current. The current could be completely removed by washing out F− ions to leave a conventional leak current that was linear and not time-dependent. We therefore conclude fluoride ions interacting with the CaF2 crystal are the origin of the nonlinear time-dependent leak current. The consequences of such a nonlinear and time-dependent leak current polluting measurements should be considered carefully if it cannot be isolated and subtracted.

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Lei, Chon Lok, Alan Fabbri, Dominic G. Whittaker, Michael Clerx, Monique J. Windley, Adam P. Hill, Gary R. Mirams et Teun P. de Boer. « A nonlinear and time-dependent leak current in the presence of calcium fluoride patch-clamp seal enhancer ». Wellcome Open Research 5 (1 juillet 2020) : 152. http://dx.doi.org/10.12688/wellcomeopenres.15968.1.

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Automated patch-clamp platforms are widely used and vital tools in both academia and industry to enable high-throughput studies such as drug screening. A leak current to ground occurs whenever the seal between a pipette and cell (or internal solution and cell in high-throughput machines) is not perfectly insulated from the bath (extracellular) solution. Over 1 GΩ seal resistance between pipette and bath solutions is commonly used as a quality standard for manual patch work. With automated platforms it can be difficult to obtain such a high seal resistance between the intra- and extra-cellular solutions. One suggested method to alleviate this problem is using an F− containing internal solution together with a Ca2+ containing external solution — so that a CaF2 crystal forms when the two solutions meet which ‘plugs the holes’ to enhance the seal resistance. However, we observed an unexpected nonlinear-in-voltage and time-dependent current using these solutions on an automated patch-clamp platform. We performed manual patch-clamp experiments with the automated patch-clamp solutions, but no biological cell, and observed the same nonlinear time-dependent leak current. The current could be completely removed by washing out F− ions to leave a conventional leak current that was linear and not time-dependent. We therefore conclude fluoride ions interacting with the CaF2 crystal are the origin of the nonlinear time-dependent leak current. The consequences of such a nonlinear and time-dependent leak current polluting measurements should be considered carefully if it cannot be isolated and subtracted.

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Obergrussberger, A., A. Bru ggemann, T. A. Goetze, M. Rapedius, C. Haarmann, I. Rinke, N. Becker et al. « Automated Patch Clamp Meets High-Throughput Screening : 384 Cells Recorded in Parallel on a Planar Patch Clamp Module ». Journal of Laboratory Automation 21, n^o 6 (23 décembre 2015) : 779–93. http://dx.doi.org/10.1177/2211068215623209.

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Desai, Niraj S., Jennifer J. Siegel, William Taylor, Raymond A. Chitwood et Daniel Johnston. « MATLAB-based automated patch-clamp system for awake behaving mice ». Journal of Neurophysiology 114, n^o 2 (août 2015) : 1331–45. http://dx.doi.org/10.1152/jn.00025.2015.

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Automation has been an important part of biomedical research for decades, and the use of automated and robotic systems is now standard for such tasks as DNA sequencing, microfluidics, and high-throughput screening. Recently, Kodandaramaiah and colleagues ( Nat Methods 9: 585–587, 2012) demonstrated, using anesthetized animals, the feasibility of automating blind patch-clamp recordings in vivo. Blind patch is a good target for automation because it is a complex yet highly stereotyped process that revolves around analysis of a single signal (electrode impedance) and movement along a single axis. Here, we introduce an automated system for blind patch-clamp recordings from awake, head-fixed mice running on a wheel. In its design, we were guided by 3 requirements: easy-to-use and easy-to-modify software; seamless integration of behavioral equipment; and efficient use of time. The resulting system employs equipment that is standard for patch recording rigs, moderately priced, or simple to make. It is written entirely in MATLAB, a programming environment that has an enormous user base in the neuroscience community and many available resources for analysis and instrument control. Using this system, we obtained 19 whole cell patch recordings from neurons in the prefrontal cortex of awake mice, aged 8–9 wk. Successful recordings had series resistances that averaged 52 ± 4 MΩ and required 5.7 ± 0.6 attempts to obtain. These numbers are comparable with those of experienced electrophysiologists working manually, and this system, written in a simple and familiar language, will be useful to many cellular electrophysiologists who wish to study awake behaving mice.

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Stoelzle-Feix, Sonja, Andras Horvath, Nadine Becker, Alan Fabbri, Christian Grad, Michael George, Teun P. de Boer et Niels Fertig. « Automated patch clamp system introducing simulated Ik1 into human iPSC-cardiomycoytes using dynamic clamp ». Journal of Pharmacological and Toxicological Methods 105 (septembre 2020) : 106744. http://dx.doi.org/10.1016/j.vascn.2020.106744.

