这项工作是由美国国防部高级研究计划局（DARPA）MTO杰克朱迪博士和Doug Weber博士通过空间和海战系统中心，太平洋格兰特/合同No.N66001-12-C-4173的主持下举办。我们要感谢托马斯·埃格斯他在制造过程中的帮助，罗纳德Triolo，马修·席费尔，李费舍尔和Freeburg的最大值为他们在复合神经袖口设计的发展做出贡献。

1.电极组件准备<ol><li>收集需要精密切割四级式电极组件（使用激光切割，请参考材料清单）的制造过程之前。这些组件（ 图1）: 联系方式阵列框架:这个框架是做出来的125微米厚的聚醚醚酮（PEEK）板材。它覆盖袖带的整个宽度并保持中间触点，具有蛇形边缘（ 图1B）。中间接触被包裹在所述导向通道;因此触点的暴露宽度由信道的宽度的限制和间隔由信道之间的间隔来确定。 中间接点条:中间触点由缠绕触头阵列框架（ 图1B），这些条带形成。切带出铂/ 10％铱片到引导通道的宽度和添加额外的长度，以允许他们到bË周围的框架完全折叠。在0°角条的长轴点焊联系人的领先地位。 参考联系人:需要四个引用。这些接触的长尺寸比压脉袋的宽度稍短，充分包含它们的压脉袋的内部。点焊每个参考接触到铅在90度角与该联系人的主轴线。 PEEK间隔物:间隔被用来在电极上创建较薄区域，以允许弯曲和关闭（ 图1C）。所有的间隔物从PEEK（也可使用其它材料）制成并切成电极的长度。中间空间的宽度是相等的电极的高度。 </li></ol> 2.联系人阵列的制备<ol><li>清洗在步骤1通过超声处理在乙醇中在40千赫和室温，然后在相同的超声参数在去离子蒸馏水2分钟制成2分钟的组分。让干燥。 </li><li>目视检查联系方式对于像激光切割残留或表面变形的缺陷。 </li><li>定位接触逐个与焊接点朝上，在显微镜下。保持用镊子在大约长度起点的1/3从自由端接触。提升导致一个45度角，同时保持接触，使第一弯。 </li><li>放置与焊接朝上阵列框架下方的预先弯曲的接触。按住框架下用镊子和提升领先优势扩大到一个45度角进行第二次弯曲。而继续保持架向下，抓住与镊子和弯曲的接触在一个180度的角度（朝向框的中间线折叠）的自由端。 </li><li>伸直和拉向运营商接触，然后在180度的角度（折叠中间线）弯曲。点焊点现在应该在两个弯曲端之间的封闭英寸</li><li>重复步骤2.3 - 2.5余下的联系人。让尽可能紧。交替CONTA克拉导致在阵列框架的每一侧。 </li></ol> 3.袖口布局指南<ol><li>创建袖口的平开位置的二维图。 注:使用任何CAD软件来产生一个真正的大规模图。该图将确定在电极和放置位的各种电极元件的尺寸。 </li><li>打印在普通印刷纸上的二维示意图使用普通印刷机按比例绘制，然后通过5厘米见方的片与位于中央的绘图切出5厘米。 </li><li> 5厘米见方的一块热板的透明度（T1）用手术刀切出5厘米。 </li><li>放置在图纸张的顶部的透明片T1，然后将两个层在与图朝上底板。胶带下来，用胶带底板。 </li></ol> 4.电极基础层和参考联系人布局<ol><li>切出5厘米约5cm硅胶片用手术刀（S1），并且第恩将其放置在透明层上。通过丢弃一个角然后慢慢降低的片材的其余部分，以避免在T1和S1片（ 图2A）之间捕集的气泡开始。 </li><li>混合大约2g未固化的硅的指示对制造商的数据表。两部分严格搅拌用消毒的木制搅拌粘在一起。放置在真空腔室中的混合物3分钟。循环真空，因为他们浮出水面，以消除泡沫。预热烤箱isotemp在130ºC。 注意:乳胶手套可以抑制硅的固化过程。乳胶手套还含有硫可在工作表面上留下污染物。建议使用丁腈手套代替。 </li><li>使用牙科拾取工具，应用未固化的硅的薄线沿它们所在的指导图上的间隔件段的中间。 </li><li>将垫片到指定区域，然后按下来对硅胶片S1。</li><li>部分固化在isotemp烤箱硅30分钟，让其冷却10分钟。 </li><li>将参考接触到指定区域。确保焊接点朝上，接触导线对袖口出口在远端中线路由。确保正确定位后，向下按接触到硅层S1。存未固化的硅到通孔。 </li><li>磁带走在动态再完全固化硅酮以130ºC90分钟，或在室温下过夜（ 图2B）。 </li></ol> 5.中心联系阵列布局<ol><li>切出1.5厘米约5cm透明度片用手术刀（T2）。磁带走在参考从中间区域导致远，以防止它们在下一步骤中，触点阵列下方运行。 </li><li>放置触点阵列的专用位置与引线侧朝上。沉积未固化的硅到钉阵列中地点。 </li><li>广场5.1（T2）在电极的中线并在阵列中的一块保存下来，然后用胶带将两端同时按下向下阵列。手动对齐与专用位置的阵列。大盘回落袖口的边界之外的线索。 </li><li>放置在电极的中心，并通过透明段T2中的小夹具栏。以适度的压力压向基硅氧烷层S1中的中间触头夹紧它向下到基板。 </li><li>完全固化的硅氧烷为90分钟，在130ºC，或在RT过夜。 </li></ol> 6.嵌入电极组件<ol><li>取下小灯条，轻轻取出透明板T2，露出中间触点阵列。删除所有持有两个引用和中间触点（ 图2C）导线的磁带。 </li><li>切一块方形透明片材的用手术刀的相同的宽度电极和5厘米的长度（T3），然后切一块方形硅胶片的覆盖整个电极表面（S2）。 </li><li>躺在透明度片（T3）的顶部的硅胶片（S2）及拉伸它，以除去任何波或不规则性和对被截留在两者之间消除气泡。 </li><li>切四件硅胶管; 5厘米长的每个。将它们放置在作为分配的指导图上的引线的出口部位。离开电极边缘和管&#39;的边缘之间有2毫米的空间。按住每对管用镊子，大盘回落管在1毫米开始从管端走。重复其他对。 </li><li>安排和中间触点与引用成束的导线，然后通过靠近出口部位对应的管传递它们。重复其他三个管。 （ 图2D）。 </li><li>沉积在整个电极体未固化的硅的慷慨的金额。 注:避免形成在此步骤期间由任一慢慢倾倒从抽真空的混合容器中的未固化的硅树脂或用注射器注入它红外气泡。 </li><li>放置6.3结构上与硅氧烷薄片S2朝下沉积未固化的硅的顶部。与电极对准透明度片T3同时保持硅酮片材S2粘合到它。 </li><li>大盘回落的透明度件T3，然后施加压力，以引导出任何截留气泡。放置在电极的中心，并通过透明段T3中的大的夹具杆。然后夹住它下降到中度压力底板。完全固化的硅氧烷为90分钟，在130ºC，或在RT过夜。 </li></ol> 7.屏蔽层放置（推荐录音护腕） <ol><li>如果大的夹具酒吧和分层透明片（T3）用镊子。放置屏蔽片在所述电极的每一个面的中心和施加轻微的压力吨o按他们进入电极。存未固化的硅到通孔。 </li><li>部分固化的硅氧烷在130ºC30分钟，然后让它完全冷却到室温。放置胶带在电极的外端和在闭合凸缘，以防止增加额外的未固化的硅对这些段。 </li><li>重复步骤6.6至6.8。 </li></ol> 8.切出来的成品电极<ol><li>剥离并切成使用手术刀刀片在步骤7.2中加入粘合带的顶部的过量硅氧烷，然后小心地取下胶带。 </li><li>通过硅切割出窗口通过S2层，以暴露间隔段。提取用镊子嵌入间隔段。这一步会留下空隙，形成在这些地区（原来S1）灵活的单硅胶片。 </li><li>剥离上覆盖硅酮管中的粘合带的顶部的过量硅氧烷，然后用手术刀BLAD剪E要与电极体平管。 </li><li>围绕电极的外周降低至基板。 </li><li>切出每个管对之间的三角形完全穿过底板，并在外侧以下的指导图塑造引线&#39;出口点。删除从电极体中最后一个步骤超脱所有的硅材料。 </li></ol> 9.揭露联系人和屏蔽层<ol><li>通过硅树脂层S2覆盖屏蔽层切出窗口。滑行聚丙烯缝线丝在电极基底（层S1）和在基板上脱层成品袖带电极的透明层T1之间。 </li><li>翻转电极，该中心的联系和有机硅层S1朝上，再通过基地的有机硅层S1切割出窗揭露他们。重复外部参考接触暴露宽1毫米段沿c中心ontacts。确保通孔在基准触点的两侧稳定完全嵌入电极的体内。 </li></ol> 10.焊接一个连接器连接到信息<ol><li>存款焊接物质到动态分别到连接器管脚，和然后加热并用烙铁熔化两个部分在一起。 注意:DFT引线由银核由做出来的镍钴基合金MP35N的外层包围。沉积焊料物质到这些导线需要使用特种焊剂，以允许附着到导线（请参考材料清单）。 </li></ol>

