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Neuroscience

Low-Cost Electroencephalographic Recording System Combined with a Millimeter-Sized Coil to Transcranially Stimulate the Mouse Brain In Vivo

Published: May 26th, 2023

DOI:

10.3791/65302

1Bioengineering and Bioinformatics, Graduate School of Information Science and Technology, Hokkaido University, 2Bioengineering and Bioinformatics, Division of Information Science and Technology, Hokkaido University
* These authors contributed equally

A low-cost electroencephalographic recording system combined with a millimeter-sized coil is proposed to drive transcranial magnetic stimulation of the mouse brain in vivo. Using conventional screw electrodes with a custom-made, flexible, multielectrode array substrate, multi-site recording can be carried out from the mouse brain in response to transcranial magnetic stimulation.

A low-cost electroencephalographic (EEG) recording system is proposed here to drive transcranial magnetic stimulation (TMS) of the mouse brain in vivo, utilizing a millimeter-sized coil. Using conventional screw electrodes combined with a custom-made, flexible, multielectrode array substrate, multi-site recording can be carried out from the mouse brain. In addition, we explain how a millimeter-sized coil is produced using low-cost equipment usually found in laboratories. Practical procedures for fabricating the flexible multielectrode array substrate and the surgical implantation technique for screw electrodes are also presented, which are necessary to produce low-noise EEG signals. Although the methodology is useful for recording from the brain of any small animal, the present report focuses on electrode implementation in an anesthetized mouse skull. Furthermore, this method can be easily extended to an awake small animal that is connected with tethered cables via a common adapter and fixed with a TMS device to the head during recording.The present version of the EEG-TMS system, which can include a maximum of 32 EEG channels (a device with 16 channels is presented as an example with fewer channels) and one TMS channel device, is described. Additionally, typical results obtained by the application of the EEG-TMS system to anesthetized mice are briefly reported.

Transcranial magnetic stimulation (TMS) is a promising tool for human brain science, clinical application, and animal model research because of its non-/low invasiveness. During the early stage of TMS applications, measurement of the cortical effect in response to single- and paired-pulse TMS in humans and animals was restricted to the motor cortex; easily measurable output was limited to motor evoked potentials and induced myoelectric potentials involving the motor cortex1,2. To expand the brain regions that can be measured by TMS modulation, electroencephalographic (EEG) recording was integrated with single-....

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In the present study, all animal experiments were performed following the National Institutes of Health Guide for the Care and Use of Laboratory Animals and with approval from the Institutional Animal Care and Use Committee of Hokkaido University. C57BL/6J mice, two male and three female, 8 to 10 weeks old, were used for the present study. This is a terminal procedure. The animals were obtained from a commercial source (see Table of Materials).

1. Flexible two-dimensiona.......

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Sample EEG data recorded in anesthetized C57BL/6J mice with the flexible substrate combined with the screw electrodes are presented below.

As a typical example, the average EEG waveforms generated in response to sound stimulation (8 kHz tone-burst, 80 dB sound pressure level [SPL]) are shown for 60 trials with identical stimuli (Figure 4A). A schematic of recording channel mapping is also presented in the middle of Figure 4A. The resp.......

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This study addresses a multi-site EEG recording system combined with a magnetic stimulation system designed for small animals, including mice. The constructed system is low-cost and easily constructed in physiological laboratories, and can extend their existing measurement setups. The surgical procedure necessary to obtain data from the mouse recording system is profoundly simple if such laboratories have previous experience with standard electrophysiological experiments.

One advantage of usin.......

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This work was supported by the Murata Science Foundation, the Suzuken Memorial Foundation, the Nakatani Foundation for Advancement of Measuring Technologies in Biomedical Engineering, and a Grant-in-Aid for Exploratory Research (grant number 21K19755, Japan) and for Scientific Research (B) (grant number 23H03416, Japan) to T.T.

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Name Company Catalog Number Comments
3D printer Zhejiang Flashforge 3D Technology Co., Ltd FFD-101 The printer used for 3D-printing the donut-shaped disks
ATROPINE SULFATE  0.5 mg NIPRO ES PHARMA CO., LTD. - Atropine sulfate
Bipolar amplifier NF Corp. KIT61380 For amplifying waveforms for coil input
Butorphanol Meiji Seika Pharma
Co., Ltd., Tokyo, Japan
- For anathesis of animals
Commercial manufacturer of flexible 2D array p-ban.com Corp. - URL: https://www.p-ban.com/
Computer prograom to analyze output signals Natinal Instruments NI-DAQ and  NI-DAQmx Python To analyze output signals from the hall-effect sensor
Connector Harwin Inc. G125-FV12005L0P For connector to conect to the measuring system
Copper pad p-ban.com Corp. copper Copper pad on each substrate
Copper wire Kyowa Harmonet Ltd. P644432 The windings of the coil
DAQ board National Instruments Corp. USB-6343 For measuring the magnitic flux density of the coil
Dental cement SHOFU INC. Quick Resin Self-Curing Orthodontic Resin
ECoG electrode NeuroNexus Inc. HC32 For reference to design of the flexible 2D array
Epoxy resin Konishi Co. Ltd. #16123 For coil construction
Ethyl Carbamate FUJIFILM Wako Pure Chemical Corp. 050-05821 For urethan anesthesia
Flat ribbon cable Oki Electric Cable Co., Ltd. FLEX-B2(20)-7/0.1 20028 5m For cable to connect between surface-mount connector and measuring sysytem
flexible substrate p-ban.com Corp. polyimide Baseplate of flexible substrate
Function generator NF Corp. WF1947 For generating waveforms for coil input
Hall-effect sensor Honeywell International Inc. SS94A2D For measuring the magnitic flux density of the coil
IDC crimping tool Pro'sKit Industries Co. 6PK-214 To crimp the IDC and one end of the flat ribbon cable; Flat cable connector crimping tool
Instant glue Konishi Co. Ltd. #04612 For coil construction
Insulation-displacement connector (IDC ) Uxcell Japan B07GDDG3XG 2 × 10 pins and a 1.27 mm pitch 
LCR meter NF Corp. ZM2376 For measuring the AC properties of the coil
Manipulator NARISHIGE Group. SM-15L For manipulating the coil
Medetomidine Kobayashi Kako, Fukui, Japan - For anathesis of animals
Midazolam Astellas Pharma, Tokyo, Japan - For anathesis of animals
Miniature screw KOFUSEIBYO Co., Ltd. S0.6*1.5 For EEG-senseing and reference electrode
Mouse Japan SLC, Inc. C57BL/6J (C57BL/6JJmsSlc) Experimental animal
Permalloy-45 rod The Nilaco Corp. 780544 The core of the coil
Recording system Plexon Inc. OmniPlex For EEG data acquisition
Stainless wire Wakisangyo Co., Ltd. HW-136 For grasp by manipulator
Stereotaxic apparatus NARISHIGE Group. SR-5M-HT To fix a mouse head
Surface-mount connector Useconn Electronics Ltd. PH127-2x10MG For connector to mount on the flexible 2D array
Testing equipment (LCR meter) NF Corp. ZM2372 Contact check and impedance measurements
White PLA filament Zhejiang Flashforge 3D Technology Co., Ltd PLA-F13 The material used for 3D-printing the donut-shaped disks
Xylocaine Jelly 2% Sandoz Pharma Co., Ltd. - lidocaine hydrochloride

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