Name: concurrent recording of co-localized electroencephalography and local field potential in rodent
Uploaded: 2017-11-30T12:30:00.000+00:00
Duration: 8 min 31 s
Description: Watch this Scientific Journal Video about Concurrent Recording of Co-localized Electroencephalography and Local Field Potential in Rodent at JoVE.com

我们要感谢安德鲁 Cripps 和雷丁大学的资源单位。这项研究由 BBSRC (格兰特号: BB/K010123/1) 资助。与这项工作相关的数据可从 Y.Z. (ying.zheng@reading.ac.uk 英 zheng@reading) 免费获得。

所有的实验都是按照英国内政部 (《动物 (科学规程) 法》, 1986) 进行的, 并由英国雷丁大学的研究伦理委员会批准。
1. 动物制剂
注意: 所有的实验都使用了女性李斯特的大鼠。这是一个非生存的程序。
<ol><li>把老鼠的体重记录在实验室的秤上。</li>
	<li>麻醉5% 异氟醚和1升/分的氧流量。</li>
	<li>把老鼠放在一个立体定位器上, 用纸巾在它的身体下面, 用它的牙齿通过咬杆休息。纸巾将使热垫的插入更容易 (见步骤 2.3) 和捕捉任何粪便从老鼠在实验过程中。</li>
	<li>通过一个硬塑料鼻锥安装在鼻子钳上连续管理异氟醚, 在3% 的浓度为0.5 升/分钟的氧气流量的大鼠适配器. 将锥体连接到小动物异氟醚麻醉系统。</li>
</ol>2. 手术程序
<ol><li>在大鼠休息的纸巾下面插入一个恒温加热垫, 用两个耳条固定鼠头, 用直肠温度计监测体温。</li>
	<li>剃掉老鼠头上的毛</li>
	<li>将眼部软膏涂抹于眼部, 防止角膜干燥。</li>
	<li>在暴露颅骨前, 应将利多卡因滴到头皮上, 轻轻按摩至皮肤。</li>
	<li>在头皮上做一个大约2-3 厘米的中线切口, 用手术刀来露出颅骨的表面。</li>
	<li>小心地将颞肌肉对侧向的触须垫用 Jacquette 器和一对锯齿和弯曲的解剖钳从颅骨刺激。必要时用棉签清洁颅骨。</li>
	<li>用编织的丝绸, 不可吸收的缝合, 栓分开的肌肉对头皮与紧结, 然后栓缝合牢固地对立体定向框架18。</li>
	<li>使用立体定向坐标定位的桶皮层, 2.5 毫米尾管到 bregma 和6毫米侧向中线19。用铅笔或记号笔在体感皮层的位置画一个点。</li>
	<li>
用牙科钻头在标记的位置钻一个毛刺孔。为防止颅骨在钻孔过程中过热, 应将无菌生理盐水 (氯化钠 0.9%) 应用于工作区每10-15 秒。钻探过程包括以下3步骤:
	<ol>
<li>钻一个直径 < 2 毫米孔到头骨使用钻头 #4 (0.055 直径)。当心不要钻到硬脑膜上。</li>
		<li>用钻头 #1/4 (直径 0.019) 将孔的底部变薄至半透明。</li>
		<li>使用27克针刺穿硬脑膜, 允许插入微电极。</li>
	</ol>
</li>
	<li>将大鼠, 固定在一个立体定向的框架上, 转移到一个安装在隔振工作站顶部的法拉第笼上。</li>
	<li>将一个血传感器钳连接到血控制单元, 对鼠的后爪进行连续监测以下生理参数: 心率、呼吸率、动脉氧饱和度、脉搏扩张和呼吸扩张。这些参数在 PC 显示器上连续显示, 反映了大鼠的生理状况和麻醉深度。</li>
	<li>更换硬塑料鼻锥的异氟醚管理和鼻钳的大鼠适配器与 microflex 呼吸器装有一个透明的软鼻子锥, 这是修改 (图 1A), 使容易晶须刺激的一侧的晶须垫不损害异氟醚的管理。</li>
	<li>插入两个不锈钢刺激电极到晶须垫暴露的切出在鼻子锥。</li>
	<li>将刺激电极连接到一个孤立的电流刺激器。</li>
	<li>用镊子提起颈部中线的皮肤, 用剪刀作 1 ~ 2 厘米切口, 准备放置参考电极。注意不要切断肌肉组织。</li>
</ol>3. Co 局部脑电图/LFP 设置
<ol><li>用棉签清洁和干燥毛刺孔周围的头骨。</li>
	<li>小心地将传导性脑电图膏放在 eeg 蜘蛛电极的一个扁平一侧。在蜘蛛电极上留下一个小洞, 清除脑电波膏, 允许多层微电极穿过孔而不接触粘贴和蜘蛛电极。这可以防止 EEG 电极和微电极之间的电接触。</li>
	<li>将蜘蛛电极与头骨上的毛刺孔对齐, 并将脑电波糊朝向头骨。</li>
	<li>小心地将蜘蛛电极压在头骨上, 通过脑电图膏与头骨进行牢固接触。用注射器上的针头将毛刺孔上的任何糊状物去除。</li>
	<li>除去蜘蛛电极周围的过多的脑电波膏, 使蜘蛛电极和头骨之间的接触空间被限制在电极的大小 (图 1B)。</li>
</ol><img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56447/56447fig1.jpg"> 图 1: 并发脑电图/LFP 记录的常规设置.(A) 设置由一个改进的鼻锥, 以便于在异氟醚麻醉下的晶须垫刺激, 两个刺激电极插入晶须垫, 一个蜘蛛电极定位在头骨上方的桶皮层对侧刺激电极, 通过蜘蛛电极插入到桶皮层的多通道微电极, 以及放置在老鼠颈部后部切口内的参考电极。(B) 通过显微镜将蜘蛛电极安全地放置在头骨上的脑电图贴。微电极入到在蜘蛛电极下钻入头骨的毛刺孔中。头皮被绑在立体定向框架上的外科手术线 (缝合) 来控制。<a href="//cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/56447/56447fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<ol start="6"><li>将脑电图膏涂抹在参考电极上, 并将其安全地放在大鼠颈部后部的切口内。</li>
