This work was supported by the Foundation for Anesthesia Education and Research Mentored Research Training Grant (ARM), by the National Institutes of Health grants GM107117 (MBK) and GM088156 (MBK), and by the Department of Anesthesiology and Critical Care at the University of Pennsylvania, Perelman School of Medicine.

整个调查进行的研究均符合健康指南全国学院协议，实验动物的护理和使用，并在宾夕法尼亚大学的机构动物护理和使用委员会批准。 1.头饰设计与施工<ol><li>由通过塑料砖推压销的插座部分移除有一对镊子的100位置的插座连接器的2×50针砖销的每一第八行。 注:朝下的引脚将是将为协议的其余部分引用的方向。 （请注意这特别是在2.6）。 </li><li>盖上的指甲油很轻的外衣引脚隔离，让指甲油完全干燥。 </li><li>从用丙酮和小布的销的尖端去除指甲油。 </li><li>用刀片或线切割p修剪多余的塑料的2×7的的liers。这将导致2×7被沿着销的长度绝缘并在针尖露出砖块。这些最终将成为经颅脑电图电极。两个2×7砖是必要的一个完整的慢性电极阵列（ 图1A）。 </li><li>削减EMG信号记录两个1 x 2针砖块。使用除去用镊子不需要的销和切割多余的塑料远以创建1×2砖的相同的过程。确保这些1×2的具有光滑面将它们从原来的100位置插座作为该距离将成为各头盔的标准销的距离，从而在单个适配器将所有头饰（ 图1A）工作。 </li><li>使用两部分环氧树脂的1×2针片附着到2 x 7针片（图1D）。 <ol><li>作为销的两个集都必须在同一方向，环氧树脂的1×2上的头盔的两半的横向侧与1×2和2×的平滑侧面7彼此接触。对齐1×2针孔和针与最靠后的2行上的2×7引脚。 注意:头巾的两半不环氧树脂粘合在一起。这允许对于习惯中更容易连接头状适配器的两半内灵活性，并在实验日（ 图1E）。 </li><li>让头盔半固化一夜。完成后，该头盔是左右对称的。每半年由一个2×7针砖与横向连接的2×1脚砖这是符合最后2行2×7针砖。 </li></ol></li><li>准备EMG信号记录电线。单链，31克全氟烷绝缘银线用于肌电信号记录（ 图1D）。然而，多股或者如果需要其他金属布线可以被取代。 <ol><li>要创建胸EMG电线取一块3.0厘米长的片全氟烷绝缘银线和REM的OVE1厘米从一端用刀片塑料的绝缘材料。环绕一对镊子的非绝缘导线的两倍。从镊子取出线，并删除在用刀片非环形端绝缘为25毫米。 </li><li>构建颈椎肌电图线，重复这个过程用铁丝的1.5厘米的段。都需要一个完整的头盔两节颈椎肌电图线和两个胸EMG线。 </li></ol></li><li>除去这两个头饰的最远前行中的横向销，其对应于3.3毫米前囟的和2.3毫米横向前囟的立体定位坐标，因为有这个位置下无脑作为由鼠标脑图谱20（ 图确定2A）。 </li><li>在两个半部分的头盔，切1×2砖的引脚与一对钢丝钳（从销的尖端3.0mm厚）的头盔的塑料基和焊接颈部肌电图线到前销和胸EMG的后验概率p在。 <ol><li>检查每个引脚的电隔离。由电压表的两根导线，以不同的引脚连接，而在连续性模式执行与数字万用表连续性测试。电气隔离引脚将不会产生与此万用表检测蜂鸣声;然而，电连接销会。覆盖焊点用指甲油和一次干燥，弯曲肌电电线，使得它们在平行于以最小的横向位移的前/后轴线。 </li></ol></li><li>修剪管脚的相对长度，使得它们匹配大脑的表面轮廓。 <ol><li>用鼠标脑图谱的助手，对于每个引脚脑表面记录腹侧距离前囟门坐标。20引脚，其腹侧距离前囟门是最大的将作为销修整指标。该引脚将不会被削减，而所有其他引脚将关于削减到这个最大腹针的距离（ 表1）。 注:引脚可磨下来的大小，但它必须精心为引脚和砂轮之间的摩擦可能会导致头盔的针弯曲来完成。如果针脚弯曲，用镊子进行纠正。向下研磨管脚长度的另一种方法是用一对线切割钳的修剪它们。 </li></ol></li><li>涵盖所有与使用银溶液笔银溶液针尖的，让干。这一步降低电极阻抗，以≤30kΩ的，这增加了信噪比，并同时消除了销微调引起的粗糙的边缘，因此，降低组织损伤的可能性和从手术加速恢复。已完成的头盔一半的权重约为0.5克</li></ol> 2.适配器建设通道映射<ol><li>切断的36位双排男纳米微型连接器的连接器的电线来的用刀片2或3厘米的统一的长度。每根电线，圣撕掉从端和锡为每个线暴露的金属绝缘2.5毫米。镀锡时有镀锡线的一条细链的每个纳米线适配器，因为这是隔离销至关重要确认。剪掉用铁丝剪切钳（ 图1C）剥离绝缘。 </li><li>创建使用匹配的公/母连接器连接到头盔的康涅狄格州大道头2×50切两2×7的和两个2×1的从2×50砖。通过断开男性引脚之一取下2×50砖不需要针，然后用镊子（ 图1B）推动同一块的完整下半时的连接器。 注意:这些引脚的一侧将作为适配器销插入到每个头盔，而另一半将被焊接到镀锡纳米连接器导线。确保有2×1,2×7感人的平板塑料边缘，以保证公/母连接器的正确匹配与步骤创建的头盔1。 </li><li>焊料上从纳米连接到所需的接地/基准销接地/参考线中的一个。地面和参考导线的RHD2132放大器芯片上捆绑在一起。使用单针是0.60毫米先于前囟门和前囟门为1.00mm横向既参考和地面。 （左头盔，第三最前行的内侧销;然而，任何其它管脚可以如优选分配， 图2）请注意，有可能通过去除0Ω电阻是关系地面放大器芯片上分开地和参考和参考，如果一起隔离两个期望。 </li><li>焊接镀锡纳米连接器的电线来的阳/阳连接器作为接地/基准销连接的同一侧。每个线映射到一个特定的信道，因此信道建立可以在这个时间内完成。为放大器的探头通道图图是在打开Ephys Wiki站点（https://open-ephys.atlassian.net/wiki/display/OEW/Home发现）。焊料，其信道是已知的相应的销，以实现所需的映射对应的导线。 </li><li>在与线切割钳纳米接头根部切断未使用的电线。 </li><li>使用电压表，以确保每个引脚与其他所有引脚的电隔离。一旦隔离确认，申请指甲油薄薄一层围绕每个焊点，以进一步隔离每个引脚。 </li><li>使用2部分环氧，加强配套纳米适配器对公对公砖双边2×7和2×1针。 注意:将有两部分，以匹配先前创建的头盔半的引脚排列，这种单一适配器。这是确保该适配器的每一半的中间部分不具有过量的环氧满溢阳/阳连接器的塑料边缘关键的，因为这将阻止头状的上下两部分被堵塞在同时进行。不要让任何环氧树脂流入针适配器POR底面适配器和灰，因为这也将妨碍正常连接。使用2头盔半为正确的连接器引脚排列的模具。 </li><li>环氧树脂适配器的两半和环氧纳米连接器，以增加其耐用性的基础。一定要覆盖所有焊点用环氧树脂。让适配器治愈过夜。 </li><li>可以用在开放式Ephys图形用户界面（GUI）阻抗测量进行适当的信道映射的确认。已完成的适配器的重量约130克（ 图 1F）。 </li></ol> 3.手术<ol><li> 准备一个无菌手术领域。 <ol><li>按要求戴无菌手套等个人防护装备。消毒在高压釜中的工具。消毒用1.0mM的二氧化氯溶液的立体定位框架。喷雾溶液到框架，并用无菌水漂洗之前等待5分钟。 </li><li>消毒植入式耳机套装IECE份，喷组件具有1.0毫米的二氧化氯溶液，并用无菌水漂洗之前等待5分钟。将现在无菌植入硬件到无菌培养皿。 </li></ol></li><li>得到一种手术前的重量为鼠标，然后麻醉小鼠中使用1.5-2.0％异氟烷在100％的氧气的200毫升感应腔室。使用的流速为大约500ml /分的腔室中。 </li><li>确认通过旋转感应室翻正反射损失。取下感应腔和地方变成立体定位框架的鼻锥鼠标不与耳棒完全固定鼠标的头部。继续监测供脚趾捏评价麻醉正确的深度，同时评估生命体征。 </li><li>维持核心体温在37℃用闭环温度控制器，诸如直肠探针和加热垫系统。修去毛皮前盖上眼科眼膏鼠标的眼睛使用弯剪刀或指甲刀的头顶。用消毒优碘头，并允许以优碘，然后再继续完全干燥。 </li><li>辖止痛药和抗生素液腹腔一起。对于25 G鼠标，0.5毫克的头孢唑啉，0.125毫克美洛昔康加入0.5ml生理盐水和2.5微克丁丙诺啡q 4-6小时PRN。 </li><li>注入沿着在头部中线250微升0.25％布比卡因皮下，和鼠标的两个颧弓注入100μl的0.25％布比卡因皮下。 </li><li>确保鼠标在立体框架，并暴露颅骨。 <ol><li>与立体耳酒吧的立体框架固定小鼠头部。确保鼠标在手术的麻醉平面通过确认不存在脚趾捏反射。创建具有沿着头骨中线11号一次性手术刀沿着1.5-2.0厘米的切口。切口将从眼睛之间开始，向后继续枕部。&lt;/ LI&gt; <li>通过微夹钳侧向皮肤暴露颅骨。从2.0％降低的异氟烷浓度维持麻醉的手术平面的浓度，但不降低到低于1.0％异氟烷在100％的氧。手术前镇痛减少维持麻醉的手术深度所需吸入麻醉剂的量，并且可以导致更快的恢复和改善的生存结果。 </li></ol></li><li> 水平头骨和钻颅骨钻孔。 <ol><li>识别前囟门和前囟门，成为坐标系的原点为零的立体定向坐标。成水平头骨在内侧/外侧轴，将附着到立体机械臂1.50毫米横向匀探针从囟两个方向，并确认背/腹深度小于0.05mm时探头接触的左，右头骨的侧面。 注:背/腹操纵AR的10微米分辨率M在一起使用的数字坐标显示简化整平。找平关于前囟的前/后轴线遵循相同的技术。用于接触前囟门和的Lamda在腹侧距离差也应小于0.05毫米。 <ol><li>调整颅骨直至匀是在两个方向上的完整，以使横向平面平行于地面。这允许所看到在小鼠脑图谱真实立体定向坐标。20 </li></ol></li><li>随着立体定向钻内的直径为0.5mm微型钻头，从3.30毫米前钻颅骨钻孔到4.50毫米后到前囟1.30 mm递增在横向1.00毫米对头骨的两半的中线。为电极的2.30毫米侧的列，从2.00毫米前钻头毛刺孔前囟点到4.50毫米后方前囟1.30 mm递增的中线的两侧（ 图2）。高准确度和精密度所需要这种博士解毒，阿莫西林操作由数字立体机械臂的10微米的分辨率简化。 注:为了使头盔的插针正确植入，鼠标的头骨必须在立体框架内牢固地到位。如果颅骨钻孔过程中移动时，头盔和颅骨钻孔的偏差可能会接踵而至。 </li></ol></li><li> 植入的头饰。 <ol><li>直镊子，准备EMG线隧道胸EMG电线。挖洞的皮肤和肌肉之间在2.5厘米的背面为左和右肌电导线两者。将胸椎和颈椎肌电图与直钳第一次创建腔，然后操纵脑电图砖弯钳使得销与之前钻出的毛刺孔对齐。 </li><li>轻压到头盔和扭动销插入头骨。销钉直径为0.46毫米。绝缘的指甲油，引脚将紧密贴合在钻毛刺浩LES。头状将是稳定一旦被适当地插入。调整EMG电线最终位置。重复同样的过程，以另一侧的头盔。 </li></ol></li><li> 安全使用牙科粘固粉到位头盔。 <ol><li>当两个头饰到位被设置时，混合比例为1:1的甲基丙烯酸甲酯与它的交联化合物。应用该混合物，使得它覆盖暴露颅骨，针电极的钉抛光部件，和EMG导线近端部分，但不覆盖头盔的阴插座。 </li><li>一定要皮毛得不到水泥。不允许水泥脊以形成鼠标就能抓住上。保证足够的时间使水泥干，然后从立体定位框架中移除鼠标。鼠标将不得不携带的总重量为从2半部的头盔的和固定的水泥是大约120克</li></ol></li><li> 放置动物在一个干净的恢复区。 <ol><li>维持核心体温用加热垫。监视鼠标，直到它重新获得所有姿势反射，标志着麻醉苏醒。个人住房建议长期复苏。 </li><li>最少3天手术后每日监测建议介入镇痛。开始习惯栓期前的复苏术后10-14天允许。 </li></ol></li></ol> 4. Habituate动物圈养<ol><li>将适配器连接到使用鼠标头枕（ 图1G-H）鼠标。抓住正在一旦鼠标被限制和适配器销慢慢插入两侧的植入头盔具有弯曲止血胶结到位头饰的相对的角落。 </li><li> 32通道放大器连接到适配器（ 图1H）。一定要调整适配器和放大器上都徽标的合作nsistent取向为适配器和防止信道映射误差放大器两者。放大器连接到RHD2000标准串行外设接口（SPI）电缆。该电缆将连接到信号记录的采集系统。 </li><li>鼠标放置在安装在室壁上的悬臂的腔室之内。附加SPI接口电缆连接到悬臂和调整在悬臂的张力抵消系留电缆的重量。鼠标可以自由移动和记录之前，习惯于一个小时，每天的一周。 </li><li>要断开鼠标，而在断开鼠标适配器使用平板不锈钢微锅铲助手只需拔下鼠标电缆和适配器。 </li></ol> 5.信号提取系统设置/信号记录<ol><li>请将构建适配器插入植入老鼠的头饰。一个探头放大器连接到适配器和附加标准的SPI接口电缆连接到功放和对采集板。有SPI的电缆连接到一个适当张紧悬臂使得在小鼠的头部的额外重量被最小化。 </li><li>将当地的法拉第笼，创建使用进行网或铝箔，周围的探头和地面局部法拉第笼。 </li><li>通过经由GUI中的模块中选择一个30 KS /秒的采样率和测量阻抗获得每个记录开始前电极的阻抗。小于或等于10千欧为单个引脚的阻抗值需要适当电极接触的确认。较高的阻抗值导致从电极拒绝的数据。 </li><li>用于记录，创建节奏的FPGA，带通滤波器，并在GUI LFP观看者的信号链。建议选择1.00 kS / s的，0.1-7,500赫兹的带宽，采样率和取消DSP。带通滤波器设置为0.1-250赫兹并显示由OP渠道厄宁的LFP观众。用抽签的方法250和400μV通道振幅选择最佳可视化的数据。 </li><li>开始使用GUI记录。为每个记录一个新的文件夹，并设置路径保存文件到该文件夹​​。要开始录制只需点击记录。从该连接器的所有32个信道是默认记录，但不需要的频道可以通过点击节奏FPGA模块的右侧记录开始之前被取消选择。 </li><li>导入数据到Matlab进行分析。有迹象表明，可用于分析，以帮助开源工具箱，众说纷纭。 </li></ol>

