Graphene electrode fibers were coupled to a customized adapter and connected to a 20X amplifier coupled to an Omniplex data acquisition system (Plexon Inc. Dallas TX). The electrophysiological activity was recorded at 40KHz. A reference electrode consisted of a graphene fiber placed between the sternomastoid muscle and the skin or under the abdominal skin tissue for the VN or SN branches respectively. Confirmation of the neural nature of the recorded activity was done by reduction of the signal after topical lidocaine application on the nerves, and elimination of the signal after euthanasia. Off-line sorter software (Plexon Inc, v3.3.5) and NeuroExplorer software (Nex Technologies, v 4.135) were used for data analysis.
Omniplex data acquisition system
Manufactured by Plexon
Sourced in United States
The Omniplex data acquisition system is a versatile hardware and software platform designed for recording and analyzing neural signals. It provides the capability to simultaneously capture data from multiple channels, allowing researchers to study neural activity across various experimental settings.
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10 protocols using omniplex data acquisition system
1
Graphene-based Electrophysiological Recording
2
Primate Visual Cognition Experiments
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The experiments were conducted in the same laboratory as that described for the monkey portions of ref.8 (link). Briefly, the monkeys were seated in a darkened booth ~ 72 cm from a calibrated and linearized CRT display spanning ~ 31° horizontally and ~ 23° vertically. Data acquisition and stimulus control were managed by a modified version of PLDAPS69 (link), interfacing with the Psychophysics Toolbox70 (link)–72 and an OmniPlex data acquisition system (Plexon, Inc.).
The monkeys were prepared for behavioral training and electrophysiological recordings earlier73 (link),74 (link). Specifically, each monkey was implanted with a head-holder and scleral search coil in one eye73 (link). The search coil allowed tracking eye movements using the magnetic induction technique75 (link),76 (link), and the head-holder comfortably stabilized head position during the experiments. The monkeys also each had a recording chamber centered on the midline and tilted 38° posterior of vertical, allowing access to both the right and left SC.
The monkeys were prepared for behavioral training and electrophysiological recordings earlier73 (link),74 (link). Specifically, each monkey was implanted with a head-holder and scleral search coil in one eye73 (link). The search coil allowed tracking eye movements using the magnetic induction technique75 (link),76 (link), and the head-holder comfortably stabilized head position during the experiments. The monkeys also each had a recording chamber centered on the midline and tilted 38° posterior of vertical, allowing access to both the right and left SC.
3
Neuronal Activity Recording on LCE MEAs
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Three polydomain and planar-aligned LCE MEAs were sterilized by ethylene oxide for 12 h, and then de-gassed at 37 °C for 48 h. LCE MEAs were treated as described in Section 2.3 . Every other day, wide band extracellular potentials generated by cultured neurons were recorded for 5 min from 59 channels simultaneously at a 40 kHz sampling rate using an Omniplex data acquisition system (Plexon Inc., Dallas, TX, USA). Wideband data were band pass filtered (250–7000 Hz) and spikes were detected by voltage excursions exceeding a threshold set to 5.5σ based on RMS noise on a per electrode basis. Spikes were manually sorted using Plexon’s offline sorter (Plexon Inc., Dallas, TX, USA). Additional data and statistical analyses were carried out using OriginPro software (Origin Labs, Farmington, ME, USA). Average spike rates were calculated using Neuroexplorer (NEX technologies, Reston, VA, USA). SNR was calculated as
where and are the mean peak-to-peak amplitude of the sorted unit and the RMS noise, respectively [43 (link)]. Active electrode yield percentage was calculated excluding electrodes with impedances over 5 MΩ that would not be capable of recording single units [5 (link)]. As a result, 8.2% of total electrodes were excluded.
where and are the mean peak-to-peak amplitude of the sorted unit and the RMS noise, respectively [43 (link)]. Active electrode yield percentage was calculated excluding electrodes with impedances over 5 MΩ that would not be capable of recording single units [5 (link)]. As a result, 8.2% of total electrodes were excluded.
4
Neural Signal Acquisition for Behavioral Study
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Neural recordings were digitized at the headstage (HST/32D, Plexon; Dallas, TX), passed through a passive commutator and collected using an Omniplex data acquisition system (Plexon). Signals were sampled at 40 kHz, bandpass filtered 0.10 Hz to 7500 Hz, gain 1×, and stored offline for analysis. Recordings began 5 min before and ended 5 min after the task. MedPC timestamps were integrated through the Omniplex acquisition, and animal activity was observed and recorded during the task with a video camera during the session.
5
Primate Gaze-Controlled Choice Behavior
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Monkeys were trained to sit head restrained in a custom primate chair situated 56 cm from a 19-inch monitor screen. Choices were reported using gaze location, which was monitored and acquired at 90 frames per second using an infrared oculometer (PC-60, Arrington Research, Scottsdale, AZ). Juice rewards were delivered to the monkey’s mouth using custom-made air-pressured juice dispenser systems (Mitz, 2005 (link)). Trial events, reward delivery and timings were controlled using MonkeyLogic behavioral control system (https://monkeylogic.nimh.nih.gov ), running in MATLAB (version 2014b, The MathWorks Inc.). Raw electrophysiological activity was recorded using an Omniplex data acquisition system (Plexon, Dallas, TX) and sampled at a 40kHz resolution. Spikes from putative single neurons were automatically clustered offline using the MountainSort plugin of MountainLab (Chung et al., 2017 (link)) and later curated manually based on the principal component analysis, interspike interval distributions, visually differentiated waveforms, and objective cluster measures (Isolation > 0.75, Noise overlap < 0.2, Peak signal to noise ratio > 0.5, Firing Rate > 0.05 Hz) (Stoll and Rudebeck, 2023 ).