<ol>
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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+stimulation+of+the+canine+hypoglossal+nerve+using+a+multi-contact+cuff+electrode.">Selective stimulation of the canine hypoglossal nerve using a multi-contact cuff electrode.</a> Ann Bio Med Eng. 32, 511-519 (2004).</li><li>Rydevik, B., Lundborg, G., Bagge, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+graded+compression+on+intraneural+blood+flow:+An+in+vivo+study+on+rabbit+tibial+nerve.">Effects of graded compression on intraneural blood flow: An in vivo study on rabbit tibial nerve.</a> J hand Surg. 6, 3-12 (1981).</li><li>Ogata, K., Naito, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+flow+of+peripheral+nerve+effects+of+dissection,+stretching+and+compression.">Blood flow of peripheral nerve effects of dissection, stretching and compression.</a> J Hand Sur. 11, 10-14 (1986).</li><li>Boretius, T., 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=A+transverse+intrafascicular+multichannel+electrode+(TIME)+to+interface+with+the+peripheral+nerve.">A transverse intrafascicular multichannel electrode (TIME) to interface with the peripheral nerve.</a> Bio Sen and Bio Elec. 26, 62-69 (2010).</li><li>Stieglitz, T., Schuettler, M., Meyer, J. U., Micromachined, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=polyimide-based+devices+for+flexible+neural+interfaces.">polyimide-based devices for flexible neural interfaces.</a> Bio Med Micro Dev. 2, 283-294 (2000).</li></ol>

