这项工作是由神经通信技术中心（CNCT），P41资源中心，由国家生物医学成像和生物工程研究所（NIBIB，P41 EB002030）和国立卫生研究院（NIH）支持。作者希望感谢NeuroNexus技术维特尔里约热内卢的外科技术的发展，段密歇根大学编制在手术中使用的设备清单帕特尔。

1。植入前程序（共同） <ol><li>刮胡子动物的头顶部之间的眼睛以及耳朵后面。使用电动理发推剪，剃了头。头先用异丙醇擦洗，然后优碘（重复3倍）。应用眼睛润滑油。 </li><li>装载动物的立体耳酒吧。动物的右耳外耳道直接进入移动对运河侧着脑袋正确的立体耳酒吧固定的小费。必须采取非常谨慎，以确保每个耳朵酒吧的小费是正确定位内耳道。接下来，推入外耳道松动的左耳栏，并适用于轻微的外来压力。掌握动物的鼻子和摆动来回坚决。如果动物是正确安装的头会觉得安装牢固。 </li></ol> 2。预先植入手术（共同） <ol><li>一双细剪刀，中国保监会切两半从中线向外LES，部分消除头皮</li><li>凑和删除坚持骨骨膜的结缔组织。如手术刀刀片或消毒棉签的尖端背面的边缘钝刮，将最大限度地减少出血，因为毛细血管会被压碎，而不是削减顺利。控制骨出血用骨蜡或烧灼的铁。 </li><li>使用双氧水消毒和清洁的骨面。干骨，使颅缝线更清晰可见。前囟门和Lambda点应注意，用平头。开颅手术和电极植入位置，应测量前囟门就可以暂时用钢笔或由浅到表面钻标记。开颅的位置将取决于研究的实验目的（即电机，感官实验）1。 </li><li>钻入骨screwholes，并放置在地面和稳定小号船员。请务必使用不锈钢或钛螺钉。珠宝商的小螺丝（例如＃2-56，＃00-80），与尽可能多线程每英寸是可取的。 </li></ol> 3。硅电极植入程序<ol><li>钻孔后植入螺钉的开颅手术。开颅角落钻四个浅的飞行员马克。使用试点标记为指导，深入开颅手术的周长。钻孔用一双坚固镊子取出骨中心质量。开放开颅手术的边缘，可能需要进一步钻探清洗。与microrongeurs对薄骨片可能会被裁剪。精心计划你的手术，以避免大血管，如矢状窦，可能出现在0.5毫米的中线纵向缝合1。矢状窦穿刺可引起广泛出血。暴露硬脑膜应保持水合学联或生理盐水。 </li><li>刺破硬脑膜您将需要使用无菌注射器针头的小计（例如，&lt;28计）时尚小钩。以针锥，并针对商号表面按平（如手术刀的平坦部分）稍倾斜，形成一个90度的弯点。用这个钩子来捕获硬脑膜表面，解除脑表面向上。硬脑膜可以减少使用微型剪刀或使用的硬脑膜钩的横向运动可能被撕毁。照顾，以避免大脑表面的血管使用硬脑膜挂钩时。你应该注意到脑脊液漏时，被切除硬脑膜和硬脑膜表面的差异在色彩。 </li><li>小心地连接地线接地螺钉基地周围的硅电极，机智与牙科水泥的地方。所以应放在仔细硅电极，电极电缆引线圆拱了开颅手术。 </li><li>植入物将放在采用线性电机驱动器的解放军海关在立体定向仪。附加电极插入设备，聚乙二醇（PEG）是用烙铁加热和应用插入杆和记录电极。 </li><li>电极保护，手动降低电极的尖端到所需的大脑表面的立体位置。 </li><li>使用的软件接口，线性驱动器，移动到所需深度的脑组织内的电极。 </li><li>大脑中的安全记录电极，电极电缆连接相邻的螺丝PEG。