a micro-drive array method for electrophysiological recording from multiple brain regions

A Micro-drive Array Method for Electrophysiological Recording from Multiple Brain Regions

Secure the rat to a stereotaxic device, then, shave and clean the skull, and start the array implantation with a 3-centimeter incision along the midline from the point between the eyes to the neck. Using a high-speed drill, make a pair of circular craniotomies between 0.7 and 1.0 millimeters in diameter above the olfactory bulb, which is 11.0 millimeters anterior and one millimeter bilateral to bregma.
Then, implant the two BR electrodes just deep enough for the tips of the screws to make contact with the brain, which is about 2 millimeters deep. Use between six and eight full turns of the screw.
Next, make another pair of craniotomies above the frontal cortex, 2.7 millimeters anterior and 2.7 millimeters bilateral to bregma. Into these holes, secure the two ground reference electrodes so they also just contact the brain, about 1.6 millimeters deep. This takes four to five full turns of the screw.
Now, plan a large circular craniotomy with a diameter of about 2.0 millimeters above the hippocampus, 3.8 millimeters posterior, and 2.5 millimeters bilateral to bregma. Then, make six to eight 1.0-millimeter holes in the area that will surround the craniotomy.
Into each of these holes, implant the anchor screws. Then, make the planned craniotomy between the screws. Above the large hole, position the integrative microdrive array such that the cannula tip is just above the large craniotomy.
Fill the gap space between the cannula tip and the brain surface with about 100 microliters of two-part epoxy. Over the next five minutes, allow the mixture to change into a transparent gel. Next, cover the cannula, BR electrodes, ground reference electrodes, and anchor screws with dental cement.
Apply about five millimeters of cement without covering the open ends of BR or ground reference electrodes. Now, solder all the open ends of the electrodes to where they connect to the board. Then, cover the bottom part of the integrative microdrive array and all electrode wires with dental cement.
It is important to completely cover the electrode wires so that the rat cannot scratch them out after the implantation. Now allow the rat to recover. After regaining sufficient consciousness to maintain sternal recumbency, house it solo without any cage mates, and give it free access to food and water.
After the surgery, gradually advance the tetrodes by advancing the screws daily. Once the tetrodes are adjacent to the target brain areas, let them settle for several days during which signals will stabilize.