作者宣称，他们没有竞争的经济利益。仅供参考在这个手稿中列出的供应商。

在这篇文章中描述的制造方法需要以确保最终箍的质量灵巧和精细运动。记录触点必须精确地在两个参考电极的中间放置。这种布局已经显示出从周围的肌肉电活动27显著减少干扰。在制造过程中的接触的相对位置的任何不平衡会降低排斥反应的压脉袋外部产生的共模干扰信号。然而，通过仔细技术很少在基准触点没有明显的不平衡进行了观察。 一些改善，到袖口的?...

与外周神经系统（PNS）的接口提供了高度处理的神经命令信号访问，因为它们前往不同结构的主体内。这些信号由分册内密闭轴突生成和由紧密接合束膜细胞包围。从神经活动产生的可测量的电势的量值，如围绕分册高电阻束膜层受神经内的各个层的阻抗。因此，两种接口方法已经依赖于相对于该束膜层，即神经束内和extrafascicular办法的记录位置被探索。内部束状的方法把各分册内的电极。这些方法的实例是犹他州阵列17，纵向帧内束状电极（LIFE）18，和横向帧内束状多通道电极（TIME）32。 ŧHESE技术可以从神经选择性地记录，但没有示出在体内 ，由于尺寸和电极12的遵守长时间，可能可靠地保留功能。 特束状办法把周围神经的触点。在这些方法中使用的压脉袋电极不损害神经束膜也不外膜，并已被证明是既从外周神经系统12记录的一个安全可靠的手段。然而，超束状方法缺乏测量单单位活动的能力 - 相比于内部束状设计。即利用神经套囊电极Neuroprosthetic应用包括下肢的激活，膀胱，隔膜，治疗慢性疼痛，神经传导阻滞，感觉反馈，并记录electroneurograms 1。潜在的应用能够利用外周神经接口包括休息O形环移动到与功能性电刺激麻痹的受害者，记录从剩余的神经运动神经元活动中被截肢者，以控制动力假肢，并与自主神经系统提供生物电子药品20接口。 一个设计实现袖口电极是平接口神经电极（FINE）21。这种设计重塑神经与较大的圆周平面的横截面相比圆形。这种设计的优点是增加了可以被放置在神经触点数目，并与选择性记录和刺激的重排的内部分册触头的附近。此外，在大型动物和人类上肢和下肢神经可以采取各种形状和由精细生成的整形不变形的神经的自然几何形状。最近的试验已经表明，细能够在恢复感觉上肢16和恢复运动中与人类功能性电刺激的下肢22。 一个箍电极的基本结构包括放置多个金属触点的神经节段的表面上，然后绝缘这些联系人用不导电袖带内的神经节段沿。为了实现这个基本结构，几种设计已经提出了在以前的研究中，包括: （1） 金属接触嵌入到的确良网格 。网状然后神经周围缠绕，将所得压脉袋的形状如下所述神经几何4,5。 （2），使用预先形刚性和非导电气缸拆分缸设计来修复神经周围的联系人。接收到该压脉袋的神经片段被整形到箍的内部几何6 - 8。 ="jove_content"&gt;（3）自卷取 ，其中触点被两绝缘层之间的封闭设计 。而与外部的未拉伸层拉伸内部层被熔化。具有不同的自然休息长度为两粘结层使最终结构以形成柔性螺旋，它包装本身周围的神经。用于这些层的材料通常都被聚9聚酰亚胺10和硅橡胶1。 （4）抵靠神经引线非绝缘部分以用作电极接触。这些线索要么编织成硅管11或硅胶套缸12成型。类似的原理，使用通过排列和融合绝缘导线以形成阵列，然后通过绝缘的开口是通过这些接合线13的中间剥一小部分制成构造的罚款。这些设计的屁股UME圆神经横截面和符合此假定神经几何。 （5） 基于弹性聚酰亚胺电极 33与由微机械加工的聚酰亚胺结构，然后整合到拉伸硅酮片材，以形成自卷绕翻边形成触点。这样的设计也假设一个圆形的神经横截面。 袖带电极应该是灵活的和自我上浆，以避免拉伸和压缩，可能会导致神经损伤3的神经。一些已知的机制，使压脉袋的电极可诱导这些影响力从邻近肌肉袖带并因此神经传输，翻边和神经的机械性能，并在压脉袋的引线过分张力之间的失配。这些安全问题导致对机械柔性组特定的设计限制，几何结构和大小1。这些标准特别challe在高接触计数精细的情况下nging因为袖口必须是在横向方向上具有相同的时间僵硬重塑神经和灵活的在纵向方向上，以防止损坏以及容纳多个联系人。自上浆螺旋设计可以容纳多个联系人袖套14，但所得到的压脉袋是稍微刚性的。柔性聚酰亚胺设计可以容纳大量的接触，但容易发生分层。导线阵列设计13产生具有扁平横截面的细，但为了保持这种几何形状的导线沿袖口生产硬表面和锋利边缘使然后不适于长期植入物的长度熔合在一起。 在这篇文章中描述的制造技术生产与可以是手工制作，精度一贯的高弹性结构的高接触密度的罚款。它采用了刚性聚合物（聚醚醚酮（PEEK）），以允许精确p触点lacement。将PEEK段在电极的中心保持扁平断面，同时保持在沿神经纵向方向柔性的。这样的设计也减小了封套的总厚度和刚度由于电极体不必是为了平坦化神经或固定触头的刚性。