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The authors have nothing to disclose.

低成本结构和必要的，以便正确地实现在小鼠26信道，高密度脑电图蒙太奇外科步骤进行说明。正确的硬膜外电极接触是在该系统中获取优质脑电信号的关键。协议地址中的两个步骤此问题:脚微调，以配合大脑的轮廓，以及头盔植入前亚克力加固。不要切针，在施工阶段太短，这一点很重要。当植入头饰，当务之急是最终的丙烯酸加固前检查管脚布局。以确认适当的电极接触的一个方法是通过阻抗测试。从表面上看，5-10千欧的阻抗，建议适当硬膜外安置。26 &#160; 阻抗测量证明头饰&#39;的耐用性，如电极的阻抗值是这个5-10千欧范围为植入后至少4个月内保持稳定。另一个关键步骤涉及对准EMG引脚与2×7脑电图砖的两个最靠后的行。这是为适配器连接关键，因为未对准肌电图和脑电图销将导致无法连接适配器或弯曲适配器销。 本获取系统的主要优点是易于修改电极阵列的形状，以优化不同的实验需要的。被最佳地适合于特定的实验定制电极的安排可以容易地创建。定制为特定的实验可能与插管用于定向药物递送用于组合的药理学，脑电图，和行为研究结合脑电图27头饰，适配器和外科手术容易针对以下在协议上述方法时宽一些研究。本次收购系统的第二大优势是其成本低。目前，本次收购系统能多达4个单独的电缆记录128个输入通道，从4只小鼠或者如果需要的话，只用更高密度的网格，允许同时进行的录像。这种扩大将只需要额外的电缆和适配器。 这种方法的高密度脑电图采集地址的小鼠其它高密度脑电图采集方法的缺点。在这项工作中所描述的系统被轻易地用简单的材料构造并使用开放源码的硬件和软件，廉价且稳定，允许在相同的动物重复测量在数月，在实验期间允许自由运动，并且不需要小鼠是麻醉记录。本系统的限制是，它仅被验证迄今为止在该称量20g以上，并且是老年人超过12周的小鼠。更小或更年轻的小鼠可能与头盔植入困难。这种方法的第二个局限性是不能精确控制耳机套装后，电极深度IECE制造。然而，这种相同的限制适用于传统的螺钉EEG电极，因为没有办法精确地知道验预螺纹深度相对于该皮层的表面上。故障排除这种方法通常为了得到无噪声信号栓时涉及正确地从小鼠屏蔽干扰信号。 高密度脑电图阵列是对于那些在现代脑电图解读新脑电图正常复杂的数据分析时空必不可少。同时示出一个视觉诱发电位的空间分布，使用该系统获得的数据可利用电源的成像技术和神经元的连接的措施来进行分析。在接触区域A的60％到70％的减少，这些电极销之间相对于传统的螺钉接触允许更精确的信号定位， 如图4。用人转基因小鼠中的高密度的分析技术，以下药理学ogical干预，或与内在的病理动物，如癫痫病可以帮助辨别生成特定的皮质振荡的机制，本地化的ERP和EP唱片的来源，揭示大型网络性能。通过更好地并联人类系统，这种方法将提高人类神经生理学和神经病理学的小动物模型，提供更容易在啮齿动物模型中人类作出科学和临床相关性的发现翻译。

