Name: brain mapping using a graphene electrode array
Uploaded: 2023-10-20T14:45:00
Description: Watch this Scientific Journal Video about Brain Mapping Using a Graphene Electrode Array at JoVE.com

这项工作得到了仁川国立大学（国际合作）对Sunggu Yang的支持。

所有动物处理程序均已获得仁川国立大学机构动物护理和使用委员会的批准（INU-ANIM-2017-08）。
1. 动物手术准备
注意：使用Sprague Dawley Rat（8-10周大）进行本实验，没有性别偏见。
<ol><li>腹膜内用90mg / kg氯胺酮和10mg / kg甲苯噻嗪混合物麻醉大鼠。为了在整个手术过程中保持所需的麻醉深度，当大鼠出现醒来迹象时，提供补充 45 mg/kg 氯胺酮和 5...

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所提出的协议提供了一个深入的分步过程，解释了如何使用石墨烯电极阵列访问和映射大鼠的体感反应。协议获取的数据是标准必要专利，提供与每个身体部位突触相关的躯体感觉信息。
应考虑该协议的几个方面。在提取脑脊液以防止脑水肿和减轻炎症时，实验者不要损坏位于大池前面的脑干至关重要。
面部胡须提供有关周围环境的触觉感官信息，例如...

皮质图是一组局部斑块，表示大脑皮层中对感觉运动刺激的反应属性。它们是神经网络的空间形成，能够预测感知和认知。因此，皮质图可用于评估对外部刺激的神经反应和处理感觉运动信息1,2,3,4。有创和非侵入性方法可用于皮质映射。最常见的侵入性方法之一是使用皮质内（或穿透）电极来映射5,6,7,8。
使用穿透电极评估按需高分辨率皮质图面临着几个障碍。该方法太费力，无法获得像样的地图，而且侵入性太强，无法在临床上使用，从而阻碍了进一步发展。脑电图 （EEG）、正电子发射断层扫描 （PET）、脑磁图 （MEG） 和功能性磁共振成像 （fMRI） 等最新技术越来越受欢迎，因为这些技术侵入性较小且可重复。然而，鉴于其高昂的成本和解决率低，它们在有限数量的案例中使用9,10,11。近年来，具有优异信号可靠性的柔性表面电极引起了广泛的关注。基于石墨烯的表面电极表现出长期的生物相容性和机械灵活性，在复杂的大脑中提供稳定的记录12,13,14,15,16。我们小组最近开发了一种基于石墨烯的多通道阵列，用于皮质表面的高分辨率记录和位点特异性神经刺激。这项技术使我们能够长时间跟踪感官信息的皮层表征。
本文介绍了使用 30 通道石墨烯多电极阵列获取体感皮层大脑图所涉及的步骤。为了测量大脑活动，将石墨烯电极阵列放置在皮层的硬膜下区域，同时用木棍刺激前爪，前肢，后爪，后肢，躯干和胡须。记录体感区域的体感诱发电位 （SEP）。该协议也可以应用于其他大脑区域，例如听觉，视觉和运动皮层。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1mL syringe</td>
			<td>KOREAVACCINE CORPORATION</td>
			<td></td>
			<td>injecting the drug for anesthesia&#160;</td>
		</tr>
		<tr>
			<td>3mL syringe</td>
			<td>KOREAVACCINE CORPORATION</td>
			<td></td>
			<td>injecting the drug for anesthesia&#160;</td>
		</tr>
		<tr>
			<td>Bone rongeur</td>
			<td>Fine Science Tools</td>
			<td>16220-14</td>
			<td>remove the skull</td>
		</tr>
		<tr>
			<td>connector</td>
			<td>Gbrain</td>
			<td></td>
			<td>Connect graphene electrode to headstage</td>
		</tr>
		<tr>
			<td>drill</td>
			<td>FALCON tool</td>
			<td></td>
			<td>grind the skull</td>
		</tr>
		<tr>
			<td>drill bits</td>
			<td>Osstem implant</td>
			<td></td>
			<td>grind the skull</td>
		</tr>
		<tr>
			<td>Graefe iris forceps slightly curved serrated</td>
			<td>vubu</td>
			<td>vudu-02-73010</td>
			<td>remove the tissue from the skull or hold wiper</td>
		</tr>
		<tr>
			<td>graphene multielectrode array</td>
			<td>Gbrain</td>
			<td></td>
			<td>records signals from neuron</td>
		</tr>
		<tr>
			<td>isoflurane</td>
			<td>Hana Pharm Corporation</td>
			<td></td>
			<td>sacrifce the subject</td>
		</tr>
		<tr>
			<td>ketamine</td>
			<td>yuhan corporation</td>
			<td></td>
			<td>used for anesthesia</td>
		</tr>
		<tr>
			<td>lidocaine(2%)</td>
			<td>Daihan pharmaceutical&#160;</td>
			<td></td>
			<td>local anesthetic</td>
		</tr>
		<tr>
			<td>Matlab R2021b</td>
			<td>Mathworks</td>
			<td></td>
			<td>Data analysis Software</td>
		</tr>
		<tr>
			<td>mosquito hemostats</td>
			<td>Fine Science Tools</td>
			<td>91309-12</td>
			<td>fasten the scalp</td>
		</tr>
		<tr>
			<td>ointment</td>
			<td>Alcon</td>
			<td></td>
			<td>prevent eye from drying out&#160;</td>
		</tr>
		<tr>
			<td>povidone</td>
			<td>Green Pharmaceutical corporation</td>
			<td></td>
			<td>disinfect the incision area</td>
		</tr>
		<tr>
			<td>RHS 32ch Stim/Record headstage</td>
			<td>intan technologies</td>
			<td>M4032</td>
			<td>connect connector to interface cable and contain intan RHS stim/amplifier chip</td>
		</tr>
		<tr>
			<td>RHS 6-ft (1.8m) Stim SPI interface cable</td>
			<td>intan technologies</td>
			<td>M3206</td>
			<td>connect graphene electrode to headstage</td>
		</tr>
		<tr>
			<td>RHS Stim/Recording controller software</td>
			<td>intan technologies</td>
			<td></td>
			<td>Data Acquisition Software</td>
		</tr>
		<tr>
			<td>RHS stimulation/ Recording controller</td>
			<td>intan technologies</td>
			<td>M4200</td>
			<td></td>
		</tr>
		<tr>
			<td>saline</td>
			<td>JW Pharmaceutical</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>scalpel</td>
			<td>Hammacher</td>
			<td>HSB 805-03</td>
			<td></td>
		</tr>
		<tr>
			<td>stereotaxic instrument</td>
			<td>stoelting</td>
			<td></td>
			<td>fasten the subject</td>
		</tr>
		<tr>
			<td>sterile Hypodermic Needle</td>
			<td>KOREAVACCINE CORPORATION</td>
			<td></td>
			<td>remove the dura mater</td>
		</tr>
		<tr>
			<td>Steven Iris Tissue Forceps</td>
			<td>KASCO</td>
			<td>50-2026</td>
			<td>remove the dura mater</td>
		</tr>
		<tr>
			<td>surgical blade no.11</td>
			<td>FEATHER</td>
			<td></td>
			<td>inscise the scalp</td>
		</tr>
		<tr>
			<td>surgical sicssors</td>
			<td>Fine Science Tools</td>
			<td>14090-09</td>
			<td>inscise the scalp and remove the dura mater</td>
		</tr>
		<tr>
			<td>wooden stick</td>
			<td></td>
			<td></td>
			<td>whisker stimulation</td>
		</tr>
		<tr>
			<td>xylazine</td>
			<td>Bayer Korea</td>
			<td></td>
			<td>used for anesthesia</td>
		</tr>
	</tbody>
</table>