<table><tbody><tr><td>Platinum-Iridium foil</td><td>Alfa Aesar</td><td>41802</td><td>90% Platinum Iridium&amp;nbsp;</td></tr><tr><td>DFT wires</td><td>Fort Wayne Metals</td><td>35N LT-DFT-28%Ag</td><td></td></tr><tr><td>Lead connector</td><td>Omnetics Connector Corporation</td><td>MCS-27-SS</td><td></td></tr><tr><td>Silicone sheet</td><td>Speciality Silicon Fabricator</td><td>0.005&quot; x 12&quot; x 12&quot; Silicone Sheet</td><td>High durometer, vulcanized&amp;nbsp;</td></tr><tr><td>Polyether ether ketone (PEEK) sheet</td><td>Peek-Optima</td><td>0.005 sheet LT3 grade</td><td></td></tr><tr><td>polyester stabelizing mesh</td><td>Surgicalmesh</td><td>PETKM2002</td><td></td></tr><tr><td>Silicon tubing (0.04&quot; I.D. 0.085&quot; O.D.)</td><td>Silcon Medical/NewAge Industries.</td><td>2810458</td><td></td></tr><tr><td>Outer shielding layer</td><td>Alfa Aesar, A Johnson Matthey</td><td>MFCD00003436 (11391)</td><td>Gold foil, 0.004&quot; thick</td></tr><tr><td>Transparency sheet</td><td>APOLLO</td><td>APOCG7060</td><td></td></tr><tr><td>Ultrasonic bath cleaner</td><td>Terra Universal</td><td>2603-00A-220</td><td></td></tr><tr><td>Isotemp standard lab oven</td><td>Fisher Scientific</td><td>13247637G</td><td></td></tr><tr><td>Optical microscope</td><td>Fisher Scientific</td><td>15-000-101</td><td></td></tr><tr><td>Tweezers</td><td>Technik</td><td>18049USA (2A-SA)</td><td></td></tr><tr><td>Surgical blade handles</td><td>Aspen Surgical Products</td><td>371031</td><td></td></tr><tr><td>Base frame&amp;nbsp;</td><td>McMaster-Carr</td><td>9785K411</td><td></td></tr><tr><td>Support beam</td><td>McMaster-Carr</td><td>9524K359</td><td></td></tr><tr><td>Two parts silicone</td><td>Nusil</td><td>MED 4765</td><td></td></tr><tr><td>Soldering Flux</td><td>SRA Soldering Products</td><td>FLS71</td><td></td></tr><tr><td>Tape</td><td>3M Healthcare</td><td>1535-0 (SKUMMM15350H)</td><td>Paper, hypoallergenic surgical tape</td></tr><tr><td>Spot welding machine</td><td>Unitek</td><td>125 Power Supply with 101F Welding Head</td><td></td></tr><tr><td>Laser cutting platform</td><td>Universal Laser Systems</td><td>PLS6.150D</td><td>150 watts laser</td></tr></tbody></table>

fabrication of high contact-density, flat-interface nerve electrodes for recording and stimulation applications

已经进行了许多尝试来制造多接触神经套囊电极是安全，健壮和可靠的长期neuroprosthetic应用程序。这个协议描述了一种改性圆柱形神经套囊电极的制造技术，以满足这些条件。最低计算机辅助设计与制造（CAD和CAM）技术是必要的，始终如一地生产高精度（接触位置0.51±0.04毫米），各种尺寸的袖口袖口。在空间上分布的接触和保留这种设计完成预定的几何形状的能力，精确度是两个重要的标准，以优化袖口的接口选择性录音和刺激。所提出的设计，可以在纵向方向最大化灵活性，同时保持足够的刚性，在横向方向上，通过使用材料具有不同的弹性重塑神经。袖口的横截面的扩张观察区域作为增加袖带内的压力的结果，为25％，在67毫米汞柱。该测试表明了箍的弹性和它的神经响应肿胀植入后。触点的稳定性"接口和记录品质也研究了与联系人的阻抗和信噪比度量从一个长期植入箍（7.5个月），并且观察到2.55±0.25分别kΩ和5.10±0.81分贝。

（ - 7 kHz带宽和2000总增益700赫兹）录制的神经活动与使用超β输入仪表放大器定制的前置放大器进行。与所提出的协议所制造的精细电极的一个例子示于图3。植入神经周围的精细由两个自由边缘缝合在一起完成的。的压脉袋的灵活性（ 图3B）的演示指示箍变平的神经，同时保留在纵向方向上的灵活性。 除了在纵向方向上的压脉袋的灵活性，袖带还?...

