Name: construction of local field potential microelectrodes for in vivo recordings from multiple brain structures simultaneously
Uploaded: 2022-03-14T15:00:00
Description: Watch this Scientific Journal Video about Construction of Local Field Potential Microelectrodes for in vivo Recordings from Multiple Brain Structures Simultaneously at JoVE.com

这项工作得到了美国国立卫生研究院（RO1 NS120945，R37NS119012至JK）和UVA脑研究所的支持。

本工作已获得弗吉尼亚大学动物护理和使用委员会的批准。C57Bl / 6两性小鼠（7-12周）用于实验。这些动物保持在12小时光照/ 12小时黑暗循环中，并 可以随意 获得食物和水。
1. 微电极结构
<ol><li>要构建微电极，请使用50μm（直径）的透镜涂层镍铬丝（见 材料表）。在平台背面用胶带将电线的一端缠绕在平台上最近的旋钮上三次（<strong class="xf...

<ol>
	<li>Buzs&#225;ki, G., Anastassiou, C. A., Koch, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+origin+of+extracellular+fields+and+currents-EEG,+ECoG,+LFP+and+spikes.">The origin of extracellular fields and currents-EEG, ECoG, LFP and spikes.</a> Nature Reviews Neuroscience. 13, 407-420 (2012).</li><li>Hubel, D. H., Wiesel, T. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Receptive+fields+of+single+neurones+in+the+cat's+striate+cortex.">Receptive fields of single neurones in the cat's striate cortex.</a> The Journal of Physiology. 148 (3), 574-591 (1959).</li><li>O'Keefe, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Place+units+in+the+hippocampus+of+the+freely+moving+rat.">Place units in the hippocampus of the freely moving rat.</a> Experimental Neurology. 51 (1), 78-109 (1976).</li><li>Fyhn, M., Molden, S., Witter, M. P., Moser, E. I., Moser, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+representation+in+the+entorhinal+cortex.">Spatial representation in the entorhinal cortex.</a> Science. 305 (5688), 1258-1264 (2004).</li><li>Buzs&#225;ki, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Large-scale+recording+of+neuronal+ensembles.">Large-scale recording of neuronal ensembles.</a> Nature Neuroscience. 7, 446-451 (2004).</li><li>Buckmaster, P. S., Edward Dudek, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+intracellular+analysis+of+granule+cell+axon+reorganization+in+epileptic+rats.">In vivo intracellular analysis of granule cell axon reorganization in epileptic rats.</a> Journal of Neurophysiology. 81 (2), 712-721 (1999).</li><li>Driscoll, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multimodal+in+vivo+recording+using+transparent+graphene+microelectrodes+illuminates+spatiotemporal+seizure+dynamics+at+the+microscale.">Multimodal in vivo recording using transparent graphene microelectrodes illuminates spatiotemporal seizure dynamics at the microscale.</a> Communications Biology. 4, 1-14 (2021).</li><li>Roy, D. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Memory+retrieval+by+activating+engram+cells+in+mouse+models+of+early+Alzheimer's+disease.">Memory retrieval by activating engram cells in mouse models of early Alzheimer's disease.</a> Nature. 531, 508-512 (2016).</li><li>Igarashi, K. M., Lu, L., Colgin, L. L., Moser, M. B., Moser, E. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coordination+of+entorhinal-hippocampal+ensemble+activity+during+associative+learning.">Coordination of entorhinal-hippocampal ensemble activity during associative learning.</a> Nature. 510, 143-147 (2014).</li><li>Gage, G. J., Kipke, D. R., Shain, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole+animal+perfusion+fixation+for+rodents.">Whole animal perfusion fixation for rodents.</a> Journal of Visualized Experiments. (65), e3564 (2012).</li><li>Brodovskaya, A., Shiono, S., Kapur, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activation+of+the+basal+ganglia+and+indirect+pathway+neurons+during+frontal+lobe+seizures.">Activation of the basal ganglia and indirect pathway neurons during frontal lobe seizures.</a> Brain. 144 (7), 2074-2091 (2021).</li><li>Ren, X., Brodovskaya, A., Hudson, J. L., Kapur, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Connectivity+and+neuronal+synchrony+during+seizures.">Connectivity and neuronal synchrony during seizures.</a> The Journal of Neuroscience. 41 (36), 7623-7635 (2021).</li><li>Chang, E. H., Frattini, S. A., Robbiati, S., Huerta, P. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Construction+of+microdrive+arrays+for+chronic+neural+recordings+in+awake+behaving+mice.">Construction of microdrive arrays for chronic neural recordings in awake behaving mice.</a> Journal of Visualized Experiments. (77), e50470 (2013).</li></ol>

从历史上看，微电极阵列已被广泛用于记录来自感兴趣的特定大脑区域2，3，4，5，6，7，8，9，13的神经元活动。但是，我们简单的微电极设计允许同时从多个结构记录<s...