	<li>通过一个低阻抗信号的被动信号分配器将 EEG 电极连接到前置放大器 (图 2)。确保蜘蛛电极的阻抗低于5ω。如果不是, 检查 eeg 膏是否与颅骨有良好的接触, 并将电极牢牢地压在 eeg 糊上。如有必要, 添加更多脑电图膏。</li>
	<li>在立体定向架上安装一微臂。连接一个线性16通道微电极 (100 µm 间距, 每个站点177µm2) 到16通道的急性 headstage, 将安全地夹在微臂上。</li>
	<li>在脑电图和微电极上涂抹脑电图贴在参考电极上, 然后将它们安全地放置在切口内 (图 1A)。</li>
	<li>调整微臂的角度, 使微电极与皮质表面垂直。这个角度通常在25-35 °之间。</li>
	<li>在显微镜下通过转动微旋钮降低微电极, 使电极尖端对准毛刺孔底部的微小开口, 直到最上部的电极穿透皮质表面。必须注意避免将微电极强制到硬脑膜表面, 因为这会破坏电极。</li>
	<li>将16通道微电极与通过光纤电缆连接到数据采集单元的前置放大器耦合 (图 2)。</li>
	<li>打开前置放大器、数据采集单元和连接到设备的计算机。打开刺激盒。</li>
	<li>通过缓慢地将微的 z 轴旋钮旋转到1500µm20的深度, 将微电极正常插入皮质表面。</li>
	<li>微调整的深度, 应用一列车刺激的晶须垫和观察16通道诱发 LFP 在 pc 显示器上使用的软件的数据采集单元安装在 pc 上。小心地转动 z 轴旋钮在微直到最高振幅的诱发 LFP 发生在通道7附近 (因为这与第四层在皮层)。 注: 信息学侧脑电电极设置: 对于一些实验, 第二个蜘蛛电极被放置在信息学侧侧的完整头骨上方的桶皮层。该装置允许在静止状态下进行双侧脑电图记录, 以研究毛刺孔对 eeg 信号的影响。 注: 建立脑电图电极的手术方法与上述相同, 但在步骤2.6 中, 头部两侧的颞肌肉小心地与颅骨分离, 缝合背部并牢固地绑在相应的一侧立体定位框架。 注: 同期脑电图/LFP 设置也与上述描述相同, 与一个额外的步骤, 第二个蜘蛛电极加载脑电图糊, 然后紧紧压在头骨上方的信息学侧筒皮层。</li>
</ol><img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/56447/56447fig2.jpg"> 图2。一个信号流图.老鼠被放在法拉第笼子里。刺激电极通过其安装在 PC 上的软件, 接收由数据采集单元控制的刺激器盒中的命令。将微电极记录的神经信号传送到法拉第笼内的预放大器。EEG 探针记录的神经信号通过信号分配器传输到前置放大器。预放大器通过光纤电缆连接到法拉第笼外的数据采集单元。然后将神经数据存储在 pc 上的本地驱动器上, 同时它们也可以显示在 pc 显示器上。一种移动式小动物异氟醚系统从法拉第笼外管理异氟醚。<a href="//cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/56447/56447fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
4. 电刺激和神经记录
注: 所有神经数据的采样频率为 24.41 kHz, 具有16位分辨率。试验包括在试验开始时的一次电刺激。每项试验持续十年代, 这也是间刺激间隔 (ISI)。每一个刺激都是一个1.2 毫安持续0.3 毫秒的方波电流脉冲用于研究毛刺孔的作用, 并记录250s 的连续休眠状态。
<ol><li>打开正在使用的计算机上的录制软件。</li>
	<li>从 "OpenProject" 的下拉菜单中选择 "加载项目...", 为实验加载正确的电路。将出现一个新窗口 ("工作台") (图 3)。</li>
</ol><img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/56447/56447fig3.jpg"> 图3。数据获取单元的软件 GUI 的显示.它允许上传合适的电路, 设置刺激参数, 记录和可视化数据。<a href="//cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/56447/56447fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<ol start="3"><li>
创建一个新的目录 (称为 "坦克" 的软件) 存储神经录音。
	<ol>
<li>单击窗口顶部的 "文件", 然后选择 "数据任务管理"。将出现一个新窗口 ("坦克管理")。</li>
		<li>在 "容器管理" 窗口中, 按鼠标右键以显示菜单。选择 "创建新坦克"。将出现另一个新窗口 ("创建数据池")。</li>
		<li>在 "创建数据池" 窗口中, 选择计划创建新数据目录的路径, 然后输入新目录的名称。然后按 "确定"。此窗口将消失。</li>
		<li>新目录将出现在 ' 坦克管理 ' 窗口, 但在灰色。通过右键单击注册此目录, 然后从下拉菜单中选择 "注册罐"。新目录的名称左侧将出现一个红色的星号和一个绿色箭头 (图 4)。</li>
		<li>通过右键单击 "容器管理" 窗口并在下拉菜单中选择 "刷新罐列表", 注销未使用的任何以前的目录。</li>
		<li>单击 "确定" 退出 "容器管理" 窗口。</li>
	</ol>
</li>