Watch this Scientific Journal Video about Fabrication of High Contact-Density, Flat-Interface Nerve Electrodes for Recording and Stimulation Applications at JoVE.com

Fabrication of High Contact-Density, Flat-Interface Nerve Electrodes for Recording and Stimulation Applications

本文提供了一个高接触密度扁平接口神经电极（FINE）的制造过程的详细描述。该电极用于记录和外周神经中选择性刺激神经活动优化。

高接触密度的制作，平面接口神经电极用于记录和刺激的应用

Many attempts have been made to manufacture multi-contact nerve cuff electrodes that are safe, robust and reliable for long term neuroprosthetic ...

fabrication-high-contact-density-flat-interface-nerve-electrodes-for

Research

JoVE Journal

Neuroscience

9.7K 次观看.  Case Western Reserve University. 本文提供了一个高接触密度扁平接口神经电极（FINE）的制造过程的详细描述。该电极用于记录和外周神经中选择性刺激神经活动优化。 

视频: Fabrication of High Contact-Density, Flat-Interface Nerve Electrodes for Recording and Stimulation Applications

Electrode Fabrication and Implantation in Aplysia californica for Multi-channel Neural and Muscular Recordings in Intact, Freely Behaving Animals

Recording from key nerves and muscles of Aplysia during feeding behavior allows us to study the patterns of neural control in an intact animal. Simultaneously recording from multiple nerves and muscles gives us precise information about the timing of neural activity. Previous recording methods have worked for two electrodes, but the study of additional nerves or muscles required combining and averaging the recordings of multiple animals, which made it difficult to determine fine details of timing and phasing, because of variability from response to response, and from animal to animal. Implanting four individual electrodes has a very low success rate due to the formation of adhesions that prevent animals from performing normal feeding movements. We developed a new method of electrode fabrication that reduces the bulk of the electrodes inside the animal allowing for normal feeding movements. Using a combination of glues to attach the electrodes results in a more reliable insulation of the electrode which lasts longer, making it possible to record for periods as long as a week. The fabrication technique that we describe could be extended to incorporate several additional electrodes, and would be applicable to vertebrate animals.

Electrode Fabrication and Implantation in Aplysia californica for Multi-channel Neural and Muscular Recordings in Intact, Freely Behaving Animals

A technique is described for implanting four in vivo electrodes to monitor the neuromuscular control of feeding behavior in Aplysia californica.

电极的制作及植入海兔californica完整的，自由的行为的动物多通道的神经和肌肉的录音

Recording from key nerves and muscles of Aplysia during feeding behavior allows us to study the patterns of neural control in an intact animal.  ...

科研

神经科学

Combined Shuttle-Box Training with Electrophysiological Cortex Recording and Stimulation as a Tool to Study Perception and Learning

Shuttle-box avoidance learning is a well-established method in behavioral neuroscience and experimental setups were traditionally custom-made; the necessary equipment is now available by several commercial companies. This protocol provides a detailed description of a two-way shuttle-box avoidance learning paradigm in rodents (here Mongolian gerbils; Meriones unguiculatus) in combination with site-specific electrical intracortical microstimulation (ICMS) and simultaneous chronical electrophysiological in vivo recordings. The detailed protocol is applicable to study multiple aspects of learning behavior and perception in different rodent species.
Site-specific ICMS of auditory cortical circuits as conditioned stimuli here is used as a tool to test the perceptual relevance of specific afferent, efferent and intracortical connections. Distinct activation patterns can be evoked by using different stimulation electrode arrays for local, layer-dependent ICMS or distant ICMS sites. Utilizing behavioral signal detection analysis it can be determined which stimulation strategy is most effective for eliciting a behaviorally detectable and salient signal. Further, parallel multichannel-recordings using different electrode designs (surface electrodes, depth electrodes, etc.) allow for investigating neuronal observables over the time course of such learning processes. It will be discussed how changes of the behavioral design can increase the cognitive complexity (e.g. detection, discrimination, reversal learning).

Shuttle-box avoidance learning is well-established in behavioral neuroscience. This protocol describes how shuttle-box learning in rodents can be combined with site-specific electrical intracortical microstimulation (ICMS) and simultaneous chronical in vivo recordings as a tool to study multiple aspects of learning and perception.

联合穿梭盒子训练与电的Cortex记录和刺激作为一种工具来研究感知和学习

Shuttle-box avoidance learning is a well-established method in behavioral neuroscience and experimental setups were traditionally custom-made; the ...

行为学

Fabrication of Ti3C2 MXene Microelectrode Arrays for In Vivo Neural Recording

Implantable microelectrode technologies have been widely used to elucidate neural dynamics at the microscale to gain a deeper understanding of the neural underpinnings of brain disease and injury. As electrodes are miniaturized to the scale of individual cells, a corresponding rise in the interface impedance limits the quality of recorded signals. Additionally, conventional electrode materials are stiff, resulting in a significant mechanical mismatch between the electrode and the surrounding brain tissue, which elicits an inflammatory response that eventually leads to a degradation of the device performance. To address these challenges, we have developed a process to fabricate flexible microelectrodes based on Ti3C2 MXene, a recently discovered nanomaterial that possesses remarkably high volumetric capacitance, electrical conductivity, surface functionality, and processability in aqueous dispersions. Flexible arrays of Ti3C2 MXene microelectrodes have remarkably low impedance due to the high conductivity and high specific surface area of the Ti3C2 MXene films, and they have proven to be exquisitely sensitive for recording neuronal activity. In this protocol, we describe a novel method for micropatterning Ti3C2 MXene into microelectrode arrays on flexible polymeric substrates and outline their use for in vivo micro-electrocorticography recording. This method can easily be extended to create MXene electrode arrays of arbitrary size or geometry for a range of other applications in bioelectronics and it can also be adapted for use with other conductive inks besides Ti3C2 MXene. This protocol enables simple and scalable fabrication of microelectrodes from solution-based conductive inks, and specifically allows harnessing the unique properties of hydrophilic Ti3C2 MXene to overcome many of the barriers that have long hindered the widespread adoption of carbon-based nanomaterials for high-fidelity neural microelectrodes.