神经元的活动可以与细胞外各种级别从微观（单个动作电位）粒度介观（局部场电位）的宏观（脑电图）进行记录。这些脑电波痕迹分析经典在频域表征行为，神经生理学，或电生理状态。这可以用一个单一的生物电势，4-但疏密度脑电图记录做不能解决神经元活动的空间分量。现代脑电图分析依赖于多个电极，以产生皮层活动的时空分布的详细地图，以便与具体的心理状况和生理过程该活动相关联。分析的需要高密度脑电图蒙太奇是更常用的类别的5-7两源电器成像和神经网络的连通性的措施。8-11 12-15由于脑电图具有很高的时间分辨率，脑电图研究允许的ERP和EP唱片的实时评估，以及精确的时间事后分析。3,11 ，12 关联认知状态和功能与上看到的脑电图振荡的相互作用是神经网络连通性的各项措施的最终目标。大量研究显示，同步和与觉醒，注意力和行动的具体状态相关的不同脑区之间振荡的锁相。6,13,14,16-19 </sup&gt;脑区之间的示范这种信号协会要求的高密度阵列，允许网络连接的评估。 EEG信号的源定位和网络分析源自于人类的研究，但调查这些信号的神经元基础必然涉及动物模型，因为它们需要是在人类中，否则不可能侵入技术。为了复制在啮齿动物模型中这些分析，需要用于在啮齿动物的脑中捕获高密度的EEG信号的方法。而其它基团已经构建高密度微阵列用于在小鼠中使用，这样的方法是有限的研究人员无法获得纳米加工设施，不适合反复实验足够健壮在延长的时间周期，或者被限制在麻醉使用小鼠1-3,7构建慢性高密度的低成本替代方案，经颅电极ARRAy为这里演示。 这里描述的信号采集的方法并不限定于脑电图，但包括肌电图（EMG）信号。肌电信号采集可以定义的行为状态的补充办法，是睡眠研究特别有用。这种方法提供了昂贵，超高密度颅内网格之间的中间，并且可能与传统的螺旋电极是不够的高级分析办法的有限导数。该头盔的设计是很容易构造和负担得起的高通量的研究。在啮齿动物模型中各类遗传或药理操纵技术可以帮助发现皮质振荡代，从真基因型差异的行为的分歧，事件相关电位和EP唱片的源定位，以及大规模网络通信机制结合使用该采集系统。

