Pclamp 10

Manufactured by Molecular Devices

Sourced in United States, Germany, Canada, United Kingdom, China, Australia

PClamp 10 software is a data acquisition and analysis platform for electrophysiology research. It provides tools for recording, analyzing, and visualizing electrical signals from cells and tissues.

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Lab products found in correlation

1 101 protocols using pclamp 10

Electrophysiological Recordings and Analysis

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Data were transferred to a computer hard disk after digitization with an A/D converter (Digidata 1550, Molecular Devices). Data acquisition (digitized at 10 kHz and filtered at 3 kHz) was performed with pClamp 10.4 software (Molecular Devices, Sunnyvale, CA, USA). Input resistance and cells capacitance were measured online with the membrane test feature of the pClamp software. Spontaneous EPSCs and IPSCs were analyzed with pClamp 10.4 (Molecular Devices, Sunnyvale, CA, USA). This program uses a detection algorithm based on a sliding template. The template did not induce any bias in the sampling of events because it was moved along the data trace by one point at a time and was optimally scaled to fit the data at each position.
The rise time of the evoked IPSC was estimated as the time needed for 20–80% increase of the peak current response. The decay time was fitted with first order exponential function in the following form:
where τ and A are the time constants and relative fractions of the respective components.

Modi B., Pimpinella D., Pazienti A., Zacchi P., Cherubini E, & Griguoli M. (2019). Possible Implication of the CA2 Hippocampal Circuit in Social Cognition Deficits Observed in the Neuroligin 3 Knock-Out Mouse, a Non-Syndromic Animal Model of Autism. Frontiers in Psychiatry, 10, 513.

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Electrophysiological Data Acquisition and Analysis

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Data were transferred to a computer hard disk after digitization with an A/D converter (Digidata 1550, Molecular Devices, Sunnyvale, CA, USA). Data acquisition (digitized at 10 kHz and filtered at 3 kHz) was performed with pClamp 10.4 software (Molecular Devices, Sunnyvale, CA, USA). Input resistance and cells capacitance were measured online with the membrane test feature of the pClamp software. Spontaneous and miniature EPSCs and IPSCs were analyzed with pClamp 10.4 (Molecular Devices, Sunnyvale, CA, USA). This program uses a detection algorithm based on a sliding template. The template did not induce any bias in the sampling of events because it was moved along the data trace by one point at a time and was optimally scaled to fit the data at each position.

Pimpinella D., Mastrorilli V., Giorgi C., Coemans S., Lecca S., Lalive A.L., Ostermann H., Fuchs E.C., Monyer H., Mele A., Cherubini E, & Griguoli M. (2021). Septal cholinergic input to CA2 hippocampal region controls social novelty discrimination via nicotinic receptor-mediated disinhibition. eLife, 10, e65580.

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Measuring RyR1 channel modulation by HIV drugs

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For this, phosphatidylethanolamine, phosphatidylserine and phosphatidylcholine in a ratio of 5:3:2 (35 mg/mL of lipid) in n-decane were painted across a 250 μm diameter hole of the bilayer cup as described earlier [31 (link),36 (link),37 (link)]. Proteoliposomes containing purified RyR1 was then added to the cis side (equivalent to the cytoplasm) to allow a single RyR1 to fuse into the bilayer. The other side of the bilayer cup was designated the trans side (equivalent to the lumen of the SR). All recordings were performed in symmetric KCl buffer solutions (0.25 mM KCl, 20 mM K/HEPES, pH 7.4) with the cis chamber containing 3.0 – 3.3 μM Ca²⁺. EFV (0 – 31.2 μM), ATV (0 – 30 μM) and RTV (0 – 30 μM) were added sequentially as boluses to the cis chamber at 100 times higher than the final concentrations and the chamber were vigorously stirred for 15 s prior to obtaining a 2 min recording at ± 30 mV. All experiments were conducted at room temperature (23–25°C) and electrical signals were filtered at 2 kHz and digitized at 10 kHz. Data were acquired using commercially available equipment and software (Axopatch 1D, Digidata 1322A and pClamp 10.2, Axon Instruments, Burlingame, CA, USA). Data were analyzed using pClamp 10.2 (Axon Instruments, Burlingame, CA, USA), Sigma Plot 10.0 (Stystat Software Inc, Chicago, IL, USA) and GraphPad Prism (Ver 7).

Alomar F.A., Tian C., Dash P.K., McMillan J.M., Gendelman H.E., Gorantla S, & Bidasee K.R. (2021). EFAVIRENZ, ATAZANAVIR, AND RITONAVIR DISRUPT SARCOPLASMIC RETICULUM Ca2+ HOMEOSTASIS IN SKELETAL MUSCLES. Antiviral research, 187, 104975.