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Stoelzle, Sonja, Alison Haythornthwaite, Claudia Haarmann, Cecilia Farre, Rodolfo Haedo, Andrea Brüggemann, Michael George et Niels Fertig. « Automated Patch Clamp with Current Clamp : Action Potential Recordings from Stem Cell Derived Cardiomyocytes ». Biophysical Journal 100, n^o 3 (février 2011) : 196a. http://dx.doi.org/10.1016/j.bpj.2010.12.1287.

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Stoelzle, Sonja, Alison Haythornthwaite, Ralf Kettenhofen, Eugen Kolossov, Heribert Bohlen, Michael George, Andrea Brüggemann et Niels Fertig. « Automated Patch Clamp on mESC-Derived Cardiomyocytes for Cardiotoxicity Prediction ». Journal of Biomolecular Screening 16, n^o 8 (20 juillet 2011) : 910–16. http://dx.doi.org/10.1177/1087057111413924.

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Cardiovascular side effects are critical in drug development and have frequently led to late-stage project terminations or even drug withdrawal from the market. Physiologically relevant and predictive assays for cardiotoxicity are hence strongly demanded by the pharmaceutical industry. To identify a potential impact of test compounds on ventricular repolarization, typically a variety of ion channels in diverse heterologously expressing cells have to be investigated. Similar to primary cells, in vitro–generated stem cell–derived cardiomyocytes simultaneously express cardiac ion channels. Thus, they more accurately represent the native situation compared with cell lines overexpressing only a single type of ion channel. The aim of this study was to determine if stem cell–derived cardiomyocytes are suited for use in an automated patch clamp system. The authors show recordings of cardiac ion currents as well as action potential recordings in readily available stem cell–derived cardiomyocytes. Besides monitoring inhibitory effects of reference compounds on typical cardiac ion currents, the authors revealed for the first time drug-induced modulation of cardiac action potentials in an automated patch clamp system. The combination of an in vitro cardiac cell model with higher throughput patch clamp screening technology allows for a cost-effective cardiotoxicity prediction in a physiologically relevant cell system.

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Kodandaramaiah, Suhasa B., Giovanni Talei Franzesi, Brian Y. Chow, Edward S. Boyden et Craig R. Forest. « Automated whole-cell patch-clamp electrophysiology of neurons in vivo ». Nature Methods 9, n^o 6 (6 mai 2012) : 585–87. http://dx.doi.org/10.1038/nmeth.1993.

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Danker, Timm, Manfred Frey et Elke Guenther. « Inducible CaV1.2 cell line optimized for automated patch clamp assays ». Journal of Pharmacological and Toxicological Methods 81 (septembre 2016) : 341. http://dx.doi.org/10.1016/j.vascn.2016.02.023.

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Stoelzle, Sonja, Andrea Bruggemann, David Guinot, Alison Haythornthwaite, Michael George, Cecilia Farre, Claudia Haarmann, Ralf Kettenhofen et Niels Fertig. « Excitement Over Automated Patch Clamp : Action Potentials from Cardiac Myocytes ». Biophysical Journal 98, n^o 3 (janvier 2010) : 514a. http://dx.doi.org/10.1016/j.bpj.2009.12.2798.

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Schrøder, Rikke L., Mette Christensen, Blake Anson et Morten Sunesen. « Exploring Stem Cell-Derived Cardiomyocytes with Automated Patch Clamp Techniques ». Biophysical Journal 102, n^o 3 (janvier 2012) : 544a. http://dx.doi.org/10.1016/j.bpj.2011.11.2968.

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Mathes, Chris. « QPatch : the past, present and future of automated patch clamp ». Expert Opinion on Therapeutic Targets 10, n^o 2 (21 mars 2006) : 319–27. http://dx.doi.org/10.1517/14728222.10.2.319.

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Mike, Arpad, Krisztina Pesti, Mátyás Csaba Földi, Zsolt Bagoly, Gábor Papp et Péter Lukács. « Optimizing for Information Content on Ionflux Mercury Automated Patch Clamp ». Biophysical Journal 118, n^o 3 (février 2020) : 590a—591a. http://dx.doi.org/10.1016/j.bpj.2019.11.3199.