<table><tbody><tr><td>32 Channel RHD2132 amplifier headstage</td><td>Intan Technologies</td><td>C3314</td><td></td></tr><tr><td>Aquistion Board</td><td>Open Ephys</td><td>v2.2</td><td></td></tr><tr><td>100 Position Receptable Connector</td><td>Digi-Key</td><td>ED85100-ND</td><td>Headpiece</td></tr><tr><td>Acetone (1 L)</td><td>Sigma Aldrich</td><td>179973-1L</td><td></td></tr><tr><td>Razor Blade (100 pack)</td><td>McMaster Carr</td><td>3962A4</td><td></td></tr><tr><td>Wire-Cutting Pliers</td><td>MSC Industrial</td><td>321786</td><td></td></tr><tr><td>2-Part Epoxy</td><td>McMaster Carr</td><td>7605A18</td><td></td></tr><tr><td>PFA Coated Silver Wire (25 ft)</td><td>A-M Systems</td><td>787000</td><td>EMG Wire</td></tr><tr><td>CircuitWriter Pen</td><td>MCM Electronics</td><td>200-175</td><td>Silver Applicator for Electrode Tips</td></tr><tr><td>36 Position Dual Row Male Nano-Miniature Connector</td><td>Omnetics Connector Corporation</td><td>A79028-001</td><td>Headpiece to Amplifier Adapter</td></tr><tr><td>Conn Strip Header 2 x 50</td><td>Digi-Key</td><td>ED83100-ND</td><td>Headpiece to Amplifier Adapter</td></tr><tr><td>Clidox Base and Acitvator</td><td>Pharmacal</td><td>95120F &amp;amp; 96120F</td><td>Sterilant</td></tr><tr><td>Isoflurane</td><td>Priamal Enterprises Ltd</td><td>66794-019-10</td><td></td></tr><tr><td>Oxygen</td><td>Airgas</td><td>OX USP300</td><td></td></tr><tr><td>Closed Loop Temperature Controller</td><td>CWE Inc.&amp;nbsp;</td><td>08-130000</td><td></td></tr><tr><td>Curved Scissors</td><td>FST</td><td>14085-09</td><td></td></tr><tr><td>0.25% Bupivicaine Hydrochloride</td><td>Hospira</td><td>0409-1159-02</td><td>Local Anesthetic</td></tr><tr><td>Meloxicam 5mg/ml</td><td>Henry Schein</td><td>6451602845</td><td>Pain/Inflammation Relief</td></tr><tr><td>0.9% Sodium Chloride</td><td>Hospira</td><td>0409-4888-20</td><td>Fluids</td></tr><tr><td>Cefazolin</td><td>Hospira</td><td>0409-0806-01</td><td>Antibacterial</td></tr><tr><td>No.11 Disposable Scapel (20 pk)</td><td>Feather</td><td>2975#11</td><td></td></tr><tr><td>Micro Serrefines</td><td>FST</td><td>18052-3</td><td></td></tr><tr><td>Cotton Swabs (1,000 pk)</td><td>MSC Industrial</td><td>8749574</td><td></td></tr><tr><td>0.5 mm Micro Drill Bit</td><td>FST</td><td>19007-05</td><td></td></tr><tr><td>Stereotaxic Drill</td><td>Kopf</td><td>Model 1471</td><td></td></tr><tr><td>Curved Forceps</td><td>Roboz</td><td>RS-5136</td><td></td></tr><tr><td>Methyl Methacrylate</td><td>A-M Systems</td><td>525000</td><td>Cement for headpiece</td></tr><tr><td>Methyl Methacrylate Crosslinking Compound</td><td>A-M Systems</td><td>526000</td><td></td></tr><tr><td>Curved Hemostats</td><td>FST</td><td>13003-10</td><td>Aide in Adapter Connection</td></tr><tr><td>RHD2000 standard SPI interface cable (3ft)</td><td>Intan Technologies</td><td>C3203</td><td></td></tr><tr><td>Cantilever Arm</td><td>Instech</td><td>MCLA</td><td></td></tr><tr><td>Micro Spatula (12 pk)</td><td>Fischer Scientific</td><td>S50822</td><td></td></tr><tr><td>Digital Soldering Station</td><td>MCM Electronics</td><td>21-10115</td><td></td></tr><tr><td>Rosin Core Solder 60/40 Tin/Lead</td><td>MCM Electronics</td><td>21-1045</td><td></td></tr><tr><td>Color Craze Nail Polish with Hardeners (Nitrocellulose based)</td><td>L.A. Colors</td><td>CNP508</td><td></td></tr><tr><td>Small Animal Stereotaxic Instrument with Digital Display Console</td><td>Kopf</td><td>Model 940</td><td></td></tr></tbody></table>

high-density electroencephalographic acquisition in a rodent model using low-cost and open-source resources

需要高空间分辨率，包括电气源成像和网络连通性的措施，先进的脑电图分析技术，适用于不断扩大的各种神经科学的问题。在啮齿动物模型执行这些类型的分析需要更高的电极密度比传统的螺旋电极可以完成。而存在啮齿类更高密度脑电图蒙太奇，它们的有限可用性大多数研究人员，不适合反复实验足够健壮在延长的时间周期，或者被限制在麻醉的啮齿动物使用1-3拟议的低成本为构建一个持久的，高支，经颅电极阵列，包括双边植入头饰的方法进行了研究作为一种手段来执行在小鼠或大鼠先进的脑电图分析。 程序头盔制造和手术植入ñ以具完整产生高的信噪比，低阻抗脑电图和肌电图的信号出现。虽然该方法在大鼠和小鼠有用，这份手稿的重点是更具挑战性实施小老鼠头骨。自由移动小鼠经由公共适配器记录期间仅拴电缆。该电极系统，该系统包括26个脑电图频道和4肌电通道之一的版本进行说明。