</ol><img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/56447/56447fig4.jpg"> 图 4: 显示已注册数据目录的软件 GUI.<a href="//cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/56447/56447fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<ol start="4"><li>
在 "范围" 中注册新目录, 在实验中显示神经信号。
	<ol>
<li>单击 "OpenProject" 窗口中的 "范围" 图标。将出现一个新窗口 ("范围")。</li>
		<li>右键单击 "范围" 窗口中的鼠标, 然后在下拉菜单中选择 "刷新罐列表"。新目录的名称将以灰色显示。</li>
		<li>单击新目录。红色星和绿色箭头将出现在新目录的名称左边, 现在是黑色的。</li>
	</ol>
</li>
	<li>通过单击窗口顶部的 "安装", 在 "工作台" 窗口中设置数据获取的实验参数。将出现一个新窗口。选择 "扫描回路", 设置试验的长度和要记录的试验数。</li>
	<li>检查刺激框是否打开。</li>
	<li>按 "工作台" 窗口中的 "记录" 按钮。将出现一个新窗口。输入要为实验运行保存的数据文件的名称, 但在这个阶段不按返回按钮, 因为需要设置 EEG 记录参数。</li>
	<li>
使用预放大器上的图形用户界面 (GUI) 设置 EEG 记录参数。触摸屏幕 (任何地方) 的前置放大器唤醒屏幕。选择 "解锁" 以解锁显示 (图 5)。
	<ol>
<li>在 "2: 脑电图" 面板中按左图标。将显示新的显示。</li>
		<li>按 ' 耦合 ', 并选择 ' AC '。</li>
		<li>按 "参考模式" 并选择 "本地"。</li>
		<li>按 "桑普率", 然后选择 "25KHz"。</li>
		<li>按 "确定" 返回到原始显示。</li>
	</ol>
</li>
</ol><img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/56447/56447fig5.jpg"> 图 5: 预放大器上的 GUI.它允许设置 EEG 记录参数 (例如、采样频率和引用首选项)。<a href="//cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/56447/56447fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<ol start="9"><li>通过按下 "2: 脑电图" 面板中的中间图标, 检查脑电图探头的阻抗。如果太高, 在探头上添加更多的脑电图膏。按 "确定" 返回到原始显示。</li>
	<li>等待二十年代, 以避免记录脑电图记录的初始波动。</li>
	<li>返回到 PC 监视器 (在二十年代等待之后), 然后按键盘上的 "返回" 键。脑电图和 LFP 信号将被记录。</li>
</ol>5. 数据分析
<ol><li>
前处理的诱发 LFP 和脑电图信号的试验的基础上, 使用以下步骤。
	<ol>
<li>通过20样本 (相当于0.82 毫秒) 将神经数据及时移回。这是用于收集 TDT 本身的神经数据的电路所产生的延迟。通过转移数据, 时间零点与刺激的开始是一致的。</li>
		<li>移除刺激伪影, 将神经数据从0到1毫秒, 用一条直线连接数据点在0毫秒, 数据点在1毫秒。</li>
		<li>零意味着每一个试验减去的平均值的神经信号200毫秒前刺激开始。</li>
		<li>低通滤波器在800赫兹以下的数据使用4的th命令, 在两个方向上都有一个类型过滤器, 以避免在数据中引入任何时间偏移。</li>
		<li>对齐跨动物的多层流数据。对于每个动物的 LFP 数据, 应用逆电流源密度 (样条中心, 源半径 R = 0.5 mm) 分析21与高斯滤镜 (λ = 50 µm), 以找到第四层下沉1, 这是由最大的负峰值发生在皮质深度低于脑膜表面在前15毫秒的刺激开始。CSD 和相应的 LFP, 然后根据它们在动物中的下沉位置对数据进行排列。共水槽位于 IV 层, ~ 600 µm 以下的脑膜表面。</li>
		<li>对齐后, 使用通道2、7和12的重新调整的 LFP 作为代表的神经反应的 supragranular, 颗粒状, 和 infragranular 层, 分别在桶皮层。</li>
	</ol>
</li>
	<li>通过平均100试验前处理数据, 计算平均诱发 LFP 和脑电图。</li>
	<li>为了研究毛刺孔对 eeg 的影响, 将 eeg 信号下采样到1000赫兹, 并计算出在250s 周期内侧向侧 (有一个孔在颅骨中) 和信息学侧面 (完好的头骨) 蜘蛛电极记录的功率谱密度 (PSD)休息状态。PSD 是从 0.1-100 赫兹的 Matlab 使用的函数 "pmtm", 这是基于多方法22。</li>
	<li>将频率范围划分为以下已知的频带: 三角洲 (δ): 0.1-4 赫兹, θ (): 4-8 赫兹, α (α): 8-13 hz, β (β): 13-31 hz, 伽玛 (γ): 31-100 赫兹. 计算每个波段内的平均 PSD。</li>
	<li>在每个频带内, 使用该公式计算出在信息学侧脑电波之间, Perr之间的归一化差异: <img alt="Equation" src="/files/ftp_upload/56447/56447eq1.jpg"> 其中, Pc 和pi 是信息学侧脑电图的平均 PSD, 分别在感兴趣的频率波段。</li>
	<li>在每个频带内, 执行一个样本 t 检验, 以测试在两个半球记录的 EEG 信号 PSD 之间没有显著差异 (在0.05 的意义级别) 的假设。</li>
</ol>