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Analyzing Neuromuscular Function in Drosophila Larvae

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The third instar larvae were dissected in 0 mM Ca²⁺ HL-3 at room temperature and then bathed in 1 mM Ca²⁺ HL-3 solution for 5–10 min before the recording. The mean value of the resistance of the recording electrode was ~40 MΩ when the electrode was filled with a 3M KCl solution. All recordings were obtained from muscle 6 of abdominal segment 3. Each larva was only used for one recording. Recordings from the muscles that hold resting membrane potentials at less than −60 mV were used for further data quantifications. EJPs were evoked by stimulating the axonal bundle via a glass capillary electrode with an internal diameter of ~10–15 μm (Harvard Apparatus Glass Capillaries GC120F-15) at 0.2 Hz. Stimulus pulses were fixed at 0.5 ms duration (pClamp 10.6 software, Axon Instruments). To obtain maximal EJP amplitude, 3–5 mV electric stimuli were applied. EJPs were amplified with an Axoclamp 900A amplifier (Axon Instruments, Foster City, California) under bridge mode and filtered at 10 kHz. EJPs were analyzed by pClamp 10.6 software (Axon Instruments). For the EJP amplitude at 0.2 Hz, the mean of the EJP amplitude was averaged from the amplitudes of 80 EJPs in one consecutive recording.

Yao C.K., Liu Y.T., Lee I.C., Wang Y.T, & Wu P.Y. (2017). A Ca2+ channel differentially regulates Clathrin-mediated and activity-dependent bulk endocytosis. PLoS Biology, 15(4), e2000931.

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Patch-Clamp Recording of hERG Currents

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For electrophysiological recording, the cover glasses containing adherent hERG-HEK293 recombinant cells were transferred to the recording chamber (RC-13, Warner Instruments) mounted on the stage of an inverted microscope (IX70, Olympus, Tokyo, Japan). Cells were continuously perfused with an external bath solution. hERG currents were recorded using a Multiclamp 700 B microelectrode amplifier and pClamp 10.1 software (Molecular Devices, Sunnyvale, CA, USA) in a whole-cell configuration of the patch-clamp technique at room temperature (22~24℃). Glass micropipettes were pulled from glass capillaries (PG10165-4, World Precision Instruments, Sarasota, FL, USA) using a programmable horizontal microelectrode puller (P-97, Sutter Instrument, Novato, CA, USA). The tip resistances of the patch pipettes were 2~4 MΩ when filled with the internal solution. The liquid junction potentials between pipette and bath solutions were in the range 3~5 mV, and zeroed before the gigaohm seal was formed. The current signals were filtered at 2 kHz, digitized at 10 kHz, and saved on a PC using DigiData 1322 and pClamp 10.1 software (Molecular Devices).

Joo Y.S., Lee H.J., Choi J.S, & Sung K.W. (2016). Acepromazine inhibits hERG potassium ion channels expressed in human embryonic kidney 293 cells. The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology, 21(1), 75-82.

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Whole-cell Patch Clamp Recordings in Acute Brain Slices

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Slices were transferred to a recording chamber and were continuously perfused with oxygenated Tyrode’s solution at 32°C - 34°C which included synaptic blockers SR95531 (gabazine, 10μM), 6,7-dinitroquinoxaline-2,3-dione (DNQX, 10μM), and (R)-CPP (10μM). All drugs were purchased from Tocris (Bristol, United Kingdom). Glass patch pipettes (Sutter Instruments BF150–86-7.5) were pulled using a Sutter P97 (Sutter Instrument, Novato, CA, USA) to produce tip resistances of 2 – 4 MΩ and were filled with a K-gluconate intracellular solution (290 mOsm, pH 7.37) consisting of 132 K-gluconate, 4.4 Na-gluconate, 4.4 NaCl, 2.2 MgCl₂, 1.1 EGTA, 11 HEPES, 22 sucrose, 14 TRIS creatine phosphate, 4 Mg-ATP, 0.3 and TRIS-GTP. Stable whole-cell recordings were acquired at 100 kHz in pClamp 10.3 and pClamp 10.7 (Molecular Devices, San Jose, CA, USA) via an Axon 700B multiclamp amplifier and Digidata 1440 digitizer. Cells were visually identified and included for recording based on an access resistance below 20MΩ, and the demonstration of a robust fast inward sodium current in voltage clamp and overshooting action potentials in current clamp.

Nashef A., Spindle M.S., Calame D.J, & Person A.L. (2023). A dual Purkinje cell rate and synchrony code sculpts reach kinematics. bioRxiv.

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Voltage Clamp Recordings from Oocytes and Cells

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Two-electrode voltage clamp (TEVC) recordings from X. laevis oocytes were performed one to three days after cRNA injection using an OC-725C Oocyte Clamp amplifier (Warner Instruments, Hamden, CT, USA), a Digidata 1322A Series (Axon Instruments, Foster City, CA, USA) and pClamp 10 software (Molecular Devices, San José, CA, USA). Current recordings from CHO cells were carried out using the whole-cell patch clamp technique with an Axopatch 200B amplifier (Axon Instruments, Foster City, CA, USA), an Axon Digidata 1550B series (Axon Instruments, Foster City, CA, USA), and pClamp 10 software (Molecular Devices, San José, CA, USA). For detailed information about solutions and test protocols please refer to the supplementary methods.