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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Preparation of the Integrative Micro-Drive Array
<ol>
	<li>Create a micro-drive array for cortical local field potential (LFP) recordings as described elsewhere. Leave at least 6 metal holes open on an electrode interface board (EIB) for use as electrocardiography/electromyography/breathing rate (ECG/EMG/BR) channels that are connected to bioflex wires as described in 1.2.</li>
	<li>Cut a bioflex wire into 6 pieces with lengths of 5.0 cm. Peel off the polytetrafluoroethylene (PTFE) coating of both ends of all the wire pieces with lengths of ~5.0 mm. Connect one end of each of the wire pieces to one of the open metallic holes (ECG/EMG/BR channels) on the EIB with a gold pin.</li>
	<li>Cut an enamel wire into two 5.0-cm pieces. Solder one end of each of these wires to the ground/reference (g/r) channels on the EIB (Figure 1).</li>
	<li>For the preparation of ECG electrodes, cut a bioflex wire into two 16-cm pieces. Peel off the PTFE coating of the ends of these wire pieces at lengths of ~5.0 mm on one end (short end) and ~15 mm on the other end (long end).</li>
	<li>Form a wire ring with a diameter of 2.0 mm by bending the long end of the wire and fixing the shape of the ring by soldering.</li>
	<li>For the preparation of EMG electrodes, cut a bioflex wire into 2 pieces with lengths of 8 cm. Peel off the PTFE coating from both ends of these wire pieces with lengths of ~5.0 mm.</li>
	<li>For the preparation of BR electrodes, cut a bioflex wire into 2 pieces with lengths of 6.0 cm. Peel off the enamel coating of both ends of these wire pieces with lengths of ~5.0 mm. Solder one end of each of these wire pieces to the head of a stainless- steel screw (stem diameter: 1.0 mm, stem length: 4.0 mm).</li>
	<li>For the preparation of ground/reference (gr) electrodes, cut an enamel wire into 2 pieces with lengths of 6.0 cm. Peel off the enamel coating of both ends of these wire pieces with lengths of ~5.0 mm. Solder one end of each of these wire pieces to the head of a stainless- steel screw (stem diameter: 1.4 mm, stem length: 3.0 mm).</li>
	<li>Gas sterilizes all electrodes and stainless screws and keeps these in a clean space.</li>
</ol>
2. Implantation of the Integrative Micro-Drive Array and the BR Electrodes
<ol>
	<li>Fix the rat on a stereotaxic device. Make an incision of ~3.0 cm on the head along the midline from the point between the eyes to the neck area. Expose the skull.</li>
	<li>Make two circular craniotomies with diameters of 0.7-1.0 mm above the olfactory bulb 11.0 mm anterior and 1 mm bilateral to bregma with a high-speed drill. Implant two BR electrodes in the skull until the tips of the screw stems are attached to the brain surface.</li>
	<li>Make two circular craniotomies with diameters of 0.7-1.0 mm above the frontal cortex 2.7 mm anterior and 2.7 mm bilateral to bregma. Implant two g/r electrodes in the skull until the tip of the screw stem is attached to the brain surface.</li>
	<li>Make six to eight holes with diameters of 1.0 mm in the area surrounding the large craniotomy. Implant anchor screws (stem diameter: 1.4 mm, stem length: 3.0 mm) in the skull.</li>
	<li>Make a large circular craniotomy with a diameter of ~2.0 mm above the hippocampus 3.8 mm posterior and 2.5 mm bilateral to bregma. Place the integrative micro-drive array such that the cannula tip of the drive array is located above the large craniotomy</li>
	<li>Fill the gap space between the cannula tip and the brain surface with ~100 &#181;L of two solutions, i.e., 0.5% (by mass) sodium alginate and 10% (by mass) calcium chloride. 
	NOTE: This process forms a transparent gel in ~5 min, after the two solutions are mixed on the skull.</li>
	<li>Cover the cannula, BR electrodes, g/r electrodes, and anchor screws with dental cement with a thickness of 0.5 cm. Be careful NOT to cover the open ends of BR and g/r electrodes with the cement at this step.</li>
	<li>Solder the open ends of the ECG, EMG, BR and g/r electrodes to the individual wire tips that were previously connected to the EIB (see the steps 1.2 and 1.3).</li>
	<li>Cover the bottom part of the integrative micro-drive array, and all electrode wires, with dental cement. Ensure that all the electrode wires are completely covered so that the rat cannot scratch them out after the implantation.</li>
	<li>After regaining sufficient consciousness to maintain sternal recumbency, return the animal to its transparent Plexiglas home cage, and keep it on its own with free access to water and food. After the surgery, treat the animal with antibiotics (gentamicin).</li>
</ol>
3. In Vivo Recordings
NOTE: All signals are amplified, sampled at 2 kHz, and band-pass filtered (0.1 - 500 Hz) except for unit activities (sampled at 30 kHz and bandpass filtered (500 - 6 kHz)).
<ol>
	<li>Connect the EIB of the integrative micro-drive array to the headstage of a recording device.</li>
	<li>Advance the tetrodes by turning the screws for a few weeks after surgery. Once the tetrodes are adjacent to the target brain areas, settle the tetrodes into the areas over a period of several days for stable recordings.</li>
	<li>Monitor the electrical signals while the animal freely moves in a recording chamber.</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>FEP Hookup Wire Stranded Stainless Steel</td>
			<td>Cooner Wire Company, Chatsworth, CA</td>
			<td>AS 633</td>
			<td>Bioflex wire</td>
		</tr>
		<tr>
			<td>EIB-36-PTB</td>
			<td>Neuralynx, Inc., Bozeman, MT</td>
			<td>EIB-36-PTB</td>
			<td>EIB</td>
		</tr>
		<tr>
			<td>Cereplex M</td>
			<td>Blackrock Microsystems, Salt Lake City, UT</td>
			<td></td>
			<td>Digital headstage</td>
		</tr>
		<tr>
			<td>Cereplex Direct</td>
			<td>Blackrock Microsystems, Salt Lake City, UT</td>
			<td></td>
			<td>Data acquisition system</td>
		</tr>
		<tr>
			<td>UEW polyurethane magnet wire</td>
			<td>Oyaide.com, Tokyo, Japan</td>
			<td>UEW 0.14mm 20m</td>
			<td>Enamel wire</td>
		</tr>
		<tr>
			<td>SD-102</td>
			<td>Narishige, Tokyo, Japan</td>
			<td>SD-102</td>
			<td>High-speed drill</td>
		</tr>
		<tr>
			<td>Minimo ONE SERIES ver.2</td>
			<td>Minitor Co.,Ltd, Tokyo, Japan</td>
			<td>C2012</td>
			<td>High-peed drill Power Supply</td>
		</tr>
		<tr>
			<td>Provinice 250 mL</td>
			<td>Shofu Inc., Kyoto, Japan</td>
			<td>213620136</td>
			<td>Dental cement</td>
		</tr>
		<tr>
			<td>Small Animal Anesthetizer</td>
			<td>Biomachinery, Chiba, Japan</td>
			<td>TK-7</td>
			<td>Anesthetizer</td>
		</tr>
		<tr>
			<td>Buprenorphine hydrochloride</td>
			<td>Sigma-Aldrich, St. Louis, MO</td>
			<td>B7536-1ML</td>
			<td>Analgesic</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>DS Pharma Animal Health, Osaka, Japan</td>
			<td>Isoflu 250mL</td>
			<td></td>
		</tr>
		<tr>
			<td>Vaseline, White</td>
			<td>Wako Pure Chemical Industries, Ltd., Osaka, Japan</td>
			<td>224-00165</td>
			<td>Vet ointment</td>
		</tr>
		<tr>
			<td>Sodium alginate</td>
			<td>Nacalai tesque, Kyoto, Japan</td>
			<td>31131-85</td>
			<td></td>
		</tr>
		<tr>
			<td>Calcium Chloride Dihydrate</td>
			<td>Wako Pure Chemical Industries, Ltd., Osaka, Japan</td>
			<td>031-00435</td>
			<td></td>
		</tr>
		<tr>
			<td>Stainless steel screw M1.0&#215;4.0</td>
			<td>MonotaRO, Hyogo, Japan</td>
			<td>42617504</td>
			<td>Stainless steel screw for BR electrodes</td>
		</tr>
		<tr>
			<td>Stainless steel screw M1.4&#215;3.0</td>
			<td>MonotaRO, Hyogo, Japan</td>
			<td>42617687</td>
			<td>Stainless steel screw for g/r electrodes and anchors</td>
		</tr>
	</tbody>
</table>