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作者没有什么可透露的。

我们描述了一个实验过程, 同时记录的 co 局部脑电图和 LFP 信号的异氟醚麻醉大鼠响应晶须垫刺激。一个微电极入到大脑皮层通过一个开放的脑电图蜘蛛电极 , 这是与一个毛刺孔钻入头骨。电极通过导电性和粘附脑电浆23固定在颅骨上。用于管理异氟醚的鼻锥被修改, 使刺激电极可以轻松地插入晶须垫。
EEG 膏是有效的安装蜘蛛电极安全地对头骨, 同时提供优良的电导率在整个实验日不需要额外的应用膏。它取代了不可取的使用胶水固定的蜘蛛电极的周围的头骨, 因为胶水是不导电的, 可以增加阻抗的电极, 如果它运行之间的头骨和电极。eeg 糊在脑电凝胶上有许多优点, 在毛刺孔周围不易形成, 在实验过程中会产生干燥, 导致脑电信号不佳。
当鼠被放置在法拉第笼内时, 由于环境的电噪声大大减弱。然而, 有时神经信号仍然相当嘈杂。在大多数情况下, 这是由于参考电极没有安全定位, 因此需要重新调整或更多的脑电图粘贴使用。另一个常见的问题是诱发 LFP 的振幅较小。这可能是由于微电极没有定位在皮层区中心激活的刺激电极。而不是重新插入微电极, 这可能会对局部神经元造成更多的损害, 我们通常调整的刺激电极在晶须垫的位置, 直到一个合理的幅度的 LFP (> 3 mV) 可以观察到。
该技术的局限性之一是蜘蛛电极的空间分辨率较差, 其直径为 6 mm。这与大鼠头骨的大小相比是很大的。不幸的是, 这里使用的蜘蛛电极是最小的可供购买的。将蜘蛛电极的直径减小到2-4 毫米是可取的, 从而增加了 eeg 记录的空间特异性, 使得 eeg 信号与 supragranular LFP 信号的比较不明确。
协议中的几个关键步骤需要特别注意。首先是通过毛刺孔插入微电极。由于硬脑膜是其他完整的, 插入的精度是至关重要的。电极尖端的轻微电阻通常意味着电极没有正确定位。它必须提出, 位置调整, 并重新插入。第二个是鼻子锥在老鼠身上的位置。它不能太松散, 因为异氟醚将从锥中逃脱。它也不能太紧, 因为这会阻碍老鼠的鼻孔, 造成呼吸困难。还需要特别注意, 以确保 EEG 记录的振幅比 LFP 的顶声道记录小得多 (通常小于5到10倍)。如果它们是相似的, 这是一个迹象表明, EEG 探针已进入直接或间接接触的微电极。间接接触通常是通过脑脊髓液 (CSF), 有时填补孔钻在头骨。CSF 的电导率通常是颅骨24,25的100倍。因此, 如果在毛刺孔内的脑脊液水平足够高, 它可以与蜘蛛电极接触。为了避免这种, 应该经常清洗的孔与超级吸水性棉海绵, 如吸收矛。
通过将另一只蜘蛛电极放在信息学侧筒皮层上的完整颅骨上, 研究了颅骨上的毛刺孔 (直径 < 2 mm) 对脑电记录的影响, 从而可以比较双边脑电图记录。在图 9 和图 10中显示的结果表明, 在0.05 级意义上的效果无关紧要。影响 eeg 振幅的其他因素包括脑电膏与颅骨接触的程度, 电极被压到糊状的方式, 以及脑电波膏在颅骨上的空间范围。
同样值得注意的是, 这里所描述的协议记录了颅内脑电图, 这与人类脑电图研究中使用的头皮脑电图不同。头皮的作用就像一个电阻器或一个低通滤波器, 它将进一步减少 EEG 记录的信噪比。
最后, 通过对 ERP 的时间动态和皮质层诱发 LFP 的比较, 表明体感诱发电位在皮层 supragranular 层中的 LFP 比颗粒状和 infragranular 层中更好。这与我们先前的工作6一致, 表明 ERP 的初始段 (P1) 与在颗粒层中发生的兴奋性突触电流的流入所产生的返回电流有关, 而随后的减少 (N1) 在 ERP 中可能与丘脑传入的延迟到达皮层层 II/III 和/或来自更深皮层层的前馈信号有关。总之, eeg/LFP 的并发记录可以增强对 eeg 的神经起源的理解, 并促进脑电信号在皮层层间的神经信号的数学建模。

一般认为, 通过电极记录的 LFPs 主要反映了局部锥体神经群的同步兴奋和抑制突触活动的加权和,1,2,3,4. 我们最近的研究表明, LFP 信号的剖面可以被分离成励磁和抑制的组成部分5,6。然而, 由于 LFP 通常是通过侵入性的程序来测量的, 所以它不适合人脑的大部分研究。
另一方面, 脑电图是一种非侵入性的测量大脑电活动的技术。它被广泛应用于某些类型的神经疾病如癫痫的诊断工具, 作为人类认知研究的一个研究工具。尽管它很受欢迎, 但 EEG 的一个主要限制是无法精确地根据基础神经信号7,89来解释其时间分布。
越来越多的, 数学模型的脑电图发展, 以提高对大脑功能的理解10,11,12,13,14,15。现有的脑电模型大多是基于模型拟合的频域特征, 在自发活动期间对 eeg 数据谱进行预测, 很少有脑电模型能产生逼真的感官诱发电位。在此背景下, eeg 和 LFP 的并发记录将为开发更精确的脑电图数学模型提供重要的洞察力和制约因素。
为了解决这一需要的并发录音进一步探索神经起源的脑电图, 我们开发了一种方法, 同时记录脑电图和多层 LFP 信号在新大脑皮层的麻醉大鼠。设置类似于以前在灵长类动物中进行的脑电图/LFP 研究16,17。我们进一步研究了一个钻孔入颅骨的毛刺孔对周围的 eeg 记录的影响, 通过比较双边脑电图记录 (即, 一个半球与一个毛刺孔, 另一个半球完好) 在缺乏感官刺激.我们的结果表明, 并发脑电图/LFP 录音可以简单和有效地进行, 很少 eeg 信号失真的毛刺孔在头骨。

