The SG was easily discernible with transmitted illumination as a relatively translucent band across the dorsal horn in the transverse slice preparations (Fig. 1 ). Blind whole-cell voltage-clamp recordings were made from SG neurons, as previously described [17 (link),19 (link)]. The patch pipettes were filled with a solution containing potassium gluconate solution (in mM): K-gluconate 135, KCl 5, CaCl2 0.5, MgCl2 2, EGTA 5, HEPES 5, and ATPMg 5 (pH 7.2). The tip resistance of the patch pipettes was 6–12 MΩ. Series resistance was assessed according to the response to a 5-mV hyperpolarizing step. This value was monitored during the recording session, and data were rejected if values changed by >15%. Signals were acquired with a patch-clamp amplifier (Axopatch 700A, Molecular Devices, Union City, CA, USA). The data were digitized with an AD/DA converter (Digidata 1321A, Molecular Devices), stored on a personal computer using a data acquisition program (Clampex, version 9.0, Molecular Devices), and analyzed using a software package (Clampfit, version 9.0, Molecular Devices) (Fig. 1 ). Cell recordings were obtained in voltage-clamp mode at holding potentials of -70 mV to record EPSCs [17 (link),19 (link)].
Digidata 1321a
Manufactured by Molecular Devices
Sourced in United States
The Digidata 1321A is a high-performance data acquisition system designed for electrophysiology research. It features 16-bit analog-to-digital conversion with a sampling rate of up to 200 kHz per channel. The Digidata 1321A provides reliable and precise data acquisition capabilities for a wide range of electrophysiological applications.
Automatically generated - may contain errors
Lab products found in correlation
Axopatch 700a, by Molecular Devices (2 mentions)
Clampfit, by Molecular Devices (1 mentions)
Clampex, by Molecular Devices (1 mentions)
Axoclamp 700b amplifier, by Molecular Devices (1 mentions)
Pclamp 8 acquisition software, by Molecular Devices (1 mentions)
Axopatch 1d amplifier, by Molecular Devices (1 mentions)
5 protocols using digidata 1321a
1
Whole-cell voltage-clamp recordings of substantia gelatinosa neurons
2
Patch-clamp recordings of spinal dorsal horn neurons
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The SG was easily discernible with transmitted illumination as a relatively translucent band across the dorsal horn in the transverse slice preparations (Fig. 1A ). Blind whole-cell voltage clamp recordings were made from SG neurons, as previously described [14 (link),15 (link),19 (link),20 (link)]. The patch pipettes were filled with a solution containing potassium gluconate solution (in mM): K-gluconate 135, KCl 5, CaCl2 0.5, MgCl2 2, EGTA 5, HEPES 5, and ATP-Mg 5 (pH, 7.2). The tip resistance of the patch pipettes was 6–12 MΩ. Series resistance was assessed according to the response to a 5-mV hyperpolarizing step. This value was monitored during the recording session, and data were rejected if values changed by >15%. Signals were acquired with a patch clamp amplifier (Axopatch 700A, Molecular Devices, Union City, CA, USA). The data were digitized with an analog to digital/digital to analog converter (Digidata 1321A, Molecular Devices), stored on a personal computer using a data acquisition program (Clampex version 9.0, Molecular Devices), and analyzed using a software package (Clampfit version 9.0, Molecular Devices). Cell recordings were made in voltage-clamp mode at holding potentials of -70 mV to record EPSCs [14 (link),15 (link),19 (link)].
3
Electrophysiology of Larval NMJ in Drosophila
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Current clamp recordings using sharp electrodes [using pClamp 10, an Axoclamp 700B amplifier and Digidata 1321A (Molecular Devices, USA)] were made from ventral longitudinal muscle 6 in abdominal segments 2 and 3 of third instar wandering larvae in haemolymph-like (HL-3) solution containing 1.5 mm Ca2+ as described previously (70 (link)). Recording electrodes (10–20 MΩ) were filled with 3 M KCl. All eEJP/mEJPs were recorded from muscles with resting potentials more negative than −60 mV and at 22°C as differences in recording temperature cause changes in glutamate receptor kinetics and amplitudes (71 (link)). All eEJP amplitudes were corrected for non-linear summation (72 (link)). Larval rearing temperatures were 25°C to avoid temperature-dependent fluctuations of NMJ morphology and evoked responses (73 (link)). QC was calculated by dividing the mean eEJP amplitudes by the mean mEJP amplitude of a given cell. mEJPs and eEJPs were low-pass filtered at 1 kHz. Data were collected from averaged multiple eEJPs and 60 s of mEJP recordings per muscle.
4
Extracellular Field Potential Recordings in Organotypic Neocortical Slices
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Organotypic neocortical slices were placed in a conventional submerged chamber and continuously perfused with oxygenated artificial cerebrospinal fluid (aCSF) (95% O2–5% CO2) at 32–34°C with a flow rate of 5–8 mL/min. aCSF contained the following compounds (mM): NaCl (120), KCl (3.3), CaCl2 (1.3), MgCl2 (1.3), NaH2PO4 (1.25), NaHCO3 (25), and d ‐glucose (11) with pH 7.3–7.4 when bubbled with 95% O2 and 5% CO2. Extracellular field potential recordings in neocortical layer II/III were performed using tungsten microelectrodes and a low‐noise multichannel amplifier (Dagan EX 4‐400) with a 1000 gain and digitized at 2 kHz using an analog to digital converter DigiData 1321A (Molecular Devices, Sunnyvale, CA.).
5
Patch-Clamp Analysis of mPanx1Δ371
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Whole cell currents were obtained with 3.0~4.5 MΩ borosilicate glass pipettes from Neuro2a cells transiently transfected with mPanx1Δ371. Cells were continuously perfused (~3 mL/min) at room temperature (23~25 °C) with a bath solution (pH 7.4) containing (in mM) 140 NaCl, 5 KCl, 2 CaCl2, 1 MgCl2, 10 glucose and 10 HEPES. To substitute extracellular Cl− with gluconate−, NaCl was replaced with equimolar Na-gluconate. The intracellular solution contained 140 CsCl, 5 NaCl, 5 MgCl2, 5 EGTA and 10 HEPES (pH 7.4). mPanx1Δ371-expressing cells were identified by GFP fluorescence. Whole-cell currents were recorded with an Axopatch 1D amplifier and a Digidata 1321A interface using pCLAMP 8 acquisition software (Molecular Devices, Sunnyvale, CA, USA). The current data were low-pass filtered at 500 Hz with a four-pole Bessel filter and digitized at 2 kHz. Cells were held at −100 mV and ramp voltage pulses (−100 mV to +100 mV over 500 ms; +100 mV for 125 ms) were applied at 5 s intervals.
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