For whole-cell recordings, the extracellular solution contained 140 mM NaCl, 6 mM KCl, 1 mM MgCl2, 10 mM glucose, and 10 mM HEPES (pH 7.4). Recording electrode pipettes were made of borosilicate glass (Sutter Instruments) and fire-polished to a resistance between 2.5 and 4 MΩ. Pipettes were filled with an intracellular solution consisting of 140 mM CsCl, 5 mM EGTA, 1 mM MgCl2, and 10 mM HEPES (pH 7.2). Cells were recorded under voltage-clamp conditions using an Axopatch 200A (Molecular Devices, Union City, CA, USA) using 1s-ramps from −80 to +80 mV, delivered once per second, and a sampling rate of 10 kHz. Currents were analyzed offline using Clampfit v10.4 (Molecular Devices) and plotted using OriginLab software.
Clampfit v10
Manufactured by Molecular Devices
Sourced in United States
Clampfit v10.4 is a data analysis software designed for electrophysiology experiments. It provides tools for the analysis and visualization of electrophysiological data, including voltage-clamp and current-clamp recordings.
Automatically generated - may contain errors
Lab products found in correlation
Axopatch 200a, by Molecular Devices (2 mentions)
Borosilicate glass, by Sutter Instruments (2 mentions)
Prism v9, by GraphPad (2 mentions)
Originpro v 9, by OriginLab (2 mentions)
Originpro v 8, by Molecular Devices (2 mentions)
Matlab, by MathWorks (1 mentions)
Originpro 2016, by OriginLab (1 mentions)
Datacontroller384 v1, by Nanion Technologies (1 mentions)
Sigmaplot 2000, by Grafiti LLC (1 mentions)
Labchart, by ADInstruments (1 mentions)
Prism 9, by GraphPad (1 mentions)
Premiere pro cc 2018, by Adobe (1 mentions)
Matlab r2015a, by MathWorks (1 mentions)
Spss statistics 23, by IBM (1 mentions)
Cgp54626, by Bio-Techne (1 mentions)
Snap 5114, by Bio-Techne (1 mentions)
No 711, by Bio-Techne (1 mentions)
Matlab v 2018a, by MathWorks (1 mentions)
Statistica 8, by StatSoft (1 mentions)
26 protocols using clampfit v10
1
Whole-Cell Voltage-Clamp Recordings
2
Quantifying Synaptic Inhibition Dynamics
3
Sodium Current Analysis Protocol
4
Whole-Cell Voltage-Clamp Recordings
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For whole-cell recordings, the extracellular solution contained 140 mM NaCl, 6 mM KCl, 1 mM MgCl2, 10 mM glucose, and 10 mM HEPES (pH 7.4). Recording electrode pipettes were made of borosilicate glass (Sutter Instruments) and fire-polished to a resistance between 2.5 and 4 MΩ. Pipettes were filled with an intracellular solution consisting of 140 mM CsCl, 5 mM EGTA, 1 mM MgCl2, and 10 mM HEPES (pH 7.2). Cells were recorded under voltage-clamp conditions using an Axopatch 200A (Molecular Devices, Union City, CA, USA) using 1s-ramps from −80 to +80 mV, delivered once per second, and a sampling rate of 10 kHz. Currents were analyzed offline using Clampfit v10.4 (Molecular Devices) and plotted using OriginLab software.
5
Neuronal Electrophysiology and Receptor Binding
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Results are presented as mean ± standard deviation (S.D.) or mean with 95% confidence intervals (C.I.). Pairwise comparisons of data in the absence and presence of XOB were made using an unpaired t-test (Table 4) or paired t-test (Table 6) with P < 0.05 considered significant. For automated patch clamp comparisons (Table 5), mixedeffects analysis was performed with Bonferroni multiple comparisons and adjusted P value < 0.05 considered significant. Experiments in this exploratory study were designed to test biological hypotheses and calculated P-values should be interpreted as descriptive. Manual I Na data was collected using Clampfit v10.4 (Molecular Devices), and automated I Na data was quality controlled and exported using Nanion DataControl384 software. All data analysis of recorded I Na and 5-HT receptor binding was performed using the software packages Microsoft Excel and Graph Pad Prism.
Active and passive neuronal properties of brain slice recordings were analyzed using custom MATLAB (MathWorks) software.
Active and passive neuronal properties of brain slice recordings were analyzed using custom MATLAB (MathWorks) software.
