Whole-cell patch-clamp recordings were performed from CA1 pyramidal cells visualised using infrared differential contrast imaging. Thin-walled borosilicate glass capillaries were used to fabricate recording electrodes with a resistance of 2.5–3.5 MΩ. Intracellular pipette solution contained (in mM) KCH3O3S 135, HEPES 10, Na2-Phosphocreatine or di-Tris-Phosphocreatine 10, MgCl2 4, Na2-ATP 4, Na-GTP 0.4, 5 QX-314-Bromide (pH adjusted to 7.2 using KOH, osmolarity 290–295). Cell-impermeable Ca2+ dyes detailed below and the Ca2+ insensitive morphological tracer Alexa Fluor 594 hydrazide (50 μM) were routinely added to the intracellular solution. Throughout recordings cells were held in voltage clamp at −65 mV, and the aCSF routinely supplemented with 100 µM Picrotoxin (Tocris Bioscience) and 30 µM D-Serine. Electrophysiological recordings were carried out by using a set of remotely controlled micromanipulators and XY-translation stage (Luigs and Neumann) with a Multiclamp 700B amplifier (Molecular Devices). Signals were digitised at 10 kHz and stored for off-line analysis using WinWCP V4.1.5–4.7 (John Dempster, University of Strathcyde) or PClamp 10.4–10.5 (Molecular Devices).
Pclamp 10 4 10
Manufactured by Molecular Devices
PClamp 10.4–10.5 is a software suite for electrophysiology data acquisition and analysis. It provides tools for recording and analyzing electrical signals from cells, tissues, and other biological preparations.
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2 protocols using pclamp 10 4 10
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Whole-cell Patch-clamp Recordings of CA1 Pyramidal Cells
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Patch-clamp recording of CA1 pyramidal cells
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Whole-cell patch-clamp recordings were performed from CA1 pyramidal cells visualised using infrared differential contrast imaging. Thin-walled borosilicate glass capillaries were used to fabricate recording electrodes with a resistance of 2.5-3.5 MΩ. Intracellular pipette solution contained (in mM) KCH3O3S 135, HEPES 10, Na2-Phosphocreatine or di-Tris-Phosphocreatine 10, MgCl2 4, Na2-ATP 4, Na-GTP 0.4, 5 QX-315-Bromide (pH adjusted to 7.2 using KOH, osmolarity 290-295). Cellimpermeable Ca 2+ dyes detailed below and the Ca 2+ insensitive morphological tracer Alexa Fluor 594 hydrazide (50 μM) were routinely added to the intracellular solution.
Throughout recordings cells were held in voltage clamp at -65mV, and the aCSF routinely supplemented with 100µM Picrotoxin (Tocris Bioscience) and 30µM D-Serine. Electrophysiological recordings were carried out by using a set of remotely controlled micromanipulators and XY-translation stage (Luigs and Neumann) with a Multiclamp 700B amplifier (Molecular Devices). Signals were digitized at 10 kHz and stored for off-line analysis using WinWCP V4.1.5 -4.7 (John Dempster, University of Strathcyde) or PClamp 10.4-10.5 (Molecular Devices)
Throughout recordings cells were held in voltage clamp at -65mV, and the aCSF routinely supplemented with 100µM Picrotoxin (Tocris Bioscience) and 30µM D-Serine. Electrophysiological recordings were carried out by using a set of remotely controlled micromanipulators and XY-translation stage (Luigs and Neumann) with a Multiclamp 700B amplifier (Molecular Devices). Signals were digitized at 10 kHz and stored for off-line analysis using WinWCP V4.1.5 -4.7 (John Dempster, University of Strathcyde) or PClamp 10.4-10.5 (Molecular Devices)
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