Fabrication of Ti3C2 MXene Microelectrode Arrays for In Vivo Neural Recording

We describe here a method for fabricating Ti3C2 MXene microelectrode arrays and utilizing them for in vivo neural recording.

用于Vivo神经记录的 Ti3C2 MXene 微电极阵列的制造

Implantable microelectrode technologies have been widely used to elucidate neural dynamics at the microscale to gain a deeper understanding of the neural ...

生物工程

Fabrication of the Composite Regenerative Peripheral Nerve Interface (C-RPNI) in the Adult Rat

Recent advances in neuroprosthetics have enabled those living with extremity loss to reproduce many functions native to the absent extremity, and this is often accomplished through integration with the peripheral nervous system. Unfortunately, methods currently employed are often associated with significant tissue damage which prevents prolonged use. Additionally, these devices often lack any meaningful degree of sensory feedback as their complex construction dampens any vibrations or other sensations a user may have previously depended on when using more simple prosthetics. The composite regenerative peripheral nerve interface (C-RPNI) was developed as a stable, biologic construct with the ability to amplify efferent motor nerve signals while providing simultaneous afferent sensory feedback. The C-RPNI consists of a segment of free dermal and muscle graft secured around a target mixed sensorimotor nerve, with preferential motor nerve reinnervation of the muscle graft and sensory nerve reinnervation of the dermal graft. In rats, this construct has demonstrated the generation of compound muscle action potentials (CMAPs), amplifying the target nerve&#39;s signal from the micro- to milli-volt level, with signal to noise ratios averaging approximately 30-50. Stimulation of the dermal component of the construct generates compound sensory nerve action potentials (CSNAPs) at the proximal nerve. As such, this construct has promising future utility towards the realization of the ideal, intuitive prosthetic.

Fabrication of the Composite Regenerative Peripheral Nerve Interface C-RPNI in the Adult Rat

The following manuscript describes a novel method for developing a biologic, closed loop neural feedback system termed the composite regenerative peripheral nerve interface (C-RPNI). This construct has the ability to integrate with peripheral nerves to amplify efferent motor signals while simultaneously providing afferent sensory feedback.

成年大鼠复合再生外周神经接口（C-RPNI）的制造

Recent advances in neuroprosthetics have enabled those living with extremity loss to reproduce many functions native to the absent extremity, and this is ...

Open-source Toolkit: Benchtop Carbon Fiber Microelectrode Array for Nerve Recording

Conventional peripheral nerve probes are primarily fabricated in a cleanroom, requiring the use of multiple expensive and highly specialized tools. This paper presents a cleanroom "light" fabrication process of carbon fiber neural electrode arrays that can be learned quickly by an inexperienced cleanroom user. This carbon fiber electrode array fabrication process requires just one cleanroom tool, a Parylene C deposition machine, that can be learned quickly or outsourced to a commercial processing facility at marginal cost. This fabrication process also includes hand-populating printed circuit boards, insulation, and tip optimization. 
The three different tip optimizations explored here (Nd:YAG laser, blowtorch, and UV laser) result in a range of tip geometries and 1 kHz impedances, with blowtorched fibers resulting in the lowest impedance. While previous experiments have proven laser and blowtorch electrode efficacy, this paper also shows that UV laser-cut fibers can record neural signals in vivo. Existing carbon fiber arrays either do not have individuated electrodes in favor of bundles or require cleanroom fabricated guides for population and insulation. The proposed arrays use only tools that can be used at a benchtop for fiber population. This carbon fiber electrode array fabrication process allows for quick customization of bulk array fabrication at a reduced price compared to commercially available probes.

Here, we describe fabrication methodology for customizable carbon fiber electrode arrays for recording in vivo in nerve and brain.

开源工具包:用于神经记录的台式碳纤维微电极阵列

Conventional peripheral nerve probes are primarily fabricated in a cleanroom, requiring the use of multiple expensive and highly specialized tools. This ...

Chronic Implantation of Multiple Flexible Polymer Electrode Arrays

Simultaneous recordings from large populations of individual neurons across distributed brain regions over months to years will enable new avenues of scientific and clinical development. The use of flexible polymer electrode arrays can support long-lasting recording, but the same mechanical properties that allow for longevity of recording make multiple insertions and integration into a chronic implant a challenge. Here is a methodology by which multiple polymer electrode arrays can be targeted to a relatively spatially unconstrained set of brain areas.
The method utilizes thin-film polymer devices, selected for their biocompatibility and capability to achieve long-term and stable&#160;electrophysiologic recording interfaces. The resultant implant allows accurate and flexible targeting of anatomically distant regions, physical stability for months, and robustness to electrical noise. The methodology supports up to sixteen serially inserted devices across eight different anatomic targets. As previously demonstrated, the methodology is capable of recording from 1024 channels. Of these, the 512 channels in this demonstration used for single neuron recording yielded 375 single units distributed across six recording sites. Importantly, this method also can record single units for at least 160 days.
This implantation strategy, including temporarily bracing each device with a retractable silicon insertion shuttle, involves tethering of devices at their target depths to a skull-adhered plastic base piece that is custom-designed for each set of recording targets, and stabilization/protection of the devices within a silicone-filled, custom-designed plastic case. Also covered is the preparation of devices for implantation, and design principles that should guide adaptation to different combinations of brain areas or array designs.