记录在具有高密度的脑电图头盔植入自由移动小鼠样本数据示于图3。个别脑电图波形对应于图2所示的信道映射方法。颈椎和胸椎肌电图的实例也显示在图3中。需要注意的是胸EMG记录还包含嵌入式电活动起源于老鼠的心脏时，两个计算胸EMG线之间（T）的差分信号变得显而易见。与此记录，也可以通过测量心电图的QRS峰值之间的时间来计算小鼠的心脏速率23-24同样，有可能通过计算QRS峰值的相位变化的胸腔扩展来测量小鼠的呼吸频率和每次呼吸的合同。25因此，这种设置允许收购Ø˚F鼠多导睡眠监测。此外，设置使视觉诱发电位（ 图4）的皮质映射。当光的10毫秒脉冲只传递到鼠标的左眼，经典的反应被记录在对侧（但不同侧）初级视觉皮层被随后在对侧次级视觉皮层一个延迟的响应。嵌入图4中的动画显示的时间与在对侧V1和V2的活性的曲线图沿整个皮质表面不同的电位。 <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td>美联社</td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td> 3.3 </td><td></td><td></td><td> 0 </td><td> 0 </td><td></td><td></td></tr><tr><td> 2 </td><td></td><td> 0.4 </td><td> 0.6 </td><td> 0.6 </td><td> 0.4 </td><td></tD&gt; </tr><tr><td> 0.7 </td><td></td><td> 0.6 </td><td> 0.9 </td><td> 0.9 </td><td> 0.6 </td><td></td></tr><tr><td> -0.6 </td><td></td><td> 0.9 </td><td> 1 </td><td> 1 </td><td> 0.9 </td><td></td></tr><tr><td> -1.9 </td><td></td><td> 1 </td><td> 1.1 </td><td> 1.1 </td><td> 1 </td><td></td></tr><tr><td> -3.2 </td><td> 3 </td><td> 1 </td><td> 1 </td><td> 1 </td><td> 1 </td><td> 3 </td></tr><tr><td> -4.5 </td><td> 3 </td><td> 0.7 </td><td> 0.7 </td><td> 0.7 </td><td> 0.7 </td><td> 3 </td></tr><tr><td> ML </td><td></td><td> -2.3 </td><td> -1 </td><td> 1 </td><td> 2.3 </td><td></td></tr></tbody></table> 表1:引脚长度修剪此图显示了需要微调的长度，单位为毫米，每针的头饰。引脚长度修剪从老鼠大脑图谱收购。房颤之三修整引脚，头盔大脑的表面轮廓相匹配。如用于记录EMG信号的导线被焊接在销存根20肌电销被完全切断。 <img alt="图1" src="/files/ftp_upload/54908/54908fig1.jpg" /> 图 1: 头饰组件，中级建设步骤，以及用于记录正常连接此图显示了用于创建头饰的原料。用100针插座连接器开始，创建更小的2×7和2×1分量。另外，在2×1分量，的2×50原边是完整的，其允许一致头盔结构，并允许一个适配器连接到许多植入小鼠中。 图1B和1C介绍必要创建适配器原材料从头盔到放大器。1B呈现的头盔结束相似地被砍倒的适配器连接到头盔。需要注意的是2×1再次有从原始成分的原边，从而确保在适配器和头盔之间的正确连接。 图1C示出了一个连接到放大器的适配器的端部。 图1D示出了用环氧树脂2×7和2与制备的肌电电线为信号记录沿着×1的部件。 图1E说明了一个完整的头盔。 图1F显示完成适配器。 图1G示出了耳机和适配器之间的正确连接。最后， 图1H显示了植入式鼠标的连接适配器和放大器。该放大器芯片连接到运行到采集板（未显示）的接口电缆。 <a href="http://ecsource.jove.com/files/ftp_upload/54908/54908fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <p类="jove_content"FO: - together.within页保留="1"&gt; <img alt="图2" src="/files/ftp_upload/54908/54908fig2.jpg" /> 图 2: 电极蒙太奇和完全构造头饰该图显示相对于鼠脑电极放置。电极位置基于从囟立体坐标。每个电极坐标可以在协议步骤4.8中找到。电极颜色对应于每个电极的底层的大脑区域。白色=额叶联合皮层（FRA），橙色=初级运动皮层（M1），粉红=次级运动皮层（M2），深绿色=初级躯体感觉皮层，前肢区域（S1FL），绿色=初级躯体感觉皮层，dysgranular区（S1DZ ），浅绿色=初级躯体感觉皮层，每桶场（S1BF），黄色=内侧顶叶联合皮质（MPTA），暗蓝色=初级视觉皮层（V1），浅蓝色=次级视觉皮层，mediomedial区（V2MM），黑色= retrosplenial DYsgranular皮层（RSD）20通用参考/地显示为好。该参考方案最大限度地减少了原始信号中的呼吸假象。与每个电极相关联的号码提供整个阵列的频道映射。图片来自艾伦鼠脑图谱。21,22 图2B修改显示了一个完全构造头盔相对于一毛钱规模。 <a href="http://ecsource.jove.com/files/ftp_upload/54908/54908fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/54908/54908fig3.jpg" /> 图3: 示例EEG和EMG从电极蒙太奇跟踪电极的波形对应于图1A中所示的频道映射。颈椎肌电图（C）提供了确定颈部肌张力（+）的能力。 EMG信号也含有心脏QRS电脉冲（*）。 200μV微量幅度和1秒，持续跟踪比例尺显示， <a href="http://ecsource.jove.com/files/ftp_upload/54908/54908fig3large.jpg" target="_blank">请点击这里查看该图的放大版本。</a> <img alt="图4" src="/files/ftp_upload/54908/54908fig4.jpg" /> 图4:视觉诱发电位的空间分布只给予左眼单方面光芒一闪的诱发电位以下应用的空间分布。上图描述了高密度的脑电图蒙太奇与代表电极每一个圆圈。在颜色随时间的变化对应于电压随时间的每个相应电极的变化。在时间= 0毫秒，10毫秒光脉冲被输送，并在中间图表示。下图说明意味着对侧V1和V2脑电图电极（N = 108 EP试验）诱发电位痕迹。光普勒在本身0毫秒出现。需要注意的是相应的诱发电位反应对侧V1（黑色线）观察，其次是潜伏期较长诱发对侧V2（红色曲线）的潜在反应。 <a href="https://www.jove.com/files/ftp_upload/54908/Figure_4.mov" target="_blank">（右键点击下载）。</a>

Watch this Scientific Journal Video about High-density Electroencephalographic Acquisition in a Rodent Model Using Low-cost and Open-source Resources at JoVE.com

High-density Electroencephalographic Acquisition in a Rodent Model Using Low-cost and Open-source Resources

Instructions for the low-cost construction and surgical implantation of a chronic transcranial high-density electroencephalographic montage into mice are provided. Signal recording, extraction, and processing techniques are also described.

高密度的脑电图采集在啮齿动物模型使用低成本的开源资源

Advanced electroencephalographic analysis techniques requiring high spatial resolution, including electrical source imaging and measures of network ...

high-density-electroencephalographic-acquisition-rodent-model-using

Research

JoVE Journal

Neuroscience

15.8K 次观看.  University of Pennsylvania. Instructions for the low-cost construction and surgical implantation of a chronic transcranial high-density electroencephalographic montage into mice are provided. Signal recording, extraction, and processing techniques are also described.