<table><tbody><tr><td>Female Lister Hood rats</td><td>Charles Rivers</td><td></td><td></td></tr><tr><td>Spider electrode</td><td>Unimed Electrode Supplies Ltd</td><td>SCS24-426</td><td></td></tr><tr><td>EEG paste: Ten20</td><td>Unimed Electrode Supplies Ltd</td><td>10-20-S</td><td></td></tr><tr><td>Stereotaxic holder with dual micromanipulator arms: Dual Manipulator Stereotaxic Frame with 18&amp;deg; Ear Bars</td><td>WPI (World Precision Instruments)</td><td>502603</td><td></td></tr><tr><td>Isoflurane</td><td>National Vet Services Limited</td><td>50878</td><td></td></tr><tr><td>Hard plastic nose cone: Anasthesia Gas Mask for Rat</td><td>WPI</td><td>502054</td><td></td></tr><tr><td>Small animal isoflurane anaesthetic system</td><td>WPI</td><td>EZ-B800A</td><td></td></tr><tr><td>Thermostatic heating pad: Rat Blanket System 230V</td><td>Harvard Apparatus UK</td><td>50-7221-F</td><td></td></tr><tr><td>Ophthalmic ointment: Optixcare eye lube</td><td>Viovet</td><td>203865</td><td></td></tr><tr><td>Lidocaine Hydrochloride (Injection 2%)</td><td>Larkmead Vets</td><td></td><td></td></tr><tr><td>Jacquette Scaler #1SSE, 18cm, Hollow</td><td>WPI</td><td>503421</td><td></td></tr><tr><td>Serrated and curved dissecting forceps</td><td>WPI</td><td>15915</td><td></td></tr><tr><td>Braided silk, non-absorbable suture: Mersilk Suture W502H</td><td>National Vet Services Limited</td><td>153746</td><td></td></tr><tr><td>Dental drill: BONE MICRO DRILL SYST 230 VAC</td><td>Harvard Apparatus UK</td><td>72-4860</td><td></td></tr><tr><td>Sterile Saline: Sodium chloride 0.9%</td><td>Animalcare Ltd</td><td>14K26BT</td><td></td></tr><tr><td>Drill bit #4 : Ball Mill, Carbide, #4</td><td>Harvard Apparatus UK</td><td>72-4958</td><td></td></tr><tr><td>Drill bit #4 : Ball Mill, Carbide, #1/4</td><td>Harvard Apparatus UK</td><td>72-4962</td><td></td></tr><tr><td>Faraday cage</td><td>Newport Corporation</td><td>VIS-FDC-3600</td><td></td></tr><tr><td>Vibration isolation workstation: Vision IsoStation</td><td>Newport Corporation</td><td>M-VIS3660-RG4-325A</td><td></td></tr><tr><td>Oximeter Control Unit and sensor: MouseOxPlus, Starr Life Sciences Corp.</td><td>WPI</td><td>O15001</td><td></td></tr><tr><td>Transparent soft nose cone: Microflex Non-Rebreathing Unit with a Rat Nosecone</td><td>WPI</td><td>EZ-103A</td><td></td></tr><tr><td>Stainless steel stimulating electrodes</td><td>PlasticsOne</td><td>E363/1/SPC</td><td></td></tr><tr><td>Isolated current stimulator</td><td>Made in House</td><td></td><td></td></tr><tr><td>16-channel micro-electrode, 100 &amp;mu;m spacing, area of each site 177 &amp;mu;m&lt;sup&gt;2&lt;/sup&gt;</td><td>NeuroNexus</td><td>A1x16-10mm-100-177-A16</td><td></td></tr><tr><td>16-channel acute headstage</td><td>Tucker David Technologies Inc., TDT</td><td>RA16AC-Z</td><td></td></tr><tr><td>Pre-Amplifier: Z-Series 64-Channel Neuro-Digitizing Preamp</td><td>TDT</td><td>PZ5-64</td><td></td></tr><tr><td>Passive signal splitter: 32-Channel Splitter Box for PZ5</td><td>TDT</td><td>S-BOX_PZ5</td><td></td></tr><tr><td>Data acquisition unit: RZ2 BioAmp Processor. Z-Series 4-DSP ultra high performance processor</td><td>TDT</td><td>RZ2-4</td><td></td></tr><tr><td>Software for Neurophysiology: OpenEX</td><td>TDT</td><td></td><td></td></tr><tr><td>Matlab</td><td>MathWorks</td><td></td><td></td></tr><tr><td>Absorption spears</td><td>Fine Sicence Tools</td><td>18105-01</td><td></td></tr></tbody></table>

concurrent recording of co-localized electroencephalography and local field potential in rodent

