Axon patch 200b amplifier

Manufactured by Molecular Devices

Sourced in United States

The Axon-patch 200B amplifier is a versatile laboratory equipment designed for electrophysiology experiments. It is a high-performance patch-clamp amplifier that can be used to measure and record electrical signals from single cells or small tissue samples. The core function of the Axon-patch 200B is to amplify and condition the small electrical signals generated by the cells, allowing researchers to study the underlying cellular processes with high precision and sensitivity.

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Axon 200B amplifier (24 citations)

21 protocols using axon patch 200b amplifier

TRPM2 Channel-Mediated Current Recordings

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Patch-clamp recordings were made in the whole-cell configuration at room temperature using an Axonpatch 200B amplifier (Molecular Devices, Sunnyvale, CA, USA). Cells were kept in extracellular solution (ECS) containing (in mM): 147 NaCl, 2 KCl, 1 MgCl₂, 2 CaCl₂, 10 HEPES, and 13 glucose, pH 7.4. Electrodes had a final resistance of 3–5 MΩ when filled with intracellular solution containing (in mM): 147 NaCl, 0.05 EGTA, 1 MgCl₂, 10 HEPES, and 0.5 ADPR, pH 7.3. The membrane potential was held at 0 mV. To record ADPR-induced currents, voltage ramps with 500 ms duration from −100 mV to 100 mV were applied every 5 s. The inward currents at −80 mV, denoted by circles, are shown in the figures. To confirm that they were TRPM2 channel-mediated currents, we applied ECS at pH 5.0, which completely blocks TRPM2 channels36 (link), at the end of each measurement. Change of the acidic ECS was carried out using an RSC-160 system (Bio-logic Science Instruments, Claix, France). For analysis, the mean of the first three ramps before channel activation was used for leak subtraction of all subsequent current recordings.

Yu P., Li J., Jiang J., Zhao Z., Hui Z., Zhang J., Zheng Y., Ling D., Wang L., Jiang L.H., Luo J., Zhu X, & Yang W. (2015). A dual role of transient receptor potential melastatin 2 channel in cytotoxicity induced by silica nanoparticles. Scientific Reports, 5, 18171.

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Patch-Clamp Analysis of NMDA-Induced Currents

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NMDA-induced currents were recorded in transfected primary striatal neurons (DIV 11) at −60 mV by whole-cell patch clamping using an AxonPatch 200B amplifier (Molecular Devices, San Jose, CA, USA). The borosilicate glass micropipettes used had a resistance of 4–6 MΩ and were filled with the following internal solution (in mM): CsMeSO₄ 130, CsCl 10, CaCl₂ 0.5, EGTA 5, HEPES 10, and NaCl 10 (pH 7.3 adjusted with CsOH). Cells were perfused with extracellular solution containing 140 mM NaCl, 2.5 mM KCl, 1.8 mM CaCl₂, 10 mM HEPES, and 15 mM glucose supplemented with 10 μM glycine (pH 7.4 adjusted with NaOH). NMDA (100 μM) was diluted in the extracellular solution and rapidly perfused with a six-channel perfusion valve control system VC-77SP/perfusion fast-step SF-77B (Warner Instruments, Hamden, CT, USA). All experiments were performed at RT (22–25 °C). The currents were filtered at 1 kHz (4-pole low-pass Bessel filter) and digitized at a sampling rate of 10 kHz to a personal computer and analyzed with pClamp 10.7 software (Molecular Devices, San Jose, CA, USA).

Fão L., Coelho P., Rodrigues R.J, & Rego A.C. (2022). Restored Fyn Levels in Huntington’s Disease Contributes to Enhanced Synaptic GluN2B-Composed NMDA Receptors and CREB Activity. Cells, 11(19), 3063.

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Whole-Cell Patch-Clamp Electrophysiology

Cited 1 time

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Whole-cell patch-clamp recording was performed as previously described (Teng et al., 2019 (link)). Briefly, recordings were obtained with an Axonpatch 200B amplifier, digitized with a Digidata 1322a 16-bit data acquisition system, acquired with pClamp 8.2, and analyzed with Clampfit 9 or 10 (Molecular Devices). Records were sampled at 5 kHz and filtered at 1 kHz. Patch pipettes with a resistance of 2–4 MΩ were fabricated from borosilicate glass (Sutter Instrument). Solution exchange was achieved with a fast-step perfusion system (Warner Instrument, SF-77B) custom modified to hold seven microcapillary tubes in a linear array. Cells were treated with trypsin-EGTA and plated into the recording chamber immediately before each experiment. After a gigaohm seal was formed and whole-cell recording was achieved, the cell was lifted and moved in front of the microcapillary tubes. The membrane potentials were held at –80 mV unless otherwise indicated.

Teng B., Kaplan J.P., Liang Z., Chyung K.S., Goldschen-Ohm M.P, & Liman E.R. (2023). Zinc activation of OTOP proton channels identifies structural elements of the gating apparatus. eLife, 12, e85317.