现在，您可以加入生理盐水溶解聚乙二醇必将电极插入设备中删除。提高插入设备远离了开颅手术，并为每个电极的重复。 </li><li>用盐水浸泡明胶海绵覆盖暴露的开颅手术。有机硅聚合物，然后用牙科丙烯酸覆盖电极电缆和保护。在决赛中的连接器的位置位置，然后申请牙科丙烯酸强大headcap。 </li></ol> 4。 MicroECoG电极植入程序<ol><li>的microECoG电极植入将涉及稍大的开颅手术。为5×5毫米的设备，一个6×6毫米的开颅手术必须进行。钻前的开颅手术，牙科紫外光固化丙烯酸的开颅手术现场的外围，而它仍然是干燥和不接触硬脑膜或PIA的危险。 </li><li> ＃107毛刺用一个无菌手术钻钻骨的开颅手术的一般形状的表面。较小＃106毛刺可以被用来钻出来其余的骨薄薄的透明层。 rongeurs届时能够解除关闭剩余的骨片。硬膜外或硬膜下可植入的microECoG。再次，保持硬脑膜与人工脑脊液或生理盐水水合。 </li><li>在为了植入microECoG的，将OPE立体手臂列印头骨，并确保使用无菌磁带手臂的电极。电极，现在可以降低到开颅手术。确保电极站点正面临着向下，将硬膜或软膜表面的接触。如果保持湿润，电极应滑入组织，将留在原地。 </li><li>接地线连接到接地螺钉环绕着下至少三次。接地螺钉可以是任何螺丝与硬脑膜接触。参考线，也可以以同样的方式连接到参考螺丝。 </li><li>盐水浸泡明胶海绵小块应放置电极周围哪里有硬膜或软暴露。少量的盐水浸泡明胶海绵，还应该包括薄膜电极的顶部。可应用于牙科紫外光固化丙烯酸明胶海绵顶部，可以用来建立一个稳定的头。直接薄膜电缆coveri的丙烯酸应用吴它，直到连接达到。要小心，不要覆盖以外的连接器盖的底部。 </li><li>另外，植入在4.3 microECoG的，可以紧接着植入了硅微电极阵列通过制造在microECoG基板的小孔（步骤3.2-3.6）。这使得单单位同时在不同深度，连同周围的脑表面的高分辨率现场录音录音。 </li></ol> 5。术后恢复<ol><li>水泥完全硬化后，缝合皮肤紧紧围绕头帽和从动物的立体框架。 </li><li>适用于防腐粉末丰富地在伤口周围或抗生素软膏。如果有任何从耳朵出血，放入耳道一些抗生素良好。 </li><li>保持温暖的动物，如巴比妥类药物防止动物维持体内热量。楼选管会Ĥ长期植入在一个单独的笼子里的动物。它通常需要4至7天，动物完全从手术中恢复。 </li></ol> 6。代表结果一个成功的硅电极植入手术将录制与之间500kOhm-2MOhm测量阻抗的网站，和薄膜电极之间10kOhm-50kOhm（1kHz时）。神经信号可以检查以及手术后立即。你应该能够看到在植入电极的尖峰和薄膜表面电极（图1）上看到的慢波振荡。 <img src="/files/ftp_upload/3565/3565fig1.jpg" alt="图1" /> 图1。单单位活动和脑电图振荡代表性的结果。6通道同步记录的一种慢性硅电极植入大脑皮层一）宽带数据。请注意，经常可以看到在记录点的尖峰。 b）电子从6通道16通道薄膜uECoG大脑表面的电极阵列COG振荡。所有16个信号的平均值分别从每个跟踪（常见的平均引用），并应用了500赫兹数字低通滤波器。

