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Axograph 4

Manufactured by Molecular Devices
Sourced in United States, United Kingdom

Axograph 4.6 is a software application designed for data analysis and visualization. It provides a range of tools for the processing and analysis of experimental data, including electrophysiological recordings, waveforms, and time-series data.

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5 protocols using axograph 4

1

Extracellular and Intracellular Recordings from Prefrontal Cortex Neurons

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Single extracellular field and intracellular recordings were made using a standard interface recording chamber. Extracellular recording electrodes were filled with normal ACSF (resistance 2–5 MΩ). Intracellular recording electrodes were made using 2 m potassium acetate-filled glass microelectrodes pulled to resistances of between 70 and 120 MΩ. Intracellular recordings from PFC neurons were performed in normal ACSF or in the presence of network oscillations. Only data recorded from cells with resting membrane potential of at least −50 mV and spikes exceeding +55 mV were used. IPSPs and excitatory postsynaptic potentials (EPSPs) were recorded at holding potentials of −30 mV and −70 mV, respectively.
Data were recorded with an Axoclamp-2B amplifier (Axon Instruments Inc., Union City, CA, USA). Extracellular data were filtered at 0.001–0.4 kHz and intracellular signals were low pass filtered at 2 kHz using Neurolog filters (Digitimer, Welwyn Garden City, Herts, UK). Mains noise was subtracted from the signal with a Humbug (Digitimer). Data were re-digitised at 10 kHz using an ITC-16 interface (Digitimer). Data were recorded using Axograph 4.6 software (Axon Instruments Inc.) and saved for further analysis.
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2

Intracellular and Extracellular Recordings of Gamma Oscillations in CA3

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Extracellular recording electrodes were filled with normal ACSF (resistance 2–5 MΩ). Gamma frequency oscillations were evoked using the glutamatergic agonist kainate (KA) and field traces were recorded from the border between stratum radiatum and stratum lacunosum moleculare in CA3. Intracellular recordings used potassium acetate (2 M) filled glass microelectrodes (resistance 80 MΩ - 150 MΩ). Data were recorded with an Axoclamp-2B amplifier (Axon Instruments Inc., Union City, CA, USA). Extracellular data were filtered at 0.001–0.4 kHz and intracellular signals were low pass filtered at 2 kHz using Bessel filters. Mains noise was subtracted from the signal with a Humbug (Digitimer, Welwyn Garden City, Herts, UK). Data were redigitised at 10 kHz using an ITC-16 interface (Digitimer, Welwyn Garden City, UK). Data were recorded and analysed using Axograph 4.6 software (Axon Instruments Inc., Union City, CA, USA). Further offline analysis was performed using MATLAB (Mathworks, Natick, USA).
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3

Extracellular recording of gamma oscillations

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Extracellular recording electrodes were filled with normal ACSF (resistance 2-5 MW).
Gamma frequency oscillations were evoked using the cholinergic agonist carbachol (10 µM) and field traces were recorded from the border between stratum radiatum and stratum lacunosum moleculare in CA3. Data were recorded with an Axoclamp-2B amplifier (Axon Instruments Inc., Union City, CA, USA). Extracellular data were filtered at 0.001-0.4 kHz and intracellular signals were low pass filtered at 2 kHz using Bessel filters. Mains noise was subtracted from the signal with a Humbug (Digitimer, Welwyn Garden City, Herts, UK). Data were redigitised at 10 kHz using an ITC-16 interface (Digitimer, Welwyn Garden City, UK).
Data were recorded and analysed using Axograph 4.6 software (Axon Instruments Inc., Union City, CA, USA). Further offline analysis was performed using MATLAB (Mathworks, Natick, USA).
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4

Electrophysiological Analysis of Synaptic Responses

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The data were analyzed off-line with AxoGraph 4.6 software program (Molecular Devices). For all protocols, measurements were made of the fEPSP slope and PS amplitude. The fEPSP slope was determined on the rising phase of the fEPSP over a range (typically 40–60%) that allowed reliable measurements without interference from the presynaptic fiber volley (PSV) or the reflection from the somatic population spike, which occurs on the falling phase of the fEPSP. The amplitude of the PSV and PS were measured as the peak amplitude of the negative deflection that reflects that response from a baseline value. Baseline value for the PS was estimated from a tangent line fitted across the two positive peaks of the somatic synaptic response.
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5

Measuring Deactivation and Desensitization of GluA1

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HEK-293 FT cells were transfected with GluA1 iQ alone, or in combinations with IgSF11 at equal amounts, via calcium-phosphate method. Two days after transfection, to measure deactivation, outside-out membrane patches were excised and positioned in front of an ultrafast perfusion system mounted on a piezoelectric translator (Physic instruments). To determine the desensitization time constant, 10 mM glutamate was applied for 1 sec. Recovery from desensitization was measured using a paired pulse protocol and an increasing interpulse interval. Data analysis was performed using axograph 4.6 (Molecular devices) and graphed using Kaleidagraph 3.5 (Synergy software) and Excel 2004 (Microsoft). Statistical analysis was done using Instat 2.03 (Synergy software).
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