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Patch-Clamping of MMSC-Derived Cardiomyocytes

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Patch-clamping was performed as previously described [32 (link), 33 (link)]. MMSC-derived cells grown on coverslips were placed on the stage of the electrophysiological recording setup and perfused with a bath solution that contained 140mM NaCl, 5.4mM KCl, 1mM MgCl₂, 1.8mM CaCl₂, 5mM Na-HEPES, 0.33mM NaH₂PO₄, 10mM glucose, and 10mM HEPES (pH7.4 NaOH). For whole-cell patch-clamp recordings from a single spontaneously contracting MMSC-derived cell, electrode pipettes were made from borosilicate glass capillaries by a P-97 puller (WPI instruments) and had resistances of 3-6 MΩ when filled with internal solution. The pipette internal solution contained 140mM KCl, 1mM MgCl₂, 10mM HEPES, 10mM EGTA, 5mM MgATP, 10mM TEA-Cl (pH7.3 KOH). Electrophysiological signals were recorded using an Axonpatch 200B amplifier, filtered at 2 kHz, and sampled at 5 kHz using pClamp9.0 software (Molecular Device). To test membrane excitability and action potential firing, cells were recorded under current-clamp configuration. Current steps were applied to cells from −100 to 300 pA with each step at 20 pA. Step currents were injected for a duration of 2 seconds for each step and the interval between each step was 2 seconds. For measurement of I_f, cells were held at −60 mV and hyperpolarized to −140 mV for 6 seconds with each step at 10 mV, and then brought to the holding potential of −60 mV.

Huang W., Liang J., Feng Y., Jia Z., Jiang L., Cai W., Paul C., Gu J.G., Stambrook P.J., Millard R.W., Zhu X.L., Zhu P, & Wang Y. (2018). Heterogeneity of adult masseter muscle satellite cells with cardiomyocyte differentiation potential. Experimental cell research, 371(1), 20-30.

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Measuring Tonic and Evoked Currents in Electrophysiology Recordings

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Recordings were obtained using an Axon-patch 200B amplifier (Molecular Devices, San Jose, CA, USA), low-pass filtered at 5 kHz (Bessel, 8-pole) and digitized at 10 kHz with a National Instruments data acquisition board (BNC 2110, National Instruments, Austin, TX, USA). All data were acquired with EVAN (custom-designed LabView-based software).
Tonic current measurement. A custom written procedure (Wavemetrics, IGOR Pro 6.22A, Lake Oswego, OR, USA) was used to perform the analysis. An all-points histogram of a randomly selected recording segment of 10 s during the period of interest was plotted. A Gaussian was fitted to the part of the distribution from the minimum value at the left to the rightmost (largest) value of the histogram distribution. The mean of the fitted Gaussian was considered to be the tonic current (I_tonic). This process was repeated for all segments of interest.
Evoked current measurement. Evoked currents were extracted from continuous current recording and baseline corrected. The measurement of amplitude and decay time is described in legend of Fig. S2.

Wei X., Nishi T., Kondou S., Kimura H, & Mody I. (2018). Preferential enhancement of GluN2B-containing native NMDA receptors by the endogenous modulator 24S-hydroxycholesterol in hippocampal neurons. Neuropharmacology, 148, 11-20.

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Electrophysiology Data Acquisition

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Data were digitized at 10kHz, and low-pass filtered at 2kHz using CED Micro 1401-4 A-D acquisition unit and Axon patch 200B amplifier (Molecular devices). All recordings were obtained using CED Signal 5 software.

Nair J.D., Wilkinson K.A., Yucel B.P., Mulle C., Vissel B., Mellor J, & Henley J.M. (2023). GluK2 Q/R editing regulates kainate receptor signaling and long-term potentiation of AMPA receptors. iScience, 26(10), 107708.

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Neonatal Mouse Diaphragm Neuromuscular Function

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Recording was performed as described previously (Li et al., 2008 (link); Wu et al., 2012a (link), 2012b (link); Barik et al., 2014a (link)). Neonatal mice diaphragms with ribs and intact phrenic nerves were dissected in oxygenated (95% O₂/5% CO₂) Ringer's solution (136.8 mM NaCl, 5 mM KCl, 12 mM NaHCO₃, 1 mM NaH₂PO₄, 1 mM MgCl₂, 2 mM CaCl₂, and 11 mM D-glucose, pH 7.3) and pinned on Sylgard gel in a dish perfused with oxygenated Ringer's solution. To measure mEPP, microelectrodes, 20–50 MΩ when filled with 3 M KCl, were pierced into the center of muscle fibers with the resting potential between −45 and −55 mV. To evoke end-plate potentials, phrenic nerves were stimulated by a suction electrode with suprathreshold square pulses (0.1 ms) using Master-8 (A.M.P.I, Jerusalem, Israel). Data were collected with axonpatch 200B amplifier (Molecular Devices, Sunnyvale, CA), digitized with Digidata 1322A (Molecular Devices, Sunnyvale, CA), and analyzed using pClamp 9.2 (Molecular Devices, Sunnyvale, CA).