Source: Shikano, Y., et al. <a href="https://app.jove.com/v/56980">Simultaneous Recordings of Cortical Local Field Potentials, Electrocardiogram, Electromyogram, and Breathing Rhythm from a Freely Moving Rat.</a> J. Vis. Exp. (2018).
The video demonstrates simultaneous electrophysiological recording from multiple brain regions in a rat using an integrative micro-drive array. Recording electrodes are placed in the olfactory bulb, reference electrodes in the frontal cortex, and micro-drive array electrodes in the hippocampus. The rat is allowed to forage freely, and simultaneous recording from all the implanted electrodes is recorded using the array.

<img alt="Figure 1" src="/files/ftp_upload/22834/22834_Figure1.jpg" /> 
Figure 1: A top view of the EIB. The EIB includes 24 cortical LFP (LFP) channels that are connected to the tetrodes, 2 ECG channels, 2 EMG channels, 2 BR channels, and 2 ground (Gr) channels. All channels except the LFP channels were connected to insulated wires.

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

Secure an anesthetized rat in a stereotaxic frame. Shave and disinfect the scalp, then make an incision to expose the skull.
Drill holes above the olfactory bulb to implant recording electrodes. Drill above the frontal cortex to implant reference electrodes that acquire baseline brain activity signals.
Insert anchor screws around the hippocampus and create a large hole above the region.
Position an integrative micro-drive array with its cannula containing recording electrodes above the hole.
Use a gel-forming solution to secure the array. Apply dental cement that adheres to the anchor screws and provides mechanical stability.
Connect the array to the implanted electrodes for simultaneous recording from multiple brain regions. Insulate the connections to enhance signal quality.
Allow the rat to recover, then advance the array electrodes to contact the brain and let the signals stabilize.
The rat with the implanted electrodes is ready to record electrical signals from the brain.