<ol>
	<li>Paxinos, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Rat Brain in Stereotaxic Coordinates. , (2007).</li><li>Fujisawa, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavior-dependent+short-term+assembly+dynamics+in+the+medial+prefrontal+cortex.">Behavior-dependent short-term assembly dynamics in the medial prefrontal cortex.</a> Nat. Neurosci. 11 (7), 823-833 (2008).</li><li>Merrill, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrical+stimulation+of+excitable+tissue:+design+of+efficacious+and+safe+protocols.">Electrical stimulation of excitable tissue: design of efficacious and safe protocols.</a> J. Neurosci. Methods. 141, 171-171 (2005).</li><li>Gage, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Naive+coadaptive+cortical+control.">Naive coadaptive cortical control.</a> J. Neural Eng. 2, 52-52 (2005).</li><li>Marzullo, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+Closed-Loop+Neural+Interface+Technology+in+a+Rat+Model:+Combining+Motor+Cortex+Operant+Conditioning+With+Visual+Cortex+Microstimulation.">Development of Closed-Loop Neural Interface Technology in a Rat Model: Combining Motor Cortex Operant Conditioning With Visual Cortex Microstimulation.</a> IEEE Trans. Neur. Sys. and Rehab. Eng. 18 (2), 117-126 (2010).</li><li>Maynard, E. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Utah+intracortical+electrode+array:+a+recording+structure+for+potential+brain-computer+interfaces.">The Utah intracortical electrode array: a recording structure for potential brain-computer interfaces.</a> Electroencephalogr. Clin. Neurophysiol. 102, 228-239 (1997).</li><li>Kipke, D. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Silicon-substrate+intracortical+microelectrode+arrays+for+long-term+recording+of+neuronal+spike+activity+in+cerebral+cortex.">Silicon-substrate intracortical microelectrode arrays for long-term recording of neuronal spike activity in cerebral cortex.</a> IEEE Trans. Neural Syst. Rehab. Eng. 11 (2), 151-155 (2003).</li><li>Vetter, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+neural+recording+using+silicon-substrate+microelectrode+arrays+implanted+in+cerebral+cortex.">Chronic neural recording using silicon-substrate microelectrode arrays implanted in cerebral cortex.</a> IEEE Trans. Biomed. Eng. 51 (6), 896-904 (2004).</li></ol>