视频: High-density Electroencephalographic Acquisition in a Rodent Model Using Low-cost and Open-source Resources

High-density EEG Recordings of the Freely Moving Mice using Polyimide-based Microelectrode

Electroencephalogram (EEG) indicates the averaged electrical activity of the neuronal populations on a large-scale level. It is widely utilized as a noninvasive brain monitoring tool in cognitive neuroscience as well as a diagnostic tool for epilepsy and sleep disorders in neurology. However, the underlying mechanism of EEG rhythm generation is still under the veil. Recently introduced polyimide-based microelectrode (PBM-array) for high resolution mouse EEG1 is one of the trials to answer the neurophysiological questions on EEG signals based on a rich genetic resource that the mouse model contains for the analysis of complex EEG generation process. This application of nanofabricated PBM-array to mouse skull is an efficient tool for collecting large-scale brain activity of transgenic mice and accommodates to identify the neural correlates to certain EEG rhythms in conjunction with behavior. However its ultra-thin thickness and bifurcated structure cause a trouble in handling and implantation of PBM-array. In the presented video, the preparation and surgery steps for the implantation of PBM-array on a mouse skull are described step by step. Handling and surgery tips to help researchers succeed in implantation are also provided.

In this article, we described the surgery procedure and handling tips for implantation of ultra-thin polyimide-based microelectrode array (PBM-array) on the mouse skull for acquisition of high-density encephalography (EEG) in a mouse model.

高密度的自由移动，使用基于聚酰亚胺的微电极的小鼠的脑电图记录

Electroencephalogram (EEG) indicates the averaged electrical activity of the neuronal populations on a large-scale level. It is widely utilized as a ...

科研

神经科学

Inducing Post-Traumatic Epilepsy in a Mouse Model of Repetitive Diffuse Traumatic Brain Injury

Traumatic brain injury (TBI) is a leading cause of acquired epilepsy. TBI can result in a focal or diffuse brain injury. Focal injury is a result of direct mechanical forces, sometimes penetrating through the cranium, creating a direct lesion in the brain tissue. These are visible during brain imaging as areas with contusion, laceration, and hemorrhage. Focal lesions induce neuronal death and glial scar formation and are present in 20%&minus;25% of all people who incur a TBI. However, in the majority of TBI cases, injury is caused by acceleration-deceleration forces and subsequent tissue shearing, resulting in nonfocal, diffuse damage. A subpopulation of TBI patients continues to develop post-traumatic epilepsy (PTE) after a latency period of months or years. Currently, it is impossible to predict which patients will develop PTE, and seizures in PTE patients are challenging to control, necessitating further research. Until recently, the field was limited to only two animal/rodent models with validated spontaneous post-traumatic seizures, both presenting with large focal lesions with massive tissue loss in the cortex and sometimes subcortical structures. In contrast to these approaches, it was determined that diffuse TBI induced using a modified weight drop model is sufficient to initiate development of spontaneous convulsive and non-convulsive seizures, even in the absence of focal lesions or tissue loss. Similar to human patients with acquired post-traumatic epilepsy, this model presents with a latency period after injury before seizure onset. In this protocol, the community will be provided with a new model of post-traumatic epilepsy, detailing how to induce diffuse non-lesional TBI followed by continuous long-term video-electroencephalographic animal monitoring over the course of several months. This protocol will detail animal handling, the weight drop procedure, the electrode placement for two acquisition systems, and the frequent challenges encountered during each of the steps of surgery, postoperative monitoring, and data acquisition.

This systematic protocol describes a new animal model of post-traumatic epilepsy after repetitive mild traumatic brain injury. The first part details steps for traumatic brain injury induction using a modified weight drop model. The second part provides instructions on the surgical approach for single- and multi-channel electroencephalographic data acquisition systems.

在重复性扩散创伤性脑损伤小鼠模型中诱导创伤后癫痫

Traumatic brain injury (TBI) is a leading cause of acquired epilepsy. TBI can result in a focal or diffuse brain injury. Focal injury is a result of ...

行为学

Long-term Continuous EEG Monitoring in Small Rodent Models of Human Disease Using the Epoch Wireless Transmitter System

Many progressive neurologic diseases in humans, such as epilepsy, require pre-clinical animal models that slowly develop the disease in order to test interventions at various stages of the disease process. These animal models are particularly difficult to implement in immature rodents, a classic model organism for laboratory study of these disorders. Recording continuous EEG in young animal models of seizures and other neurological disorders presents a technical challenge due to the small physical size of young rodents and their dependence on the dam prior to weaning. Therefore, there is not only a clear need for improving pre-clinical research that will better identify those therapies suitable for translation to the clinic but also a need for new devices capable of recording continuous EEG in immature rodents. Here, we describe the technology behind and demonstrate the use of a novel miniature telemetry system, specifically engineered for use in immature rats or mice, which is also effective for use in adult animals.

Here we demonstrate the use of a wireless enabling technology for electroencephalogram (EEG) in neonatal rodent models of human disease. With telemetry, there are no encumbering connections, thus allowing natural behaviors.

在人类疾病中使用大纪元无线传输系统的小型啮齿动物模型的长期连续监测脑电图

Many progressive neurologic diseases in humans, such as epilepsy, require pre-clinical animal models that slowly develop the disease in order to test ...

Brain Source Imaging in Preclinical Rat Models of Focal Epilepsy using High-Resolution EEG Recordings

Electroencephalogram (EEG) has been traditionally used to determine which brain regions are the most likely candidates for resection in patients with focal epilepsy. This methodology relies on the assumption that seizures originate from the same regions of the brain from which interictal epileptiform discharges (IEDs) emerge. Preclinical models are very useful to find correlates between IED locations and the actual regions underlying seizure initiation in focal epilepsy. Rats have been commonly used in preclinical studies of epilepsy1; hence, there exist a large variety of models for focal epilepsy in this particular species. However, it is challenging to record multichannel EEG and to perform brain source imaging in such a small animal. To overcome this issue, we combine a patented-technology to obtain 32-channel EEG recordings from rodents2 and an MRI probabilistic atlas for brain anatomical structures in Wistar rats to perform brain source imaging. In this video, we introduce the procedures to acquire multichannel EEG from Wistar rats with focal cortical dysplasia, and describe the steps both to define the volume conductor model from the MRI atlas and to uniquely determine the IEDs. Finally, we validate the whole methodology by obtaining brain source images of IEDs and compare them with those obtained at different time frames during the seizure onset.