brain mapping using a graphene electrode array

皮质图表示大脑皮层中对感觉运动刺激的位置依赖性神经反应的空间组织，从而能够预测生理相关行为。各种方法，如穿透电极、脑电图、正电子发射断层扫描、脑磁图和功能性磁共振成像，已被用于获得皮质图。然而，这些方法受到时空分辨率差、信噪比（SNR）低、成本高、非生物相容性或对大脑造成物理损害的限制。本研究提出了一种基于石墨烯阵列的体感映射方法，作为皮质电图的一个特点，该方法具有优异的生物相容性、高时空分辨率、理想的信噪比和最小的组织损伤，克服了以前方法的缺点。本研究证明了石墨烯电极阵列在大鼠体感映射中的可行性。所提出的方案不仅可以应用于躯体感觉皮层，还可以应用于其他皮层，例如听觉、视觉和运动皮层，为临床实施提供先进的技术。

该协议描述了石墨烯多通道阵列如何安装在大脑表面。通过获取对物理刺激的神经反应并计算反应的幅度来构建体感图。 图1 显示了该实验的示意图。
图2A 显示了石墨烯电极阵列的结构特征。电极之间有基板的通孔。这些孔有助于电极牢固地接触皮质表面（图2B）。电极与皮层的强粘附有助于以更少的噪音记...

Watch this Scientific Journal Video about Brain Mapping Using a Graphene Electrode Array at JoVE.com

Brain Mapping Using a Graphene Electrode Array

我们提出了一种基于石墨烯阵列的脑映射程序，以减少侵入性并提高时空分辨率。基于石墨烯阵列的表面电极表现出长期的生物相容性、机械灵活性和在卷曲大脑中用于大脑映射的适用性。该协议允许同时和顺序构建多种形式的感官图谱。

使用石墨烯电极阵列进行脑映射

Cortical maps represent the spatial organization of location-dependent neural responses to sensorimotor stimuli in the cerebral cortex, enabling the ...