局部场电位（LFP）是从大脑细胞外空间记录的电势。它们由神经元外部的离子浓度失衡产生，代表一小群局部神经元的活动，与宏观尺度的脑电图记录相比，可以精确地监测特定大脑区域的活动1。据估计，相隔1毫米的LFP微电极对应于两个完全不同的神经元群体。脑电图信号由脑组织、脑脊液、颅骨、肌肉和皮肤过滤，而LFP信号是局部神经元活动的可靠标志物1。
研究人员通常需要同时记录来自几种大脑结构的LFP，但市售的微电极阵列通常不提供这种灵活性。在这里，本方案描述了完全可定制，易于构建的微电极，以同时记录来自不同深度的任何所需大脑区域的LFP。虽然LFPs已被广泛用于记录特定大脑区域2，3，4，5，6，7，8，9的神经元活动，但当前易于定制的设计允许记录来自任何多个浅表或深部大脑区域的LFP11，12.还可以修改该协议，通过确定大脑区域的立体定位坐标并相应地组装阵列来构建任何所需的微电极阵列。这些微电极具有10 kHz采样率和60-70 kΩ电阻（2 cm长度），使我们能够以毫秒级精度记录LFP。然后，数据可以通过16通道放大器放大，滤波（低通1 Hz，高通5 kHz）并数字化。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Amplifier 16-Channel</td>
			<td>A-M Systems</td>
			<td>Model 3600</td>
			<td>Amplifier</td>
		</tr>
		<tr>
			<td>Cranioplasty cement</td>
			<td>Coltene</td>
			<td>Perm Reeline/Repair Resin Type II Class I Shade - Clear</td>
			<td>Cement to hold microelectrodes</td>
		</tr>
		<tr>
			<td>Cryostat Microtome</td>
			<td>Precisionary</td>
			<td>CF-6100</td>
			<td>To slice brain</td>
		</tr>
		<tr>
			<td>Diamel-coatednickel-chromium wire</td>
			<td>Johnson Matthey Inc.</td>
			<td>50 &#181;m</td>
			<td>Microelectrode wire</td>
		</tr>
		<tr>
			<td>Dremel</td>
			<td>Dremel</td>
			<td>300 Series</td>
			<td>To drill holes in mouse skull</td>
		</tr>
		<tr>
			<td>Epoxy</td>
			<td>CEC Corp</td>
			<td>C-POXY 5</td>
			<td>Fast setting adhesive</td>
		</tr>
		<tr>
			<td>Hemostat</td>
			<td>Any</td>
			<td></td>
			<td>To hold the headset</td>
		</tr>
		<tr>
			<td>Forceps</td>
			<td>Any</td>
			<td></td>
			<td>To hold microelectrodes</td>
		</tr>
		<tr>
			<td>Light microscope</td>
			<td>Nikon</td>
			<td>SMZ-10</td>
			<td>To see alignment</td>
		</tr>
		<tr>
			<td>Ohmmeter</td>
			<td>Any</td>
			<td></td>
			<td>To measurre resistance</td>
		</tr>
		<tr>
			<td>Pins (Headers and matching Sockets)</td>
			<td>Mill-Max</td>
			<td>Interconnects, 833 series, 2 mm grid gull wing surface mount headers and sockets</td>
			<td>To attach microelectrodes to</td>
		</tr>
		<tr>
			<td>Polymicro Tubing Kit</td>
			<td>Neuralynx</td>
			<td>ID 100 &#177; 04 &#181;m, OD 164 &#177; 06 &#181;m, coating thickness 12 &#181;m</td>
			<td>Glass tubes</td>
		</tr>
		<tr>
			<td>Pulse Stimulator</td>
			<td>A-M Systems</td>
			<td>Model 2100</td>
			<td>To mark the microelectrode location at the end of the recordings</td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td>Any</td>
			<td></td>
			<td>To cut microelectrodes</td>
		</tr>
		<tr>
			<td>Superglue</td>
			<td>Gorilla</td>
			<td></td>
			<td>Adhesive</td>
		</tr>
		<tr>
			<td>Thick wire 0.008 in. &#8211; 0.011 in.</td>
			<td>A-M Systems</td>
			<td>791900</td>
			<td>Tick wire to hold the microelectrode array</td>
		</tr>
		<tr>
			<td>Thin wire 0.005 in. - 0.008 in.</td>
			<td>A-M Systems</td>
			<td>791400</td>
			<td>Thin wire for reference and ground</td>
		</tr>
	</tbody>
</table>