尽管脑电图 (eeg) 被广泛应用于记录大脑神经活动的非侵入性技术, 但我们对脑电图的脑组织的认识仍然非常有限。通过多层微电极记录的局部场电位 (LFPs) 可以更详细地描述大脑皮层不同皮层层的同时神经活动, 但这种技术是侵入性的。将脑电图和 LFP 测量结合在一个前临床模型中, 可以极大地提高对脑电信号生成中所涉及的神经机制的认识, 并有助于推导出更逼真和生物学上准确的脑电图数学模型。本文介绍了一种在麻醉啮齿动物中获得并发和 co 局部脑电图和多层流 LFP 信号的简单方法。我们还调查了脑电信号是否有明显的影响, 在头骨钻孔的一个毛刺孔插入一个微电极。我们的结果表明, 毛刺孔有一个微不足道的影响脑电图记录。

4只大鼠的数据均为平均时间序列。诱发脑电反应的振幅, 也被称为事件相关电位 (ERP), 通常比 LFP 的幅度小得多。图 6分别显示了 supragranular、粒状和 infragranular 层的平均 ERP 和 LFP。每个图形中的错误带是相应的标准错误。可以看出, ERP 大约比诱发 LFP 小10倍。
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/56447/56447fig6.jpg"> 图 6: 平均 (n = 4) ERP、supragranular、粒状和 infragranular LFP 的神经信号.阴影表示标准错误。<a href="//cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/56447/56447fig6large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
对 ERP 和 LFP 的时间动态进行比较, 如图 7所示。ERP 和 supragranular LFP 在图 7A中的直接叠加说明了这两种类型的神经信号之间的振幅差的顺序。为了比较时间动态, ERP 和 LFP 都是相对于其负峰值振幅正常化。图 7B和7C分别显示规范化的 ERP 与规范化的 supragranular LFP 和规范化的粒 LFP 的叠加。
从图 7B可以看出, ERP 中的 P1 和 N1 的峰值比 supragranular 层中相应的 LFP 峰值更延迟。然而, 这两个神经信号的时间分布是相似的, 与 P1 前 N1。另一方面, ERP 的时间分布明显不同于 LFP (图 7C) 的粒度 (第四层)。重要的是, 他们不是彼此的镜像, 颗粒 LFP 由一个单一的负峰值 (反映在皮质层 IV 的主要下沉), 而 ERP 主要由两个相对极性的峰值。
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/56447/56447fig7.jpg"> 图 7: ERP 和 LFP 时间动态的比较.(A) ERP (实线) 叠加 supragranular LFP (虚线)。阴影表示标准错误。(B) 规范化的 ERP (实线), 与规范化的 supragranular LFP (虚线) 叠加。(C) 规范化的 ERP (实线) 与规范化颗粒 LFP (虚线) 叠加。<a href="//cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/56447/56447fig7large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
ERP 信号是通过在头骨上放置一个有毛刺孔的蜘蛛电极来测量的。为了研究孔对 EEG 记录的影响, 另一只蜘蛛电极被放置在信息学侧筒皮层的完整颅骨上。注意, 以确保两个蜘蛛电极的阻抗是可比的大小, 通过调整的脑电图糊的数量使用。这里介绍了四只老鼠的数据 (它们不是上面所用的老鼠)。
图 8显示了从一只大鼠的两个电极进行的同时休眠状态 EEG 记录, 并显示在图 8A中的100s 数据, 而矩形框架 (二十年代) 中的数据则在图 8B中展开。这两种 EEG 信号在振幅相似的范围内有很大的共同变化。图 9显示了四只大鼠的 PSD, 顶行使用了频率轴上的线性刻度, 而在频率轴上使用对数刻度的底部行在低频范围内提供了扩展视图。从图 9中, PSD 中的主体之间似乎没有一致的偏差。在图 10中, 对五频段的平均 PSD 中的归一化差异进行了单样本 t 检验, 从而证实了这一点。这些频带上的归一化 PSD 差异与零 (p = 0.32、0.46、0.85、0.69 和0.97 分别) 没有显著的差别。
<img alt="Figure 8" class="xfigimg" src="/files/ftp_upload/56447/56447fig8.jpg"> 图 8: 双边脑电图记录.(A) 颅骨脑电图在休眠状态下, 在颅骨 (黑色) 上有一个毛刺孔, 同时在相对半球上同时进行脑电图记录 (灰色)。(B) (A) 中矩形框架内波形的扩展视图。<a href="//cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/56447/56447fig8large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 9" class="xfigimg" src="/files/ftp_upload/56447/56447fig9.jpg"> 图 9: 逆 (蓝色) 和信息学侧 (红色) 脑电图的功率谱密度 (PSD).每列显示的 PSD 为一只老鼠。顶部面板使用线性频率刻度, 而底部面板使用对数频率刻度, 使 PSD 在较低的频率范围内可视化。<a href="//cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/56447/56447fig9large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 10" class="xfigimg" src="/files/ftp_upload/56447/56447fig10.jpg"> 图 10: 组分析.信息学侧 PSD 与五频段的归一化差: 三角洲、θ、α、β和伽玛。每个条形图显示了在频带内的平均正常化差异, 标准错误显示为误差线。<a href="//cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/56447/56447fig10large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Concurrent Recording of Co-localized Electroencephalography and Local Field Potential in Rodent at JoVE.com

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

该协议描述了一种简单的方法, 并发记录的联合局部脑电图 (eeg) 和多层局部场电位在麻醉大鼠。在头骨上钻孔插入微电极的毛刺孔被证明能产生轻微的 EEG 信号失真。

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 ...