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Okeyo, George O., András Horváth, Nadine Becker, Alan Fabbri, Christian Grad, Michael George, Teun P. de Boer et Niels Fertig. « Automated Patch Clamp System Introducing Simulated IK1 into Stem Cell-Derived Cardiomyocytes using Dynamic Clamp ». Biophysical Journal 118, n^o 3 (février 2020) : 571a. http://dx.doi.org/10.1016/j.bpj.2019.11.3105.

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Vanoye, Carlos G., Reshma R. Desai, Nirvani Jairam, Nora Ghabra, Jens Meiler, Charles R. Sanders et Alfred L. George. « Comprehensive functional evaluation of KCNE1 variants using automated patch clamp recording ». Biophysical Journal 121, n^o 3 (février 2022) : 241a. http://dx.doi.org/10.1016/j.bpj.2021.11.1547.

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Brinkwirth, Nina, Nadine Becker, Alison Obergrussberger, Rodolfo Haedo, Claudia Haarmann, Niels Fertig et Sonja Stoelzle-Feix. « Unattended screening workflow in a 384-well automated patch clamp system ». Journal of Pharmacological and Toxicological Methods 111 (septembre 2021) : 106970. http://dx.doi.org/10.1016/j.vascn.2021.106970.

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Balasubramanian, Bharathi, John P. Imredy, David Kim, Jacob Penniman, Armando Lagrutta et Joseph J. Salata. « Optimization of Cav1.2 screening with an automated planar patch clamp platform ». Journal of Pharmacological and Toxicological Methods 59, n^o 2 (mars 2009) : 62–72. http://dx.doi.org/10.1016/j.vascn.2009.02.002.

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Zhao, Yong, Beth Hooten et Lori Jin. « Screening of hERG blockers by automated patch clamp system : QPatch 16 ». Journal of Pharmacological and Toxicological Methods 60, n^o 2 (septembre 2009) : 223. http://dx.doi.org/10.1016/j.vascn.2009.04.066.

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Haedo, Rodolfo J. « Safety pharmacology measurements within the CiPA initiative using automated patch clamp ». Journal of Pharmacological and Toxicological Methods 99 (septembre 2019) : 106595. http://dx.doi.org/10.1016/j.vascn.2019.05.024.

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Johnson, Juliette. « Nicotinic Receptors Investigated Using a Next Generation Automated Patch Clamp System ». Biophysical Journal 100, n^o 3 (février 2011) : 274a. http://dx.doi.org/10.1016/j.bpj.2010.12.1703.

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Ionescu-Zanetti, Cristian, et Qin Chen. « Temperature Dependence of Herg Blocker Pharmacology - an Automated Patch Clamp Study ». Biophysical Journal 100, n^o 3 (février 2011) : 573a. http://dx.doi.org/10.1016/j.bpj.2010.12.3319.

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Chevalier, Morgan, Bogdan Amuzescu, Vaibhavkumar Gawali, Hannes Todt, Thomas Knott, Olaf Scheel et Hugues Abriel. « Late cardiac sodium current can be assessed using automated patch-clamp ». F1000Research 3 (16 octobre 2014) : 245. http://dx.doi.org/10.12688/f1000research.5544.1.

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The cardiac late Na+ current is generated by a small fraction of voltage-dependent Na+ channels that undergo a conformational change to a burst-gating mode, with repeated openings and closures during the action potential (AP) plateau. Its magnitude can be augmented by inactivation-defective mutations, myocardial ischemia, or prolonged exposure to chemical compounds leading to drug-induced (di)-long QT syndrome, and results in an increased susceptibility to cardiac arrhythmias. Using CytoPatch™ 2 automated patch-clamp equipment, we performed whole-cell recordings in HEK293 cells stably expressing human Nav1.5, and measured the late Na+ component as average current over the last 100 ms of 300 ms depolarizing pulses to -10 mV from a holding potential of -100 mV, with a repetition frequency of 0.33 Hz. Averaged values in different steady-state experimental conditions were further corrected by the subtraction of current average during the application of tetrodotoxin (TTX) 30 μM. We show that ranolazine at 10 and 30 μM in 3 min applications reduced the late Na+ current to 75.0 ± 2.7% (mean ± SEM, n = 17) and 58.4 ± 3.5% (n = 18) of initial levels, respectively, while a 5 min application of veratridine 1 μM resulted in a reversible current increase to 269.1 ± 16.1% (n = 28) of initial values. Using fluctuation analysis, we observed that ranolazine 30 μM decreased mean open probability p from 0.6 to 0.38 without modifying the number of active channels n, while veratridine 1 μM increased n 2.5-fold without changing p. In human iPSC-derived cardiomyocytes, veratridine 1 μM reversibly increased APD90 2.12 ± 0.41-fold (mean ± SEM, n = 6). This effect is attributable to inactivation removal in Nav1.5 channels, since significant inhibitory effects on hERG current were detected at higher concentrations in hERG-expressing HEK293 cells, with a 28.9 ± 6.0% inhibition (mean ± SD, n = 10) with 50 μM veratridine.