6
Spinal Cord Neuronal Recording Protocol
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A four-electrode array with a total of 16 recording sites (A4 type, NeuroNexus) was implanted at the L5 spinal level. The array was oriented so that each electrode was at the same mediolateral position. Electrode tips were lowered 328 ± 95 μm (mean ± SD ) below the dorsal surface. Since recording sites are spaced at 50 μm intervals up each electrode, we determined on which site a unit was recorded and from this measured the depth of recorded neurons to be <300 μm (see Figure 3C ), which places them within lamina I or II (Watson et al., 2008 (link)). Neurons that responded to limb displacement, indicating proprioceptive input, were excluded. The signal was amplified, filtered at 500 Hz – 10 kHz, digitized at 20 kHz with an Omniplex Data Acquisition System (Plexon) and stored with stimulus markers on disk. Single units were isolated using Offline Sorter V3 software (Plexon), and were analyzed with Neuroexplorer 4 (Plexon). Spike waveforms were identified as monophasic or biphasic based on the absence or presence, respectively, of a negative phase (upward deflection) following the initial positive phase (see Figure 2A ).
7
Spinal Cord Neuron Recording Protocol
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A 4-shank probe with a total of 16 recording microelectrodes (A4 × 4–3mm-50-125-177-A16, NeuroNexus, Ann Arbor, MI, USA) was implanted at the L5 spinal level in the left DH. The array was oriented parallel to the rostrocaudal axis so that each electrode was at roughly the same mediolateral position. The depth of electrode insertion was monitored to estimate electrode position within the presumed Rexed lamina(e).
Electrode shank tips were lowered below the dorsal surface. Since the recording sites were spaced at 50 µm intervals up each shank of the electrode, we estimated the depth of the recorded neurons to be positioned within lamina II–III. Neurons that responded to limb displacement, indicating that they received proprioceptive input, were excluded. Measured extracellular signals of neuronal activity were amplified, and the band-pass was filtered between 500 Hz and 10 kHz, digitized at 40 kHz with the OmniPlex Data Acquisition System (Plexon, Dallas, TX, USA), and stored with stimulus markers on a disk. Single units were isolated using Offline Sorter v3 software (Plexon, Dallas, TX, USA) and analyzed with NeuroExplorer 4 (Plexon, Dallas, TX, USA).
Electrode shank tips were lowered below the dorsal surface. Since the recording sites were spaced at 50 µm intervals up each shank of the electrode, we estimated the depth of the recorded neurons to be positioned within lamina II–III. Neurons that responded to limb displacement, indicating that they received proprioceptive input, were excluded. Measured extracellular signals of neuronal activity were amplified, and the band-pass was filtered between 500 Hz and 10 kHz, digitized at 40 kHz with the OmniPlex Data Acquisition System (Plexon, Dallas, TX, USA), and stored with stimulus markers on a disk. Single units were isolated using Offline Sorter v3 software (Plexon, Dallas, TX, USA) and analyzed with NeuroExplorer 4 (Plexon, Dallas, TX, USA).
8
Optogenetic Stimulation Heart Rate
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During DMN optogenetic stimulation experiments, heart rate was monitored for the duration of stimulation. Data were acquired using the OmniPlex Data Acquisition System (Plexon Inc.) and analyzed post-hoc using Spike2 analysis software (Cambridge Electronic Design Ltd). Heart rate was recorded and expressed as beats per minute (BPM).
9
Spinal Cord Neuronal Recordings
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A 4-shank probe with a total of 16 recording microelectrodes (A4x4-3mm-50-125-177-A16, NeuroNexus) was implanted at the L5 spinal level in the left DH. The array was oriented parallel to the rostrocaudal axis so that each electrode was at roughly the same mediolateral position. Depth of electrode insertion was monitored to estimate electrode position within presumed Rexed lamina(e). Electrode shank tips were lowered 354 ± 87 μm (mean ± standard deviation) below the dorsal surface. Since recording sites are spaced at 50 μm intervals up each shank of the electrode, we estimated the depth of recorded neurons to be positioned within lamina I or II. Neurons that responded to limb displacement, indicating that they received proprioceptive input, were excluded. Measured extracellular signals of neuronal activity were amplified, band-pass filtered between 500 Hz and 10 kHz, digitized at 40 kHz with OmniPlex Data Acquisition System (Plexon, Dallas, TX, USA), and stored with stimulus markers on disk. Single units were isolated using Offline Sorter v3 software (Plexon) and were analyzed with NeuroExplorer 4 (Plexon).
10
Spike Sorting of Neuronal Signals
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