DRK是对NeuroNexus科技的部分业主。

有越来越大的兴趣在研究脑功能2，提供微刺激，或控制信号4,5为神经义肢系统使用皮质内和表面的多声道录音接口。在这部影片中所述的方法演示了如何植入慢性穿透性和表面电极系统。而其他慢性硅电极系统存在6，我们一直致力于开发由NeuroNexus科技7,8的平面电极植入。植入电极系统的技术将有很大的不同。 所描述的手术步骤，安装在立体耳酒吧动物，可能是最困难和最重要的一步，在确定植入电极位置的准确性。因此，必须格外小心，假设正确地定位内耳道每个耳朵酒吧的小费。后的AssuraNCE的耳朵，酒吧小费是在动物的右耳朵的听觉鼻道，释放你的右手动物的脖子上，而你的左拇指和食指继续以支持头部和适用于对在耳，酒吧一角头部的压力。用你的右手，推入外耳道松动的左耳栏，并适用于轻微的外来压力。动物的头部左侧，向下和向前和各地移动，直到你能感觉到左耳扎入内耳道尖滑。用你的右手，继续适用于左耳栏上的压力。释放动物的头，你现在可以自由的左手，拧紧固定螺钉的左耳栏。

<table border="1"><tbody><tr><td> 设备 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>骨螺钉 - ＃2-56×3/16，十字盘头，18-8的S / </td><td> BoltDepot </td><td> 7650 </td><td></td></tr><tr><td>骨螺钉 - ＃2-56×1/8，十字盘头，18-8的S / </td><td> BoltDepot </td><td> 9617 </td><td></td></tr><tr><td>大氧气罐 - 大小为k </td><td>低温气体</td><td> OXKME </td><td></td></tr><tr><td>小氧气罐 - 大小é </td><td>低温气体</td><td> OXYE-AL </td><td></td></tr><tr><td> vannas蒂宾根弹簧剪刀 - 2.5毫米刀片，斜角</td><td>罚款科学工具</td><td> 15002-08 </td><td></td></tr><tr><td>杜蒙＃5镊子，特氟隆</td><td>罚款科学工具</td><td> 11626-11 </td><td></td></tr><tr><td>杜蒙＃5镊子，Dumostar </td><td>罚款科学工具</td><td> 11295-10 </td><td></td></tr><tr><td>学生Vannas春季剪刀</td><td>罚款科学工具</td><td> 91500-09 </td><td></td></tr><tr><td> adson钳</td><td>罚款科学工具</td><td> 11006-12 </td><td></td></tr><tr><td> friendman-皮尔森Ronqeur，弯曲</td><td>罚款科学工具</td><td> 16221-14 </td><td></td></tr><tr><td>虹膜剪刀 - ToughCut </td><td>罚款科学工具</td><td> 14058-11 </td><td></td></tr><tr><td> vannas蒂宾根弹簧剪刀 - 5mm的刀片，直</td><td>连接NE科学工具</td><td> 15003-08 </td><td></td></tr><tr><td>手术刀手柄</td><td>罚款科学工具</td><td> 10003-12 </td><td></td></tr><tr><td>塑料消毒箱</td><td>罚款科学工具</td><td> 20810-02 </td><td></td></tr><tr><td>不锈钢钢尺</td><td>罚款科学工具</td><td> 30086-15 </td><td></td></tr><tr><td>探头和抹刀 - 直14厘米</td><td>罚款科学工具</td><td> 10090-13 </td><td></td></tr><tr><td>止血钳直</td><td>罚款科学工具</td><td> 13002-10 </td><td></td></tr><tr><td>止血钳弯</td><td>罚款科学工具</td><td> 13003-10 </td><td></td></tr><tr valign="bottom"><td>微钻头站inless钢锉 - 0.7毫米</td><td>罚款科学工具</td><td> 19008-07 </td><td> 10/Pack </td></tr><tr valign="bottom"><td>微钻头不锈钢毛刺 - 0.5毫米</td><td>罚款科学工具</td><td> 19007-05 </td><td> 10/Pack </td></tr><tr valign="bottom"><td>骨螺钉，自攻 - 1.17毫米（直径） - 4.7毫米（长） </td><td>罚款科学工具</td><td> 19010-00 </td><td> 100/Pack </td></tr><tr valign="bottom"><td>人工泪液</td><td>费舍尔</td><td> NC9860842 </td><td>每</td></tr><tr valign="bottom"><td>优碘</td><td>费舍尔</td><td> 19-066452 </td><td>瓶</td></tr><tr valign="bottom"><td>的F-AIR活性炭过滤器</td><td>费舍尔</td><td> NC9112250 </td><td>罐</td></tr><tr valign="bottom"><td>撒施棉花通风报信，非无菌</td> &lt;TD&gt;物资服务</td><td> 1104 </td><td> 10Pack/Case </td></tr><tr valign="bottom"><td>纱布2"×2" </td><td>物资服务</td><td> 1630 </td><td> 25Pack/Case </td></tr><tr valign="bottom"><td>针尖，23遗传x 1" </td><td>物资服务</td><td> 39412 </td><td>销售由案例</td></tr><tr valign="bottom"><td>针尖，27遗传×1.25" </td><td>物资服务</td><td> 25251 </td><td></td></tr><tr valign="bottom"><td>针尖，30遗传×0.5" </td><td>物资服务</td><td> 22023 </td><td> 10Box/Case </td></tr><tr valign="bottom"><td>软膏，三联抗生素0.6克</td><td>物资服务</td><td> 2528 </td><td></td></tr><tr valign="bottom"><td>袋，自密封灭菌Pouche<html> ，3.5"×8" </td><td>物资服务</td><td> 1023 </td><td> 10Box/Case </td></tr><tr valign="bottom"><td>铃声解决方案千毫升</td><td>物资服务</td><td> 5263 </td><td> 12Bag/Case </td></tr><tr valign="bottom"><td>手术刀刀片一次性使用无菌＃15 </td><td>物资服务</td><td> 1975年</td><td> 3Box/Case </td></tr><tr valign="bottom"><td>缝合，的尼龙w/P-3针，5-0，18英寸。 </td><td>物资服务</td><td> 4618 </td><td> 12/Box </td></tr><tr valign="bottom"><td> underpad，一次性23"×36" </td><td>物资服务</td><td> 2545 </td><td> 15Pack/Case </td></tr><tr valign="bottom"><td>润滑剂，果冻外科无菌</td><td>物资服务</td><td> 2538 </td><td> 12Box/Case </td></tr>"&gt; <td>吸收泡沫凝胶2cmx6cmx7mm </td><td>物资服务</td><td> 3107 </td><td></td></tr><tr valign="bottom"><td>六件组螺丝起子组</td><td>国家珠宝用品</td><td> ETSCR98001 </td><td></td></tr><tr valign="bottom"><td> KWIK银胶</td><td> WPI的</td><td> KWIK-SIL </td><td></td></tr><tr valign="bottom"><td> KWIK银管</td><td> WPI的</td><td> 600009 </td><td></td></tr><tr valign="top"><td>慢性硅电极16到64个站点</td><td> NeuroNexus技术</td><td>会因探针设计</td><td></td></tr><tr valign="top"><td>慢性薄膜表面电极（microECoG）16至32个站点</td><td> NeuroNexus技术</td><td>会因探针设计</td><td>&nbsp;
 </td>
 </tr>
 </tbody>
</table>