Ratte A., Wiedmann F., Kraft M., Katus H.A, & Schmidt C. (2019). Antiarrhythmic Properties of Ranolazine: Inhibition of Atrial Fibrillation Associated TASK-1 Potassium Channels. Frontiers in Pharmacology, 10, 1367.

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Whole-cell Patch Clamp and MEA Recording

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Whole cell patch clamp recording was performed on DIV 21 cultures using an Axopatch 200B amplifier with a Digidata 1440A digital acquisition system and pClamp 10 software (Molecular Devices). Experiments were performed at 37°C in atmospheric air using an extracellular solution containing (in mM): 144 NaCl, 5.3 KCl, 2.5 CaCl2, 1 MgCl2, 10 HEPES, 10 mM glucose, pH 7.4. The pipette solution contained (in mM): 130 mM K + gluconate, 4 mM NaCl, 0.5 mM CaCl2, 10 mM HEPES, 0.5 EGTA pH 7.2. Borosilicate glass pipettes were pulled to a resistance of 3-8 MΩ. Liquid junction potentials were measured as 20 mV and traces were offset by this value. For microelectrode arrays, cultures were plated and maintained on commercial MEAs (60MEA200/30iR-Ti-gr; Multi Channel Systems, Reutlingen, Germany) as described previously (Bijland et al., 2019) (link). A fluorinated ethylene-propylene membrane (ALA MEA-MEM-SHEET) sealed the MEA culture dishes. The recordings were done in differentiation medium. Signals were digitally filtered at 3 Hz high pass filter, 1 kHz low pass filter and amplified up to x20,000. A digital notch filter was used to remove 60 Hz noise during recording. For data acquisition and analysis, spikes and potentials were sorted and counted from 3-minute gap-free recordings using the pCLAMP10 software (Molecular Devices Corporation, California, USA).

Berghoff S.A., Spieth L., Sun T., Hosang L., Depp C., Sasmita A.O., Vasileva M.H., Scholz P., Zhao Y., Krueger‐Burg D., Wichert S.P., Brown E.R., Michael K., Nave K., Bonn S., Odoardi F., Rossner M.J., Ischebeck T., Edgar J.M., & Saher G. (2021). Neuronal cholesterol synthesis is essential for repair of chronically demyelinated lesions in mice.

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Patch-Clamp Recording of Neuronal Properties

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Following incubation, slices were transferred to a recording chamber, where oxygenated ACSF was warmed to 32°C and continuously superfused over the submerged slice at 3.3 ml/min. Recording electrodes of 3–5 MΩ tip resistance were pulled from borosilicate glass capillary tubing (Sutter Instruments) using a Flaming-Brown puller (Sutter Instruments). Recordings were collected from the CeM with patch pipettes filled with experiment-specific internal solutions (see below). Membrane properties (averaged across entire recordings) were provided by the membrane test in pClamp 10 (Molecular Devices) and electrophysiology data were acquired with a MultiClamp 700B (Molecular Devices, Sunnyvale, CA) at 10 kHz, filtered at 1 kHz, and stored for later analysis using pClamp 10 software (Molecular Devices).

Godin J.G, & Dugatkin L.A. (1996). Female mating preference for bold males in the guppy, Poecilia reticulata.. Proceedings of the National Academy of Sciences of the United States of America, 93(19), 10262-10267.

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Cardiac Cluster Calcium Transient and Contraction Analysis

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For calcium transient analysis, cardiac clusters were dissociated into single cells and
stained with Fluo-4, AM (Thermo Fisher Scientific) for 15 min at 37°C. Using C-Pace EP
(IonOptix, MA, USA), cells were paced at either 0.25 or 0.5 Hz and calcium transients were
recorded using MetaMorph Imaging System (Molecular Devices, CA, USA). Calcium transients
were analysed using pCLAMP 10 (Molecular Devices). For contraction analysis, cardiac
clusters were paced at 0.25 Hz using C-Pace EP (IonOptix) and video recordings were
acquired using CKX41 Inverted Microscope (Olympus). The acquired videos were first
analysed using Musclemotion²⁰ (link)
and the raw data generated was then re-analysed using pCLAMP 10 (Molecular Devices).

Ramachandra C.J., Kp M.M., Chua J., Hernandez-Resendiz S., Liehn E.A., Knöll R., Gan L.M., Michaëlsson E., Jonsson M.K., Ryden-Markinhuhta K., Bhat R.V., Fritsche-Danielson R., Lin Y.H., Sadayappan S., Tang H.C., Wong P., Shim W, & Hausenloy D.J. (2021). Inhibiting cardiac myeloperoxidase alleviates the relaxation defect in hypertrophic cardiomyocytes. Cardiovascular Research, 118(2), 517-530.

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