concurrent-recording-co-localized-electroencephalography-local-field

Universidad San Sebastian

Research

JoVE Journal

Neuroscience

12.2K 次观看.  University of Reading. 该协议描述了一种简单的方法, 并发记录的联合局部脑电图 (eeg) 和多层局部场电位在麻醉大鼠。在头骨上钻孔插入微电极的毛刺孔被证明能产生轻微的 EEG 信号失真。

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

Simultaneous fMRI and Electrophysiology in the Rodent Brain

To examine the neural basis of the blood oxygenation level dependent (BOLD) magnetic resonance imaging (MRI) signal, we have developed a rodent model in which functional MRI data and in vivo intracortical recording can be performed simultaneously. The combination of MRI and electrical recording is technically challenging because the electrodes used for recording distort the MRI images and the MRI acquisition induces noise in the electrical recording. To minimize the mutual interference of the two modalities, glass microelectrodes were used rather than metal and a noise removal algorithm was implemented for the electrophysiology data. In our studies, two microelectrodes were separately implanted in bilateral primary somatosensory cortices (SI) of the rat and fixed in place. One coronal slice covering the electrode tips was selected for functional MRI. Electrode shafts and fixation positions were not included in the image slice to avoid imaging artifacts. The removed scalp was replaced with toothpaste to reduce susceptibility mismatch and prevent Gibbs ringing artifacts in the images. The artifact structure induced in the electrical recordings by the rapidly-switching magnetic fields during image acquisition was characterized by averaging all cycles of scans for each run. The noise structure during imaging was then subtracted from original recordings. The denoised time courses were then used for further analysis in combination with the fMRI data. As an example, the simultaneous acquisition was used to determine the relationship between spontaneous fMRI BOLD signals and band-limited intracortical electrical activity. Simultaneous fMRI and electrophysiological recording in the rodent will provide a platform for many exciting applications in neuroscience in addition to elucidating the relationship between the fMRI BOLD signal and neuronal activity.

We have developed a method for simultaneous functional magnetic resonance imaging and electrophysiological recording in the rodent brain, providing a platform for the investigation of the relationship between neural activity and the blood oxygenation level dependent (BOLD) MRI signal.

同时在啮齿动物脑功能磁共振成像和电

To examine the neural basis of the blood oxygenation level dependent (BOLD) magnetic resonance imaging (MRI) signal, we have developed a rodent model in ...

科研

神经科学

Simultaneous Recordings of Cortical Local Field Potentials and Electrocorticograms in Response to Nociceptive Laser Stimuli from Freely Moving Rats

Electrocortical responses, elicited by laser heat pulses that selectively activate nociceptive free nerve endings, are widely used in many animal and human studies to investigate the cortical processing of nociceptive information. These laser-evoked brain potentials (LEPs) consist of several transient responses that are time-locked to the onset of laser stimuli. However, the functional properties of the LEP responses are still largely unknown, due to the lack of a sampling technique that can simultaneously record neural activities at the surface of the cortex (i.e., electrocorticogram [ECoG] and scalp electroencephalogram [scalp EEG]) and inside the brain (i.e., local field potential [LFP]). To address this issue, we present here an animal protocol using freely moving rats. This protocol is composed of three main procedures: (1) animal preparation and surgical procedures, (2) a simultaneous recording of ECoG and LFP in response to nociceptive laser stimuli, and (3) data analysis and feature extraction. Specifically, with the help of a 3D-printed protective shell, both ECoG and LFP electrodes implanted on the rat&#39;s skull were securely held together. During data collection, laser pulses were delivered on the rat&#39;s forepaws through gaps in the bottom of the chamber when the animal was in spontaneous stillness. Ongoing white noise was played to avoid the activation of the auditory system by the laser-generated ultrasounds. As a consequence, only nociceptive responses were selectively recorded. Using the standard analytical procedures (e.g., band-pass filtering, epoch extraction, and baseline correction) to extract stimulus-related brain responses, we obtained results showing that LEPs with a high signal-to-noise ratio were simultaneously recorded from ECoG and LFP electrodes. This methodology makes the simultaneous recording of ECoG and LFP activities possible, which provides a bridge of electrocortical signals at the mesoscopic and macroscopic levels, thereby facilitating the investigation of nociceptive information processing in the brain.

We developed a technique that simultaneously records both electrocorticography and local field potentials in response to nociceptive laser stimuli from freely moving rats. This technique helps establish a direct relationship of electrocortical signals at the mesoscopic and macroscopic levels, which facilitates the investigation of nociceptive information processing in the brain.

在自由运动大鼠的反应中, 皮质局部电位和电皮质图谱的同时记录

Electrocortical responses, elicited by laser heat pulses that selectively activate nociceptive free nerve endings, are widely used in many animal and ...

行为学

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

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls body functions and internal organ rhythms when animals are exposed to emotional challenges and changes in their living environments. In general experiments, signals from different organs, such as the brain and the heart, are recorded by independent recording systems that require multiple recording devices and different procedures for processing the data files. This study describes a new method that can simultaneously monitor electrical biosignals, including tens of local field potentials in multiple brain regions, electrocardiograms that represent the cardiac rhythm, electromyograms that represent awake/sleep-related muscle contraction, and breathing signals, in a freely moving rat. The recording configuration of this method is based on a conventional micro-drive array for cortical local field potential recordings in which tens of electrodes are accommodated, and the signals obtained from these electrodes are integrated into a single electrical board mounted on the animal's head. Here, this recording system was improved so that signals from the peripheral organs are also transferred to an electrical interface board. In a single surgery, electrodes are first separately implanted into the appropriate body parts and the target brain areas. The open ends of all of these electrodes are then soldered to individual channels of the electrical board above the animal's head so that all of the signals can be integrated into the single electrical board. Connecting this board to a recording device allows for the collection of all of the signals into a single device, which reduces experimental costs and simplifies data processing, because all data can be handled in the same data file. This technique will aid the understanding of the neurophysiological correlates of the associations between central and peripheral organs.

