Tetrodes were lowered in steps of 60 μm before each day of recording. The neuronal activity and the onset of PP stimulation were simultaneously recorded with acquisition equipment (Open Ephys) via an RHD2132 amplifier board (Intan Technologies). Signals were monitored and recorded from 32 low-noise amplifier channels at 30 kHz, band-pass filtered (0.3–7.5 kHz). To achieve spike activity, the raw data were high-pass filtered at 300 Hz with subsequent thresholding and offline sorting by commercial software (Offline Sorter, Plexon). The threshold was lower than the 3-sigma peak heights line and optimized manually based on the signal-to-noise ratio. The features of three valley electrodes were used for spike sorting. Trials were aligned to the initiation of the peripheral stimulus to compute the peristimulus time histogram (PSTH) for each single unit using MATLAB (MathWorks).
Rhd2132 amplifier
Manufactured by Intan Technologies
Sourced in United States
The RHD2132 is a high-performance amplifier designed for laboratory applications. It features a low-noise input stage and high-bandwidth signal processing capabilities. The core function of this device is to amplify and condition electrical signals for further analysis and measurement.
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6 protocols using rhd2132 amplifier
1
Simultaneous Neural Recordings and Electrical Stimulation
2
Optogenetic Silencing in Freely Behaving Animals
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Animals were allowed to move freely in different environments to stimulate exploratory behavior (open field, small home cage with bedding, large home cage with floor covered by paper tissue, platform without lateral walls). Electrical signals were recorded at a sampling frequency of 30,000 Hz using an Intan RHD2132 amplifier (Intan Technologies, USA) and the OpenEphys acquisition software (https://open-ephys.org ). To induce optogenetic silencing, the recorded brain area was illuminated with green light of 561 nm wavelength emitted from a laser diode (LASOS Lasertechnik GmbH, Germany). Light intensity was adjusted to ~ 10 mW at the tip of the light guide.
3
Simultaneous LFP and Ca2+ Imaging Analysis
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Local LFP signals were recorded using the RHD2132 amplifier board (Intan Technologies, version 1.41) with an amplifier bandwidth from 0.1 Hz to 7.5 kHz and sampled at 20 kHz. Data were analyzed and SPW-Rs were then detected using MATLAB-based software (MES5 version 2043-MES6 version 11391, Femtonics). In order to preserve the phase and amplitude of individual ripple cycles in the LFP signal, we used the difference between two low-pass filters as previously described36 (link). To eliminate unit activity, we set the two cut-off frequencies of the two Gaussian filters to 150 Hz and 500 Hz. Only the channel with the highest ripple band LFP power, and events exceeding 4 SD over the baseline were analyzed. Signals were discarded when they were also present on the contralateral cortical electrode. The electrophysiological and the simultaneously recorded Ca2+ imaging data were overlaid and analyzed in a MATLAB-based program (MES, Femtonics) with a delay between ripple peak and Ca2+ peak of 29.02 ± 4.73 ms.
4
Simultaneous Neural Recordings and Electrical Stimulation
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Tetrodes were lowered in steps of 60 μm before each day of recording. The neuronal activity and the onset of PP stimulation were simultaneously recorded with acquisition equipment (Open Ephys) via an RHD2132 amplifier board (Intan Technologies). Signals were monitored and recorded from 32 low-noise amplifier channels at 30 kHz, band-pass filtered (0.3–7.5 kHz). To achieve spike activity, the raw data were high-pass filtered at 300 Hz with subsequent thresholding and offline sorting by commercial software (Offline Sorter, Plexon). The threshold was lower than the 3-sigma peak heights line and optimized manually based on the signal-to-noise ratio. The features of three valley electrodes were used for spike sorting. Trials were aligned to the initiation of the peripheral stimulus to compute the peristimulus time histogram (PSTH) for each single unit using MATLAB (MathWorks).
5
Optogenetic Manipulation of Sleep-Wake Circuits
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The SI Appendix includes detailed methods. DBH-Cre mice and TH-Cre mice were used. All of the procedures were approved by Institutional Animal Care and Use Committees of the University of Pennsylvania and were done in accordance with the federal regulations and guidelines on animal experimentation (National Institutes of Health Offices of Laboratory Animal Welfare Policy). For sleep recordings, EEG and EMG signals were recorded using an RHD2132 amplifier (Intan Technologies) connected to the RHD USB Interface Board (Intan Technologies). For fiber photometry, we used a Tucker-Davis Technologies RZ5P amplifier. For optogenetic stimulation, light pulses were generated by a blue laser (473nm, Laserglow) and sent through the optic fiber (ThorLabs) that connects to the ferrule on the mouse head. At the end of the experiment, mice were deeply anesthetized and transcardially perfused with phosphate-buffered saline (PBS) followed by 4% paraformaldehyde in PBS for subsequent histology to confirm virus expression and optic fiber placement.
6
Multichannel Neural Recording in Mouse Prefrontal Cortex
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Animals were anaesthetized with a mixture of ketamine/xylazine (75:1, 0.35 ml/28 g i.p.) and this state was maintained with an inhalation mask providing 1.5% isoflurane. Several microscrews were placed into the skull to stabilize the implant, and the one on top of the cerebellum was used as a general ground. The probe was implanted in the prefrontal cortex (coordinates: AP, 1.5 mm; ML, ±0.5 mm; DV, −1.7 mm from bregma). The implantation was performed by coating the probe with maltose (see protocol below) to provide temporary probe stiffness and facilitate probe insertion. The probe was sealed with dental cement. TDT-ZifClip connectors were used to connect the probe to the electrophysiological system via a miniaturized cable. After the surgery, the mouse underwent a recovery period of 1 week receiving analgesia (buprenorphine) and anti-inflammatory (meloxicam) treatments. Neural activity was recorded with the multichannel Open Ephys system at a sampling rate of 30 kHz with an Intan RHD2132 amplifier. The auditory task experiments were conducted in a soundproofed box, with two speakers inside using protocols based on previously described work61 (link). The sound stimulus consisted of a 15-ms-long white noise click, repeated 100 times (cycles), each separated by 5 s (interstimulus interval). During the task, the animal was able to move freely.
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