Described below is a method for implantation of multiple polymer electrode arrays across anatomically distant brain regions for chronic electrophysiological recording in freely moving rats. Preparation and surgical implantation are described in detail, with emphasis on design principles to guide adaptation of these methods for use in other species.

多柔性聚合物电极阵列的慢性植入

Simultaneous recordings from large populations of individual neurons across distributed brain regions over months to years will enable new avenues of ...

Preparation of Peripheral Nerve Stimulation Electrodes for Chronic Implantation in Rats

Peripheral nerve cuff electrodes have long been used in the neurosciences and related fields for stimulation of, for example, vagus or sciatic nerves. Several recent studies have demonstrated the effectiveness of chronic VNS in enhancing central nervous system plasticity to improve motor rehabilitation, extinction learning, and sensory discrimination. Construction of chronically implantable devices for use in such studies is challenging due to rats&#8217; small size, and typical protocols require extensive training of personnel and time-consuming microfabrication methods. Alternatively, commercially available implantable cuff electrodes can be purchased at a significantly higher cost. In this protocol, we present a simple, low-cost method for construction of small, chronically implantable peripheral nerve cuff electrodes for use in rats. We validate the short and long-term reliability of our cuff electrodes by demonstrating that VNS in ketamine/xylazine anesthetized rats produces decreases in breathing rate consistent with activation of the Hering-Breuer reflex, both at the time of implantation and up to 10 weeks after device implantation. We further demonstrate the suitability of the cuff electrodes for use in chronic stimulation studies by pairing VNS with skilled lever press performance to induce motor cortical map plasticity.

Existing approaches for constructing chronically implantable peripheral nerve cuff electrodes for use in small rodents often require specialized equipment and/or highly trained personnel. In this protocol we demonstrate a simple, low-cost approach for fabricating chronically implantable cuff electrodes, and demonstrate their effectiveness for vagus nerve stimulation (VNS) in rats.

大鼠慢性植入的外周神经刺激电极的制备

Peripheral nerve cuff electrodes have long been used in the neurosciences and related fields for stimulation of, for example, vagus or sciatic nerves. ...

Construction and Implementation of Carbon Fiber Microelectrode Arrays for Chronic and Acute In Vivo Recordings

Multichannel electrode arrays offer insight into the working brain and serve to elucidate neural processes at the single-cell and circuit levels. Development of these tools is crucial for understanding complex behaviors and cognition and for advancing clinical applications. However, it remains a challenge to densely record from cell populations stably and continuously over long time periods. Many popular electrodes, such as tetrodes and silicon arrays, feature large cross-diameters that produce damage upon insertion and elicit chronic reactive tissue responses associated with neuronal death, hindering the recording of stable, continuous neural activity. In addition, most wire bundles exhibit broad spacing between channels, precluding simultaneous recording from a large number of cells clustered in a small area. The carbon fiber microelectrode arrays described in this protocol offer an accessible solution to these concerns. The study provides a detailed method for fabricating carbon fiber microelectrode arrays that can be used for both acute and chronic recordings in vivo. The physical properties of these electrodes make them ideal for stable and continuous long-term recordings at high cell densities, enabling the researcher to make robust, unambiguous recordings from single units across months.

Construction and Implementation of Carbon Fiber Microelectrode Arrays for Chronic and Acute In Vivo Recordings

This protocol describes a procedure for constructing carbon fiber microelectrode arrays for chronic and acute in vivo electrophysiological recordings in mouse (Mus musculus) and ferret (Mustela putorius furo) from multiple brain regions. Each step, following the purchase of raw carbon fibers to microelectrode array implantation, is described in detail, with emphasis on microelectrode array construction.

用于慢性和急性体内记录的碳纤维微电极阵列 的 构建和实现

Multichannel electrode arrays offer insight into the working brain and serve to elucidate neural processes at the single-cell and circuit levels. ...

Preparation of Rat Sciatic Nerve for Ex Vivo Neurophysiology

Ex vivo preparations enable the study of many neurophysiological processes in isolation from the rest of the body while preserving local tissue structure. This work describes the preparation of rat sciatic nerves for ex vivo neurophysiology, including buffer preparation, animal procedures, equipment setup and neurophysiological recording. This work provides an overview of the different types of experiments possible with this method. The outlined method aims to provide 6 h of stimulation and recording on extracted peripheral nerve tissue in tightly controlled conditions for optimal consistency in results. Results obtained using this method are A-fibre compound action potentials (CAP) with peak-to-peak amplitudes in the millivolt range over the entire duration of the experiment. CAP amplitudes and shapes are consistent and reliable, making them useful to test and compare new electrodes to existing models, or the effects of interventions on the tissue, such as the use of chemicals, surgical alterations, or neuromodulatory stimulation techniques. Both conventional commercially available cuff electrodes with platinum-iridium contacts and custom-made conductive elastomer electrodes were tested and gave similar results in terms of nerve stimulus strength-duration response.