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Rapedius, Markus, Andrea Bruggemann, Tom Goetze, Claudia Haarmann, Ilka Rinke, Atsushi Ohtsuki, Takayuki Oka et al. « Characterization of NaV1.8 on a Highly Parallel Automated Patch Clamp System ». Biophysical Journal 110, n^o 3 (février 2016) : 113a. http://dx.doi.org/10.1016/j.bpj.2015.11.662.

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Schrøder, Rikke L., Søren Friis, Morten Sunesen, Chris Mathes et Niels J. Willumsen. « Automated Patch-Clamp Technique : Increased Throughput in Functional Characterization and in Pharmacological Screening of Small-Conductance Ca2+ Release-Activated Ca2+ Channels ». Journal of Biomolecular Screening 13, n^o 7 (1 juillet 2008) : 638–47. http://dx.doi.org/10.1177/1087057108320274.

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The suitability of an automated patch clamp for the characterization and pharmacological screening of calcium release—activated calcium (CRAC) channels endogenously expressed in RBL-2H3 cells was explored with the QPatch system. CRAC currents (I CRAC) are small, and thus precise recordings require high signal-to-noise ratios obtained by high seal resistances. Automated whole-cell establishment resulted in membrane resistances of 1728 ± 226 MΩ ( n = 44). CRAC channels were activated by a number of methods that raise intracellular calcium concentration, including EGTA, ionomycin, Ins(1,4,5)P3, and thapsigargin. ICRAC whole-cell currents ranged from 30 to 120 pA with rise times of 40 to 150 s. An initial delay in current activation was observed in particular when ICRAC was activated by passive store depletion using EGTA. Apparent rundown of ICRAC was commonly observed, and the current could be reactivated by subsequent addition of thapsigargin. ICRAC was blocked by SKF-96365 and 2-APB with IC50 values of 4.7 ± 1.1 µM ( n = 9) and 7.5 ± 0.7 ( n = 9) µM, respectively. The potencies of these blockers were similar to values reported for ICRAC in similar conventional patch-clamp experiments. The study demonstrates that CRAC channels can be rapidly and efficiently targeted with automated patch-clamp techniques for characterization of physiological and pharmacological properties. ( Journal of Biomolecular Screening 2008:638-647)

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Bettini, Ezio, Stephen M. Stahl, Sara De Martin, Andrea Mattarei, Jacopo Sgrignani, Corrado Carignani, Selena Nola et al. « Pharmacological Comparative Characterization of REL-1017 (Esmethadone-HCl) and Other NMDAR Channel Blockers in Human Heterodimeric N-Methyl-D-Aspartate Receptors ». Pharmaceuticals 15, n^o 8 (13 août 2022) : 997. http://dx.doi.org/10.3390/ph15080997.

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Excessive Ca2+ currents via N-methyl-D-aspartate receptors (NMDARs) have been implicated in many disorders. Uncompetitive NMDAR channel blockers are an emerging class of drugs in clinical use for major depressive disorder (MDD) and other neuropsychiatric diseases. The pharmacological characterization of uncompetitive NMDAR blockers in clinical use may improve our understanding of NMDAR function in physiology and pathology. REL-1017 (esmethadone-HCl), a novel uncompetitive NMDAR channel blocker in Phase 3 trials for the treatment of MDD, was characterized together with dextromethorphan, memantine, (±)-ketamine, and MK-801 in cell lines over-expressing NMDAR subtypes using fluorometric imaging plate reader (FLIPR), automated patch-clamp, and manual patch-clamp electrophysiology. In the absence of Mg2+, NMDAR subtypes NR1-2D were most sensitive to low, sub-μM glutamate concentrations in FLIPR experiments. FLIPR Ca2+ determination demonstrated low μM affinity of REL-1017 at NMDARs with minimal subtype preference. In automated and manual patch-clamp electrophysiological experiments, REL-1017 exhibited preference for the NR1-2D NMDAR subtype in the presence of 1 mM Mg2+ and 1 μM L-glutamate. Tau off and trapping characteristics were similar for (±)-ketamine and REL-1017. Results of radioligand binding assays in rat cortical neurons correlated with the estimated affinities obtained in FLIPR assays and in automated and manual patch-clamp assays. In silico studies of NMDARs in closed and open conformation indicate that REL-1017 has a higher preference for docking and undocking the open-channel conformation compared to ketamine. In conclusion, the pharmacological characteristics of REL-1017 at NMDARs, including relatively low affinity at the NMDAR, NR1-2D subtype preference in the presence of 1 mM Mg2+, tau off and degree of trapping similar to (±)-ketamine, and preferential docking and undocking of the open NMDAR, could all be important variables for understanding the rapid-onset antidepressant effects of REL-1017 without psychotomimetic side effects.