This study introduces a method for the simultaneous recording of local field potentials in the brain, electrocardiograms, electromyograms, and breathing signals of a freely moving rat. This technique, which reduces experimental costs and simplifies data analysis, will contribute to the understanding of the interactions between the brain and peripheral organs.

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 ...

Recording Spatially Restricted Oscillations in the Hippocampus of Behaving Mice

The local field potential (LFP) emerges from ion movements across neural membranes. Since the voltage recorded by LFP electrodes reflects the summed electrical field of a large volume of brain tissue, extracting information about local activity is challenging. Studying neuronal microcircuits, however, requires a reliable distinction between truly local events and volume-conducted signals originating in distant brain areas. Current source density (CSD) analysis offers a solution for this problem by providing information about current sinks and sources in the vicinity of the electrodes. In brain areas with laminar cytoarchitecture such as the hippocampus, one-dimensional CSD can be obtained by estimating the second spatial derivative of the LFP. Here, we describe a method to record multilaminar LFPs using linear silicon probes implanted into the dorsal hippocampus. CSD traces are computed along individual shanks of the probe. This protocol thus describes a procedure to resolve spatially restricted neuronal network oscillations in the hippocampus of freely moving mice.

This protocol describes the recording of local field potentials with multi-shank linear silicon probes. Conversion of the signals using current source density analysis allows the reconstruction of local electrical activity in the mouse hippocampus. With this technique, spatially restricted brain oscillations can be studied in freely moving mice.

行为小鼠海马空间受限振荡的记录

The local field potential (LFP) emerges from ion movements across neural membranes. Since the voltage recorded by LFP electrodes reflects the summed ...

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.

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 ...

小鼠运动皮层中的细胞外电生理记录

Multichannel Extracellular Recording in Freely Moving Mice

The protocol aims to uncover the properties of neuronal firing and network local field potentials (LFPs) in behaving mice carrying out specific tasks by correlating the electrophysiological signals with spontaneous and/or specific behavior. This technique represents a valuable tool in studying the neuronal network activity underlying these behaviors. The article provides a detailed and complete procedure for electrode implantation and consequent extracellular recording in free-moving conscious mice. The study includes a detailed method for implanting the microelectrode arrays, capturing the LFP and neuronal spiking signals in the motor cortex (MC) using a multichannel system, and the subsequent offline data analysis. The advantage of multichannel recording in conscious animals is that a greater number of spiking neurons and neuronal subtypes can be obtained and compared, which allows the evaluation of the relationship between a specific behavior and the associated electrophysiological signals. Notably, the multichannel extracellular recording technique and the data analysis procedure described in the present study can be applied to other brain areas when conducting experiments in behaving mice.

作者聚焦:用于研究自由移动小鼠神经元活动的多通道细胞外记录的进展 

The protocol describes the methodology of extracellular recording in the motor cortex (MC) to reveal extracellular electrophysiological properties in freely moving conscious mice, as well as the data analysis of local field potentials (LFPs) and spikes, which is useful for evaluating the network neural activity underlying behaviors of interest.

自由移动小鼠的多通道细胞外记录

The protocol aims to uncover the properties of neuronal firing and network local field potentials (LFPs) in behaving mice carrying out specific tasks by ...

小鼠大脑中的 LFP 记录用于认知研究和药物分析

Dual Extracellular Recordings in the Mouse Hippocampus and Prefrontal Cortex

The technique of recording local field potentials (LFPs) is an electrophysiological method used to measure the electrical activity of localized neuronal populations. It serves as a crucial tool in cognitive research, particularly in brain regions like the hippocampus and prefrontal cortex. Dual LFP recordings between these areas are of particular interest as they allow the exploration of interregional signal communication. However, methods for performing these recordings are rarely described, and most commercial recording devices are either expensive or lack adaptability to accommodate specific experimental designs. This study presents a comprehensive protocol for performing dual-electrode LFP recordings in the mouse hippocampus and the prefrontal cortex to investigate the effects of antipsychotic drugs and potassium channel modulators on LFP properties in these areas. The technique enables the measurement of LFP properties, including power spectra within each brain region and coherence between the two. Additionally, a low-cost, custom-designed recording device has been developed for these experiments. In summary, this protocol provides a means to record signals with high signal-to-noise ratios in different brain regions, facilitating the investigation of interregional information communication within the brain.

作者聚焦:使用双 LFP 记录方案研究药物对小鼠大脑信号的影响

This protocol outlines the use of a custom-designed recording device and electrodes to record local field potentials and investigate information flow in the mouse hippocampus and prefrontal cortex.

小鼠海马体和前额叶皮层中的双重细胞外记录

The technique of recording local field potentials (LFPs) is an electrophysiological method used to measure the electrical activity of localized neuronal ...

啮齿动物联合局部脑电图和局部电位的同步记录

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同时在啮齿动物脑功能磁共振成像和电

在自由运动大鼠的反应中, 皮质局部电位和电皮质图谱的同时记录

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

行为小鼠海马空间受限振荡的记录

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

自由移动小鼠的多通道细胞外记录

自由移动小鼠的多通道细胞外记录

自由移动小鼠的多通道细胞外记录

小鼠海马体和前额叶皮层中的双重细胞外记录

小鼠海马体和前额叶皮层中的双重细胞外记录