Wu H., Barik A., Lu Y., Shen C., Bowman A., Li L., Sathyamurthy A., Lin T.W., Xiong W.C, & Mei L. (2015). Slit2 as a β-catenin/Ctnnb1-dependent retrograde signal for presynaptic differentiation. eLife, 4, e07266.

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Electrophysiology of Ion Channels

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Current was recorded with an Axon‐patch 200B amplifier (Molecular Devices, CA, USA) and analysed with ClampFit 10.2 software (Molecular Devices). I_Kv11.1, I_KIR2.1, I_Nav1.5, and I_Ca‐L were measured under room temperature (22°C). Action potential and I_K were measured at 37°C.

Qile M., Beekman H.D., Sprenkeler D.J., Houtman M.J., van Ham W.B., Stary‐Weinzinger A., Beyl S., Hering S., van den Berg D., de Lange E.C., Heitman L.H., IJzerman A.P., Vos M.A, & van der Heyden M.A. (2019). LUF7244, an allosteric modulator/activator of Kv11.1 channels, counteracts dofetilide‐induced torsades de pointes arrhythmia in the chronic atrioventricular block dog model. British Journal of Pharmacology, 176(19), 3871-3885.

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Neuronal Excitability Measurement Protocol

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Current-clamp recordings to measure neuronal excitability were performed with a modified protocol as previously described21 (link)33 (link)34 (link). Electrophysiology recordings were performed by experimenters who were blind to genotype. All recordings were performed at 31 ± 1°C. Neuronal intrinsic excitability was measured as the number of spikes in response to a series of fixed 500 ms current injection steps (0, 0.03, 0.06, 0.09, 0.12, 0.15, 0.18, and 0.21 nA). Data were acquired by Axonpatch 200B amplifier at 2 kHz with pClamp 9.2 software (Molecular Devices, Sunnyvale, CA) and analyzed with clampfit 9.2 software (Molecular Devices, Sunnyvale, CA) and Mini Analysis software (Synaptosoft, Decatur, GA). For field potential recordings from slices of hippocampal DG, a stimulating electrode was placed in the angular bundle of the perforant path and a recording electrode in the hilus of the DG. A single slice was transferred to a liquid-air interface chamber (Fine Science Tools, Inc., Foster City, CA, USA) and suspended on a nylon net at the liquid-air interface in a bath of continuously dripping oxygenated ACSF (2–2.5 ml/min).

Liu Q., Xin W., He P., Turner D., Yin J., Gan Y., Shi F.D, & Wu J. (2014). Interleukin-17 inhibits Adult Hippocampal Neurogenesis. Scientific Reports, 4, 7554.

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Electrophysiological Assessment of Spinal Cord

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Spinal slices obtained from the rats 14 days after surgery were used for electrophysiological assessments. All the rats were tested by mechanical threshold testing and signed with numbers before recording. Researchers were blinded to the rats. Under inhaled isoflurane anesthesia, the L3‐L5 spinal cord was passively separated from the vertebrae and quickly immersed in ice‐cold oxygenated high‐sucrose ACSF containing (in mM): 234 sucrose, 3.6 mM KCl, 1.2 mM MgCl₂, 1.2 mM NaH₂PO₄, 12 mM glucose, 2.5 mM CaCl₂, and 25 mM NaHCO₃ for 90 seconds. The L3‐L5 spinal cord was mildly blew with 50 mL syringe and immersed in ice‐cold ACSF. The spinal cords were covered by 2% agarose, and the 400‐μm spinal slices were cut on a vibratome (Leica VT‐1200S, Wetzlar, Germany), and more importantly, forceps were used to sign the other sides to distinguish the operated sides. The spinal slices were transferred and incubated in an oxygenated artificial cerebrospinal fluid (ACSF: 125 mM NaCl, 3 mM KCl, 1.25 mM NaH₂PO₄, 26 mM NaHCO₃, 1 mM MgCl₂, 2 mM CaCl₂, and 10 mM D‐glucose, pH 7.3) for 30 minutes at 32℃ and cooling to room temperature for one hour, and then transferred to the recording chamber. Data acquisition was conducted by using an Axonpatch 200 B amplifier (Axon Instruments), and data were filtered at 2 kHz and digitized at 5 kHz using pClamp10 software.⁴³ (link)

Ma L., Peng S., Wei J., Zhao M., Ahmad K.A., Chen J, & Wang Y. (2021). Spinal microglial β‐endorphin signaling mediates IL‐10 and exenatide‐induced inhibition of synaptic plasticity in neuropathic pain. CNS Neuroscience & Therapeutics, 27(10), 1157-1172.

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