18 vues. Procédé de micro-réseau d’entraînement pour l’enregistrement électrophysiologique de plusieurs régions du cerveau

Vidéo: Procédé de micro-réseau d’entraînement pour l’enregistrement électrophysiologique de plusieurs régions du cerveau - Expérience

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such as epilepsy and neurodegenerative disorders. However, it is technically difficult to obtain EEG recordings in neonates as it requires specialized handling and great care. Here, we present a novel method to record EEG in neonatal rat pups (P8-P15). We designed a simple and reliable electrode using computer pin loci; it can be easily implanted into the skull of a rat pup to record high-quality EEG signals in the normal and epileptic brain. Pups were given an intraperitoneal (i.p.) injection of the neurotoxin kainic acid (KA) to induce epileptic seizures. The surgical implantation performed in this procedure is less expensive than other EEG procedures for neonates. This method allows one to record high-quality and stable EEG signals for more than 1 week. Furthermore, this procedure can also be applied to adult rats and mice to study epilepsy or other neurological disorders.

Here, we introduce a novel technique designed to record electroencephalography (EEG) in freely moving neonatal epileptic pups and describe its procedures, features, and applications. This method allows one to record EEG for more than 1 week.

Enregistrement d&#39;EEG dans des rats néonatals à déplacement libre à l&#39;aide d&#39;une nouvelle méthode

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such ...

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Evaluation of Hemisphere Lateralization with Bilateral Local Field Potential Recording in Secondary Motor Cortex of Mice

This article demonstrates complete, detailed procedures for both in vivo bilateral recording and analysis of local field potential (LFP) in the cortical areas of mice, which are useful for evaluating possible laterality deficits, as well as for assessing brain connectivity and coupling of neural network activities in rodents. The pathological mechanisms underlying Alzheimer&#39;s disease (AD), a common neurodegenerative disease, remain largely unknown. Altered brain laterality has been demonstrated in aging people, but whether or not abnormal lateralization is one of the early signs of AD has not been determined. To investigate this, we recorded bilateral LFPs in 3-5-month-old AD model mice, APP/PS1, together with littermate wild type (WT) controls. The LFPs of the left and right secondary motor cortex (M2), specifically in the gamma band, were more synchronized in APP/PS1 mice than in WT controls, suggesting a declined hemispheric asymmetry of bilateral M2 in this AD mouse model. Notably, the recording and data analysis processes are flexible and easy to carry out, and can also be applied to other brain pathways when conducting experiments that focus on neuronal circuits.

We present in vivo electrophysiological recording of the local field potential (LFP) in bilateral secondary motor cortex (M2) of mice, which can be applied to evaluate hemisphere lateralization. The study revealed altered levels of synchronization between the left and right M2 in APP/PS1 mice compared to WT controls.

Évaluation de la latéralisation de l'hémisphère avec enregistrement potentiel de terrain local bilatéral dans le cortex moteur secondaire des souris

This article demonstrates complete, detailed procedures for both in vivo bilateral recording and analysis of local field potential (LFP) in the cortical ...

Construction of Local Field Potential Microelectrodes for in vivo Recordings from Multiple Brain Structures Simultaneously

Researchers often need to record local field potentials (LFPs) simultaneously from several brain structures. Recording from multiple desired brain regions requires different microelectrode designs, but commercially available microelectrode arrays often do not offer such flexibility. Here, the present protocol outlines the straightforward design of custom-made microelectrode arrays to record LFPs from multiple brain structures simultaneously at different depths. This work describes the construction of the bilateral cortical, striatal, ventrolateral thalamic, and nigral microelectrodes as an example. The outlined design principle offers flexibility, and the microelectrodes can be modified and customized to record LFPs from any structure by calculating stereotaxic coordinates and quickly changing the construction accordingly to target different brain regions in either freely moving or anesthetized mice. The microelectrode assembly requires standard tools and supplies. These custom microelectrode arrays allow investigators to easily design microelectrode arrays in any configuration to track neuronal activity, providing LFP recordings with millisecond resolution.

Construction of Local Field Potential Microelectrodes for in vivo Recordings from Multiple Brain Structures Simultaneously

The present protocol describes the construction of custom-made microelectrode arrays to record local field potentials in vivo from multiple brain structures simultaneously.

Construction de microélectrodes de potentiel de champ local pour des enregistrements in vivo à partir de plusieurs structures cérébrales simultanément

Researchers often need to record local field potentials (LFPs) simultaneously from several brain structures. Recording from multiple desired brain regions ...

A Micro-drive Array Method for Electrophysiological Recording from Multiple Brain Regions

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