construction of local field potential microelectrodes for in vivo recordings from multiple brain structures simultaneously

研究人员通常需要同时记录来自几种大脑结构的局部场电位（LFP）。从多个所需的大脑区域进行记录需要不同的微电极设计，但市售的微电极阵列通常不提供这种灵活性。在这里，本协议概述了定制微电极阵列的简单设计，以在不同深度同时记录来自多个大脑结构的LFP。这项工作描述了双侧皮质，纹状体，腹外侧丘脑和黑质微电极的构造作为示例。概述的设计原理提供了灵活性，微电极可以通过计算立体定位坐标并相应地快速更改结构以针对自由移动或麻醉小鼠中的不同大脑区域进行修改和定制，以记录任何结构中的LFP。微电极组件需要标准工具和耗材。这些定制的微电极阵列使研究人员能够轻松设计任何配置的微电极阵列来跟踪神经元活动，从而提供毫秒级分辨率的LFP记录。

在这项工作中，LFP微电极用于绘制通过基底神经节11扩散的癫痫发作。从右侧运动前皮层（癫痫发作焦点所在）和左侧VL，纹状体和SNR同时进行LFP记录（图4）。癫痫发作开始被确定为电压迹线的偏转至少是基线的两倍（图4A，红色箭头）。功率谱图11显示了记录的LFP的频率分布（图4A）。可以以?...

Watch this Scientific Journal Video about Construction of Local Field Potential Microelectrodes for in vivo Recordings from Multiple Brain Structures Simultaneously at JoVE.com

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

本方案描述了定制微电极阵列的构建，以同时记录来自多个大脑结构 的体内 局部场电位。

构建局部场势微电极，用于同时从多个脑结构 进行体内 记录

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

研究人员通常需要同时记录来自多个结构的局部场电位。我们简单的微电极设计使我们能够同时记录来自不同深度的多个大脑结构的LFP。市售微电极通常缺乏灵活性，不允许记录来自多个大脑结构。我们的微电极设计允许轻松修改结构以适应任何所需的结构。首先取直径50微米的dianal涂层镍铬线。将导线的一端用胶带粘在平台的背面，然后将导线缠绕在平台上最近的旋钮上三次。将导线拉伸到最远的第二个旋钮周围，在旋钮之间形成两个环。将导线在第一个旋钮上再缠绕三次，以将导线固定到位，并用胶带将导线末端固定在平台背面。将张力棒放在电线下方，并用胶带缠绕在电线上。使用显微镜和细镊子，在导线之间形成三毫米的间隙以制造皮质，腹侧，丘脑核微电极，或4.5毫米的间隙以制造纹状体 - 尼德拉微电极。如果在显微镜上使用放大倍率，请确保计算并调整放大倍率的差异和导线之间的实际距离。切割四小块 0.5 毫米厚的塑料，宽约 6 毫米，高约 3 毫米。在塑料上涂上胶水。将塑料件放在距离电线中间约一厘米的地方，距离张力棒一厘米。用棉签去除多余的胶水。胶水干燥后，用细剪刀切割电线。使用市售套件切割四个七毫米玻璃管，并将电极丝插入玻璃管中。将胶水放在玻璃管的底部，将它们连接到塑料上。等到胶水变干，然后用手术刀切割玻璃管和电线。使用超级胶水按目标区域的所需顺序粘附塑料。在塑料周围涂上环氧树脂，将电极粘合在一起。取一根粗线，在一端做一个环，将环浸入环氧溶液中，然后将其放在塑料上，确保粗线平放，以便该线可以用作手柄。将电线切割成两厘米。将电线分组，并用手术刀刮掉一毫米的隔离端。弯曲皮质电极并分离导线。使用精细的镊子，在每根导线的末端做一个环。拿着带有止血器的 10 针耳机，并使用棉签的木端在针脚上施加最少量的助焊剂。使用棉签的木端，将助焊剂涂在线环上。将电线环路焊接到 10 针耳机上。干燥耳机以防止针脚之间短路。取约0.05至0.008英寸的细线作为参考，并取地线，并从一端剥离塑料。在导线的另一端做一个循环。将基准电压源和地线的剥离侧焊接到其各自的引脚上。握住粗线，在微电极周围涂上颅骨成形水泥，特别是在电线连接到针脚的地方。避免将实际的电极末端与水泥接触。水泥干燥后，将环氧树脂放在玻璃管的底部，纹状体微电极丝以及整个电极周围。避免用环氧树脂接触电极末端。从右侧运动前皮层，左侧VL，纹状体和SNR进行局部场电位记录。癫痫发作开始被确定为电压迹线的偏转至少是基线的两倍。功率谱图显示记录的局部电位记录的频率分布。可以以毫秒级精度比较每个结构之间的癫痫发作潜伏期。在记录结束时施加电流脉冲以标记和确认形成病变的电极尖端的位置。如果微电极是为不同的大脑结构构建的，请记住根据所需结构的立体定向坐标修改导线之间的间隙。将深层结构电极放入玻璃管中，并调整微电极的长度。然后，按所需顺序连接塑料部件。