皮质图是一组局部斑块，代表大脑皮层中对感觉运动刺激的反应特性。它可以发现神经网络的空间形成，从而为感知和认知提供预测。因此，皮质图用于研究来自外部刺激的神经反应和感觉运动信息的处理创建皮质图是侵入性和非侵入性的方法。使用皮质内电极是用于绘制大脑皮层的最常见的侵入性方法之一。由于它可以损害大脑的高分辨率，因此无法进行进一步的测量。替代的脑图绘制工具，如EEG，PET，MEG，fMRI，是非侵入性方法，允许全脑映射和重复采样。然而，一些缺点是空间时间分辨率低、时间滞后、由于未指定的调制输入引起的误差和过高的成本。钙成像和光遗传学功能磁共振成像等光学技术可提供大规模的脑映射，但由于指示剂毒性和低时空分辨率，在临床上没有益处。石墨烯电极阵列表现出长期的生物相容性和机械灵活性，可提供卷曲大脑的稳定记录。最近，我们小组开发了一种石墨烯电极阵列，可为皮质表面的神经信号提供高分辨率记录。该协议使用石墨烯电极阵列记录来自SD大鼠前爪，前肢，后爪，后肢，躯干和胡须的SEP，以创建皮质图。腹膜内注射氯胺酮和甲苯噻嗪混合物，氯胺酮每公斤90毫克，甲苯噻嗪每公斤10毫克。为了在整个手术过程中保持所需的麻醉深度，如果大鼠出现觉醒迹象，则注射每公斤 45 毫克氯胺酮和每公斤 5 毫克甲苯噻嗪鸡尾酒。捏住脚趾以确认麻醉深度。如果老鼠颤抖，再等几分钟，直到它没有反应。使用修剪器从眼睛之间的空间到耳朵后部剃掉老鼠头上的皮毛。然后将眼药膏涂在眼睛上。使用耳杆和立体定位适配器，将大鼠的头部固定在立体定位装置上。确保老鼠的头部固定正确。用聚维酮浸泡的棉花交换对剃光区域进行消毒。用酒精擦洗剃光的区域。重复灭菌过程三次。然后在头皮中注射0.1毫升2%利多卡因，诱导局部麻醉。创建一个两到三厘米的中线切口，横向拉开头皮以露出头骨。用蚊钳夹住头皮，露出头骨。首先用镊子刮擦颅骨表面来去除骨膜。在朝向λ的直线上，通过撕裂后缘枕骨后面的肌肉来定位胸骨大区。使用显微镜近距离观察大池。轻轻切开水池。撕裂大池后，脑脊液流出。用盘绕的无菌纱布排出脑脊液，使大脑下沉。根据前膛的位置，根据预定义的立体定位坐标标记石墨烯电极的放置位置。躯体感觉皮层位于右半球颅骨前侧轴 3 毫米处，右侧侧向 6 毫米处。根据立体定位坐标钻标记的区域。用骨头去除颅骨。将棉签插入 26 花哨的针头中。将针弯曲至90度。要去除硬脑膜，请使用弯曲的针创建一个孔。向上提起硬脑膜，将镊子插入该孔中，然后继续用剪刀切割硬脑膜或用镊子撕裂。将用生理盐水弄湿的无菌纱布附着在躯体感觉皮层上，以防止皮层变干。通过应用盐溶液分离石墨烯多电极阵列而不会造成任何损坏。从连接器上拆下参考线和接地线的外壳。要设置石墨烯多电极阵列以进行映射，请将头部平台与连接器连接。将石墨烯电极阵列与连接器连接。将接口电缆插入录音控制器。将石墨烯多电极阵列和连接器复合体连接到接口电缆。然后将石墨烯多电极阵列复合物固定在立体定位臂上。根据预定义的立体定位坐标将石墨烯电极阵列放置在体感皮层上。将参考线放在枕骨后面的组织中。然后将地线连接到接地的光学工作台。打开录音软件并将采样率设置为 30 千赫兹，然后单击 OK 按钮。设置 60 赫兹陷波滤波器以消除电源噪声。用细棍弯曲胡须。通过数据采集系统记录所需时间的神经信号。这些信号是通过对胡须的刺激获得的神经信号。其他身体部位也以同样的方式刺激记录标准必要专利。打开名为 read_Intan_RHS2000 文件的 MATLAB 代码。单击运行按钮。选择录制文件并单击该文件，然后等待文件被读取。输入命令图以创建 2D 线图。选择响应刺激的代表性峰值信号，并通过测量最大值和最小值来计算SEP的幅度。根据SEP的振幅，用不同的颜色阴影对等级进行着色，从而获得地形图。石墨烯电极阵列在左边。所有电极之间都有基板的通孔。这些孔有助于电极与皮层保持牢固的接触。在右边，图像显示了皮层上的电极阵列。在左边，由刺激引起的每个位置依赖性神经反应对大鼠身体不同部位的比率以大鼠的形状显示。每个身体部位都用不同的颜色表示。这将配置躯体感觉皮层图。在右图上，下图显示了通过放置在皮层表面的多通道石墨烯电极获得的刺激特异性反应。每个彩色框代表不同身体部位在 30 个通道上的响应。上图表示使用石墨烯电极阵列在体感皮层表面检测到的LFP。颜色条显示较大的振幅由较深的颜色阴影表示，每种颜色表示不同的身体部位。该图根据颜色条中的阴影弯曲振幅。啮齿动物的这个人类是使用刚才解释的结果创建的。某个身体部位的振幅越大，它在同胞体上的显示就越大。下图代表了从每个身体部位收集的LFP的单独数据。它表示基于颜色条检测到的LFP的不同振幅。此过程有几种预防措施。在去除脑脊液时，实验者应注意不要触摸脑干或脊髓。这一步需要练习。啮齿动物的胡须发育到足以感知偏转方向、刺激强度和被刺激的胡须的位置。这就是为什么该协议应以恒定的方向和强度刺激晶须的原因。其他身体部位也应持续刺激。该协议的局限性在于，当石墨烯电极阵列安装在皮质表面上时，无法记录深部大脑结构中诱发的信号。因此，实验者无法确定列式网络是如何在神经反应方面分层组织的。尽管如此，如果实验者改变放置电极阵列的多个区域，则可以进一步应用该协议。例如，如果实验者将电极放在听觉或视觉皮层上以提取听觉和视觉信息，他们可以获得听觉或视觉图。此外，该协议可以扩展到石墨烯多电极阵列的慢性植入。