Preparation of Rat Sciatic Nerve for Ex Vivo Neurophysiology

This protocol describes the preparation of rat whole sciatic nerve tissue for ex vivo electrophysiological stimulation and recording in an environmentally-regulated, two-compartment, perfused saline bath.

大鼠坐骨神经用于 离体 神经生理学的制备

Ex vivo preparations enable the study of many neurophysiological processes in isolation from the rest of the body while preserving local tissue structure. ...

<meta charset="utf8"></head><body> 为临床前啮齿动物模型制造肌外肌电图电极

Development of a Low-cost Epimysial Electromyography Electrode: A Simplified Workflow for Fabrication and Testing

Electromyography (EMG) is a valuable diagnostic tool for detecting neuromuscular abnormalities. Implantable epimysial electrodes are commonly used to measure EMG signals in preclinical models. Although classical resources exist describing the principles of epimysial electrode fabrication, there is a sparsity of illustrative information translating electrode theory to practice. To remedy this, we provide an updated, easy-to-follow guide on fabricating and testing a low-cost epimysial electrode. 
Electrodes were made by folding and inserting two platinum-iridium foils into a precut silicone base to form the contact surfaces. Next, coated stainless steel wires were welded to each contact surface to form the electrode leads. Lastly, a silicone mixture was used to seal the electrode. Ex vivo testing was conducted to compare our custom-fabricated electrode to an industry standard electrode in a saline bath, where high levels of signal agreement (sine [intraclass correlation - ICC= 0.993], square [ICC = 0.995], triangle [ICC = 0.958]), and temporal-synchrony (sine [r = 0.987], square [r = 0.990], triangle [r= 0.931]) were found across all waveforms. Low levels of electrode impedance were also quantified via electrochemical impedance spectroscopy. 
An in vivo performance assessment was also conducted where the vastus lateralis muscle of a rat was surgically instrumented with the custom-fabricated electrode and signaling was acquired during uphill and downhill walking. As expected, peak EMG activity was significantly lower during downhill walking (0.008 &plusmn; 0.005 mV) than uphill (0.031 &plusmn; 0.180 mV, p = 0.005), supporting the validity of the device. The reliability and biocompatibility of the device were also supported by consistent signaling during level walking at 14 days and 56 days post implantation (0.01 &plusmn; 0.007 mV, 0.012 &plusmn; 0.007 mV respectively; p &gt; 0.05) and the absence of histological inflammation. Collectively, we provide an updated workflow for the fabrication and testing of low-cost epimysial electrodes.

<meta charset="utf8"></head><body>作者聚焦:ACL 损伤研究中的肌外膜电极制造和测试

Our purpose was to provide an updated, easy-to-follow guide on the fabrication and testing of epimysial electromyography electrodes. To that end, we provide instructions for material sourcing and a detailed walkthrough of the fabrication and testing process.

低成本肌外肌电图电极的开发:简化的制造和测试工作流程

Electromyography (EMG) is a valuable diagnostic tool for detecting neuromuscular abnormalities. Implantable epimysial electrodes are commonly used to ...

高接触密度的制作，平面接口神经电极用于记录和刺激的应用

摘要

探索更多视频

电极的制作及植入海兔californica完整的，自由的行为的动物多通道的神经和肌肉的录音

联合穿梭盒子训练与电的Cortex记录和刺激作为一种工具来研究感知和学习

用于Vivo神经记录的 Ti₃C₂ MXene 微电极阵列的制造

成年大鼠复合再生外周神经接口（C-RPNI）的制造

开源工具包:用于神经记录的台式碳纤维微电极阵列

多柔性聚合物电极阵列的慢性植入

大鼠慢性植入的外周神经刺激电极的制备

用于慢性和急性体内记录的碳纤维微电极阵列的构建和实现

大鼠坐骨神经用于离体神经生理学的制备

低成本肌外肌电图电极的开发:简化的制造和测试工作流程

高接触密度的制作，平面接口神经电极用于记录和刺激的应用

摘要

探索更多视频

电极的制作及植入海兔californica完整的，自由的行为的动物多通道的神经和肌肉的录音

联合穿梭盒子训练与电的Cortex记录和刺激作为一种工具来研究感知和学习

用于Vivo神经记录的 Ti3C2 MXene 微电极阵列的制造

成年大鼠复合再生外周神经接口（C-RPNI）的制造

开源工具包:用于神经记录的台式碳纤维微电极阵列

多柔性聚合物电极阵列的慢性植入

大鼠慢性植入的外周神经刺激电极的制备

用于慢性和急性体内记录的碳纤维微电极阵列 的 构建和实现

大鼠坐骨神经用于 离体 神经生理学的制备

低成本肌外肌电图电极的开发:简化的制造和测试工作流程

用于Vivo神经记录的 Ti₃C₂ MXene 微电极阵列的制造

用于慢性和急性体内记录的碳纤维微电极阵列的构建和实现

大鼠坐骨神经用于离体神经生理学的制备