6
Quantitative Analysis of KCNQ1-KCNE1 Channel Kinetics
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Data were collected for each experimental condition from at least three transfections and analyzed and plotted using DataController384 V1.2.1 (Nanion Technologies, Munich, Germany), Clampfit V10.4 (Molecular Devices Corp.), Excel (Microsoft Office 2013, Microsoft), SigmaPlot 2000 (Systat Software, Inc, San Jose, CA, USA) and OriginPro 2016 (OriginLab, Northampton, MA, USA) software. Whole-cell currents were normalized for membrane capacitance and results expressed as mean ± SEM. The number of cells used for each experimental condition and the threshold for statistical significance (p<0.001) are given in the figure legends or table footnotes. Additional custom semi-automated data handling routines were used for rapid analysis of current density, voltage-dependence of activation, and gating kinetics. The voltage-dependence of activation was determined only for cells with mean current density greater than the background current amplitude. Since normalized KCNQ1-KCNE1 tail currents do not saturate at the potentials tested nor would they saturate at more depolarized potentials (see Figure 3C in Wang et al., 2020 (link) and Figure 3C in Wang et al., 2012 (link)), we refer to the V1/2 value determined by curve fitting as the ‘apparent’ activation V1/2 (V1/2app).
7
Analyzing Neurochemical Responses in Vitro
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Data were acquired using Clampfit—V10.4, Molecular Devices (San Jose, CA, USA) and data sheets were constructed in Excel (Microsoft) and GraphPad Prism Software. Two-way ANOVA were performed to compare the overall significance of the difference between OT and OT + Mg2+. Sidak’s multiple comparison test was used to determine differences at individual concentrations. Significance was set at p < 0.05.
8
Hippocampal Slice LTP and mEPSC Recording
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Tetanic stimulation-evoked fEPSP in CA1 neurons of hippocampal slices was recorded following the protocols previously described (Chao et al., 2013) . Briefly, baseline responses were recorded for at least 20 min under 0.017 Hz, 0.1-ms pulse duration, and biphasic stimulations. LTP was evoked by 100-Hz HFS or TBS (9 bursts of 4 pulses at 100 Hz, 200-ms inter-burst interval). For 4 trains of stimulations, the inter-train intervals were 5 min. Data were normalized to the 20-min baseline. The mEPSC recordings were performed with modifications (Chen and Hsueh, 2012) . The extracellular solution contained (in millimolars) 136.5 NaCl, 5.4 KCl, 1.8 CaCl 2 , 0.53 MgCl 2 , 5.56 D-glucose, 5 HEPES (pH 7.4), 0.001 tetrodotoxin, and 0.02 bicuculline. The intracellular solution contained (in millimolars) 140 K-gluconate, 5 NaCl, 2 EGTA, 10 HEPES (pH 7.3), 4 MgATP, and 0.3 NaGTP. Neurons cultured on coverslips at DIV14-21 were whole-cell voltage-clamped at À70 mV to record mEPSCs, and data were analyzed by using Clampfit (v.10.4) (Molecular Devices).
9
Spectral Analysis of Baseline EEG
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Analysis software packages Clampfit v10 (Molecular Devices) and SciWorks v10 (Datawave Technologies) were used. Spectral analysis of baseline EEG oscillations was performed with the fast Fourier transformation (FFT, 0.5 Hz resolution). The power of EEG activities was analyzed in three frequency bands: delta (1–4 Hz), sigma (10–17 Hz, spindles), and gamma (30–80 Hz). For each band, the total power was the sum of all FFT values. Power measures were averaged into 2 min blocks (60 values ± SEM) given as a percentage of change from the averaged values under the control condition (about 100%). Tested parameters and number of rats used for each condition are presented in Table S1. Statistical analyses were performed using the software R v3.6.1 (R Core Team, Vienna Austria, 2019). Comparison between vehicle condition (tDCS 0 mA) and tDCS condition was done using parametric tests: paired student’s t-test and one-way analysis of variance with a Tukey’s posthoc test HSD (“honestly significant difference,” significance level p < 0.05). The Wilcoxon test was used where data were not normally distributed. Each animal was its own control.
10
EEG Spectral Analysis Protocol
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Analysis software packages Clampfit v10 (Molecular Devices) and SciWorks v10 (Datawave Technologies) were used. Spectral analysis of baseline EEG oscillations was performed with the fast Fourier transformation (FFT, 0.5 Hz resolution). The power of EEG activities was analyzed in 3 frequency bands: delta-(1-4 Hz), sigma-(10-17 Hz, spindles), gamma-(30-80 Hz) frequency oscillations. For each band, the total power was the sum of all FFT values. Statistical analyzes were performed using the software R. Parametric tests were used to assess the significance of the results: paired student t test, oneway analysis of variance with a Tukey's post-hoc test HSD ("honestly significant difference", significance level p <0.05). Each animal was its own control.
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