Research

JoVE Journal

Neuroscience

Multi-electrode Array Recordings of Neuronal Avalanches in Organotypic Cultures

器官型培养的神经元雪崩多电极阵列记录

The cortex is spontaneously active, even in the absence of any particular input or motor output.  During development, this activity is important for the ...

科研

神经科学

Mapping Cortical Dynamics Using Simultaneous MEG/EEG and Anatomically-constrained Minimum-norm Estimates: an Auditory Attention Example

同时MEG / EEG和解剖约束的最小模估计皮质动态映射：听觉注意示例

Magneto- and electroencephalography (MEG/EEG) are neuroimaging techniques that provide a high temporal resolution particularly suitable to investigate the ...

A Computer-assisted Multi-electrode Patch-clamp System

计算机辅助多电极膜片钳系统

The patch-clamp technique is today the most  well-established method for recording electrical activity from individual  neurons or their subcellular ...

Functional Imaging of Auditory Cortex in Adult Cats using High-field fMRI

听觉皮层在成年猫用高场磁共振成像功能成像

Current knowledge of sensory processing in the mammalian auditory system is mainly derived from electrophysiological studies in a variety of animal ...

Neural Activity Propagation in an Unfolded Hippocampal Preparation with a Penetrating Micro-electrode Array

神经活动的传播的展开海马准备具有穿透微电极阵列

This protocol describes a method for preparing a new in vitro flat hippocampus preparation combined with a micro-machined array to map neural activity in ...

Direct-current Stimulation and Multi-electrode Array Recording of Seizure-like Activity in Mice Brain Slice Preparation

小鼠脑切片准备扣押样活性的直接电流刺激和多电极阵列记录

Cathodal transcranial direct-current stimulation (tDCS) induces suppressive effects on drug-resistant seizures. To perform effective actions, the ...

Adaptation of Microelectrode Array Technology for the Study of Anesthesia-induced Neurotoxicity in the Intact Piglet Brain

微电极阵列技术在完整仔猪脑麻醉诱导神经毒性研究中的应用

Every year, millions of children undergo anesthesia for a multitude of procedures. However, studies in both animals and humans have called into question ...

Chronic Implantation of Whole-cortical Electrocorticographic Array in the Common Marmoset

常见马莫塞的全皮质电皮质阵列的慢性植入

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent ...

Translational Brain Mapping at the University of Rochester Medical Center: Preserving the Mind Through Personalized Brain Mapping

罗切斯特大学医学中心的转化脑图:通过个性化大脑映射保护大脑

The Translational Brain Mapping Program at the University of Rochester is an interdisciplinary effort that integrates cognitive science, neurophysiology, ...

Real-Time fMRI Brain Mapping in Animals

动物实时功能磁共振成像大脑映射

Dynamic fMRI responses vary largely according to the physiological conditions of animals either under anesthesia or in awake states. We developed a ...