This video introduces the preparation, recording, and source analysis procedures of high-resolution EEG on sedated rats with a particular preclinical model of focal epilepsy under noninvasive conditions.

采用高分辨率脑电图记录脑源成像局灶性癫痫的临床前模型大鼠

Electroencephalogram (EEG) has been traditionally used to determine which brain regions are the most likely candidates for resection in patients with ...

医学

Polygraphic Recording Procedure for Measuring Sleep in Mice

Recording of the epidural electroencephalogram (EEG) and electromyogram (EMG) in small animals, like mice and rats, has been pivotal to study the homeodynamics and circuitry of sleep-wake regulation. In many laboratories, a cable-based sleep recording system is used to monitor the EEG and EMG in freely behaving mice in combination with computer software for automatic scoring of the vigilance states on the basis of power spectrum analysis of EEG data. A description of this system is detailed herein. Steel screws are implanted over the frontal cortical area and the parietal area of 1 hemisphere for monitoring EEG signals. In addition, EMG activity is monitored by the bilateral placement of wires in both neck muscles. Non-rapid eye movement (Non-REM; NREM) sleep is characterized by large, slow brain waves with delta activity below 4 Hz in the EEG, whereas a shift from low-frequency delta activity to a rapid low-voltage EEG in the theta range between 6 and 10 Hz can be observed at the transition from NREM to REM sleep. By contrast, wakefulness is identified by low- to moderate-voltage brain waves in the EEG trace and significant EMG activity.

The recording of electroencephalogram (EEG) and electromyogram (EMG) in freely behaving mice is a critical step to correlate behavior and physiology with sleep and wakefulness. The experimental protocol described herein provides a cable-based system for acquiring EEG and EMG recordings in mice.

印刷业记录程序在小鼠体内测量睡眠

Recording of the epidural electroencephalogram (EEG) and electromyogram (EMG) in small animals, like mice and rats, has been pivotal to study the ...

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

Although electroencephalography (EEG) is widely used as a non-invasive technique for recording neural activities of the brain, our understanding of the neurogenesis of EEG is still very limited. Local field potentials (LFPs) recorded via a multi-laminar microelectrode can provide a more detailed account of simultaneous neural activity across different cortical layers in the neocortex, but the technique is invasive. Combining EEG and LFP measurements in a pre-clinical model can greatly enhance understanding of the neural mechanisms involved in the generation of EEG signals, and facilitate the derivation of a more realistic and biologically accurate mathematical model of EEG. A simple procedure for acquiring concurrent and co-localized EEG and multi-laminar LFP signals in the anesthetized rodent is presented here. We also investigated whether EEG signals were significantly affected by a burr hole drilled in the skull for the insertion of a microelectrode. Our results suggest that the burr hole has a negligible impact on EEG recordings.

This protocol describes a simple method for concurrent recording of co-localized electroencephalography (EEG) and multi-laminar local field potential in an anesthetized rat. A burr hole drilled in the skull for the insertion of a microelectrode is shown to produce negligible distortion of the EEG signal.

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 ...

用低成本的 EEG 和毫米级线圈刺激小鼠大脑

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

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.

作者聚焦: 低成本脑电图记录系统结合毫米级线圈经颅刺激小鼠体内大脑  

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.

低成本脑电图记录系统结合毫米级线圈，在体内经颅刺激小鼠大脑

A low-cost electroencephalographic (EEG) recording system is proposed here to drive transcranial magnetic stimulation (TMS) of the mouse brain in vivo, ...

<meta charset="utf8"></head><body>用于大鼠多区域电生理监测的开源植入物

The TD Drive: A Parametric, Open-Source Implant for Multi&#45;Area Electrophysiological Recordings in Behaving and Sleeping Rats

Intricate interactions between multiple brain areas underlie most functions attributed to the brain. The process of learning, as well as the formation and consolidation of memories, are two examples that rely heavily on functional connectivity across the brain. In addition, investigating hemispheric similarities and/or differences goes hand in hand with these multi-area interactions. Electrophysiological studies trying to further elucidate these complex processes thus depend on recording brain activity at multiple locations simultaneously and often in a bilateral fashion. Presented here is a 3D-printable implant for rats, named TD Drive, capable of symmetric, bilateral wire electrode recordings, currently in up to ten distributed brain areas simultaneously. The open-source design was created employing parametric design principles, allowing prospective users to easily adapt the drive design to their needs by simply adjusting high-level parameters, such as anterior-posterior and mediolateral coordinates of the recording electrode locations. The implant design was validated in n = 20 Lister Hooded rats that performed different tasks. The implant was compatible with tethered sleep recordings and open field recordings (Object Exploration) as well as wireless recording in a large maze using two different commercial recording systems and headstages. Thus, presented here is the adaptable design and assembly of a new electrophysiological implant, facilitating fast preparation and implantation.

<meta charset="utf8"></head><body>作者聚焦：通过神经植入物破译记忆和学习以进行多区域大脑研究

Here, we present a unique, 3D-printable implant for rats, named TD Drive, capable of symmetric, bilateral wire electrode recordings, currently in up to ten distributed brain areas simultaneously.

TD Drive：一种参数化开源植入物，用于行为和睡眠大鼠的多区域电生理记录

Intricate interactions between multiple brain areas underlie most functions attributed to the brain. The process of learning, as well as the formation and ...

高密度的脑电图采集在啮齿动物模型使用低成本的开源资源

摘要

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TD Drive：一种参数化开源植入物，用于行为和睡眠大鼠的多区域电生理记录