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Stoelzle-Feix, Sonja, Nadine Becker, Alison Obergrussberger, Teun de Boer, Birgit Goversen, Toon A. van Veen et Niels Fertig. « Introducing simulated IK1 into human iPSC-cardiomyocytes using dynamic clamp on an automated patch clamp system ». Journal of Pharmacological and Toxicological Methods 99 (septembre 2019) : 106595. http://dx.doi.org/10.1016/j.vascn.2019.05.040.

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Bot, Corina, Nadine Becker, Birgit Goversen, Sonja Stoelzle-Feix, Alison Obergrussberger, Toon A. B. van Veen, Niels Fertig et Teun P. de Boer. « Introducing Simulated IK1 into Human iPSC-Cardiomyocytes using Dynamic Clamp on an Automated Patch Clamp Platform ». Biophysical Journal 114, n^o 3 (février 2018) : 308a. http://dx.doi.org/10.1016/j.bpj.2017.11.1747.

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Lu, Gang, András Horváth, Nadine Becker, Alan Fabbr, Christian Grad, Michael George, Niels Fertig et Teun P. de Boer. « Introducing Simulated IK1 into Human iPSC-Cardiomyocytes using Dynamic Clamp on an Automated Patch Clamp System ». Biophysical Journal 116, n^o 3 (février 2019) : 99a. http://dx.doi.org/10.1016/j.bpj.2018.11.572.

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Boussaoud, Adrien, et Nathalie Picollet-D'Hahan. « Toxint’patch : a briefcase-sized system for toxin detection using planar patch-clamp. Validation of an integrated and automated patch clamp device ». Instrumentation Mesure Métrologie 11, n^o 1-2 (30 juin 2011) : 9–29. http://dx.doi.org/10.3166/i2m.11.1-2.9-29.

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Li, Chenhong. « A reliable whole cell clamp technique ». Advances in Physiology Education 32, n^o 3 (septembre 2008) : 209–11. http://dx.doi.org/10.1152/advan.90138.2008.

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This article describes a simple whole cell formation technique that the author invented in teaching and experiments. The implementation of the invented technique is a syringe with a hole and slot. With the newly invented technique, novices will shorten their learning curve and veterans will increase their success rate. The invented technique lightens the labor of the experimenter and improves the success rate and quality of whole cell preparations. The article also provides an idea to design an automated whole cell formation implementation. The tools developed in this technique make the patch-clamp experiment easy to teach and learn.

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Rubaiy, Hussein Nori. « A Short Guide to Electrophysiology and Ion Channels ». Journal of Pharmacy & ; Pharmaceutical Sciences 20 (15 mars 2017) : 48. http://dx.doi.org/10.18433/j32p6r.

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The birth and discovery of electrophysiological science took place in the 18th century laying the path for our understanding of nerve membrane ionic currents. The pore-forming proteins, ion channels, are involved and play critical roles in very important physiological and pathological processes, such as neuronal signaling and cardiac excitability, therefore, they serve as therapeutic drug targets. The study of physiological, pharmacological and biophysical properties of ion channels can be done by patch clamp, a gold standard and powerful electrophysiological technique. The current review, in addition to highlight and cover the history of electrophysiology, patch clamp (conventional and automated) technique, and different types of ion channels, will also discuss the importance of ion channels in different neurological diseases and disorders. As the field of neuroscience is growing, this manuscript is intended as a guide to help in understanding the importance of ion channels, particularly in neuroscience, and also in using the patch clamp technique for the study of molecular physiology, pathophysiology, and pharmacology of neuronal ion channels. Importantly, this review will spotlight on the therapeutic aspect of neuronal ion channels. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.

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Lepple-Wienhues, Albrecht, Klaus Ferlinz, Achim Seeger et Arvid Schäfer. « Flip the Tip : An Automated, High Quality, Cost-Effective Patch Clamp Screen ». Receptors and Channels 9, n^o 1 (janvier 2003) : 13–17. http://dx.doi.org/10.3109/10606820308257.

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