surgical implantation of chronic neural electrodes for recording single unit activity and electrocorticographic signals

长期的电生理记录的成功往往取决于植入手术的质量。在这里，我们谁是学习的过程中植入电极系统的外科医生提供有用的信息。我们展示的渗透和表面电极植入手术。手术过程描述，从开始到结束，包括整个过程的每一步的详细说明。还应当指出，这个视频指南，重点对啮齿动物模型和其他小动物模型中进行的程序。所描述的程序修改为其他的动物模型是可行的。

Watch this Scientific Journal Video about Surgical Implantation of Chronic Neural Electrodes for Recording Single Unit Activity and Electrocorticographic Signals at JoVE.com

Surgical Implantation of Chronic Neural Electrodes for Recording Single Unit Activity and Electrocorticographic Signals

我们提供了学习慢性神经记录电极植入过程中的外科医生，谁是有用的信息。啮齿动物模型中描述的穿透和表面电极系统的技术。

用于记录单机活动和Electrocorticographic信号的慢性神经电极植入手术

The success of long-term electrophysiological recordings often depends on the quality of the implantation surgery.  Here we provide useful information for ...

surgical-implantation-chronic-neural-electrodes-for-recording-single

Research

JoVE Journal

Neuroscience

47.2K 次观看.  University of Michigan. 我们提供了学习慢性神经记录电极植入过程中的外科医生，谁是有用的信息。啮齿动物模型中描述的穿透和表面电极系统的技术。

视频: Surgical Implantation of Chronic Neural Electrodes for Recording Single Unit Activity and Electrocorticographic Signals

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from individual neurons are critical for understanding how neural circuits function under natural conditions. Traditionally, these recordings have been performed 'blind', meaning the identity of the recorded cell is unknown at the start of the recording. Cellular identity can be subsequently determined via intracellular1, juxtacellular2 or loose-patch3 iontophoresis of dye, but these recordings cannot be pre-targeted to specific neurons in regions with functionally heterogeneous cell types. Fluorescent proteins can be expressed in a cell-type specific manner permitting visually-guided single-cell electrophysiology4-6. However, there are many model systems for which these genetic tools are not available. Even in genetically accessible model systems, the desired promoter may be unknown or genetically homogenous neurons may have varying projection patterns. Similarly, viral vectors have been used to label specific subgroups of projection neurons7, but use of this method is limited by toxicity and lack of trans-synaptic specificity. Thus, additional techniques that offer specific pre-visualization to record from identified single neurons in vivo are needed. