LFP recordings were taken in parallel with the imaging sessions. LFPs were recorded using an RHD2000 multi-channel amplifier (Intan Technologies) acquired with RHD2000 interface software (version 1.5, copyright Intan Technologies). LFP recordings were acquired at 20 kHz, low-pass filtered (eighth order Chebyshev Type I IIR filter with cutoff frequency at 80 Hz and zero-phase distortion), and downsampled to 200 Hz.
Rhd2000 interface software
Manufactured by Intan Technologies
Sourced in United States
The RHD2000 interface software is a digital data acquisition system designed to interface with various types of lab equipment. It provides a user-friendly platform for capturing and analyzing data from compatible devices. The software offers real-time data processing and visualization capabilities, enabling users to monitor and record experimental parameters.
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4 protocols using rhd2000 interface software
1
Multimodal Recording of Local Field Potentials
2
Electrophysiology and Optogenetic Stimulation
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The arrays were connected to an Intan RHD2216 amplifier board, which in turn was connected to a RHD2000 SPI interface cable (Intan, Los Angeles, CA). The cable connected to a custom-designed Hall effect-based active commutator (NeuroTek Innovative Technology Incorporated, Toronto, Ontario, Canada). The commutator fed this digitized data into the RHD2000 USB board (Intan). Data were recorded at 25 kHz continuously for up to 100 hr using the RHD2000 Interface Software (Intan). Spike extraction, clustering, and sorting were done using custom MATLAB and Python code (see below). Light for stimulation was produced using a fiber coupled 470 nm LED system (M470F3/LEDD1B, Thorlabs). Light irradiance was adjusted for each animal and never exceeded 18 mW/cm2. Stimulation was triggered using voltage pulses of varying magnitude from a NIDAQ board (controlled by custom MATLAB program) to the LED driver, set to modulation mode.
3
Hippocampal LFP Dynamics and Calcium Signaling
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In parallel to calcium fluorescence events, we recorded the local field potentials (LFP) from the CA3 region of the hippocampus using a microarray with eight bipolar electrodes (MicroProbes). Data were recorded through a RHD2000 multichannel amplifier with RHD2000 interface software (v1.5, Intan Technologies) at a 20 kHz sample rate and imported in MATLAB (R2020b, MathWorks, Inc.). The raw data were lowpass filtered (20th order FIR filter, cutoff frequency 1.3 kHz), and the resultant signals were further filtered (cutoff frequency: 80 Hz, 8th order Chebyshev Type I lowpass filter applied in both directions to avoid phase distortions) before being down-sampled to 200 Hz. For each recording, 30 s-long segments were selected for spectral analyses. LFP channels in each segment were standardized and processed via continuous wavelet transform (10 voices per octave, analytic bump wavelet) to perform a time-frequency analysis. All analyses were performed blind to the genotype.
4
Electrochemical impedance of carbon fiber electrodes
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The electrochemical impedances of carbon fiber electrodes before and after fire-sharpening process was measured at 1 kHz in phosphate-buffered saline (PBS) solution using a 3-electrode based potentiostat system (ZIVE SP2; ZIVE lab., Seoul, Korea) with Ag/AgCl electrode (MF-2052; Bioanalytical Systems, Inc., West Lafayette, IN, USA) as a reference electrode and Pt electrode (CHI115; CH Instruments, Inc., Austin, TX, USA) as a counter electrode. After electrode implantation into the rat brain, the impedances of the carbon fiber electrode embedded with silk and tungsten supports were measured every two days for 1 month. Since there was no counter electrode, the impedance was measured using 2-electrode configuration with the implanted silver wire as a reference electrode and the carbon fiber as a working electrode using RHD2000 interface software (Intan technologies, Los Angeles, CA, USA). The impedance magnitudes measured at 1 kHz were compared between tungsten- and silk-support electrode arrays.
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