Research

JoVE Journal

Neuroscience

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

Single-unit In vivo Recordings from the Optic Chiasm of Rat

Single-unit In vivo Recordings from the Optic Chiasm of Rat

单机在体内</em从大鼠视交叉>录音

Information about the visual world is transmitted to the brain in sequences of action potentials in retinal ganglion cell axons that make up the optic ...

科研

神经科学

Functional Neuroimaging Using Ultrasonic Blood-brain Barrier Disruption and Manganese-enhanced MRI

使用超声波血脑屏障破坏和锰增强MRI的脑功能成像

Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains ...

Design and Assembly of an Ultra-light Motorized Microdrive for Chronic Neural Recordings in Small Animals

设计和组装的超轻型电动微型硬盘（Microdrive）慢性神经录音的小动物

The ability to chronically record from populations of neurons in freely behaving animals has proven an invaluable tool for dissecting the function of ...

Paired Whole Cell Recordings in Organotypic Hippocampal Slices

在器官型海马脑片搭配全细胞记录

Pair recordings involve simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling not only direct ...

Electrophysiological Method for Whole-cell Voltage Clamp Recordings from Drosophila Photoreceptors

Electrophysiological Method for Whole-cell Voltage Clamp Recordings from Drosophila Photoreceptors

全细胞电压钳记录的电生理方法<em&gt;果蝇</em&gt;光感受器

Whole-cell voltage clamp recordings from Drosophila melanogaster photoreceptors have revolutionized the field of invertebrate visual transduction, ...

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats

急性<em&gt;体内</em&gt;麻醉大鼠超导途径局部场电位和多单位活动的电生理记录

Converging evidence shows that many neuropsychiatric diseases should be understood as disorders of large-scale neuronal networks. To better understand the ...

Concurrent Recording of Co-localized Electroencephalography and Local Field Potential in Rodent

啮齿动物联合局部脑电图和局部电位的同步记录

Although electroencephalography (EEG) is widely used as a non-invasive technique for recording neural activities of the brain, our understanding of the ...

Simultaneous Recordings of Cortical Local Field Potentials, Electrocardiogram, Electromyogram, and Breathing Rhythm from a Freely Moving Rat

自由运动大鼠皮质局部电位、心电图、肌电图和呼吸节律的同步记录

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls ...

In Vivo Electrophysiological Measurement of Compound Muscle Action Potential from the Forelimbs in Mouse Models of Motor Neuron Degeneration

In Vivo Electrophysiological Measurement of Compound Muscle Action Potential from the Forelimbs in Mouse Models of Motor Neuron Degeneration

体内运动神经元变性小鼠前肢肌作用电位的电生理测量

Assessing the functionality of the nerve axon provides detailed information on the progression of neuromuscular disorders. Electrophysiological recordings ...

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

小鼠二次运动皮质中双边局部场电位记录半球横向化评价

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

构建局部场势微电极，用于同时从多个脑结构 进行体内 记录

构建局部场势微电极，用于同时从多个脑结构进行体内记录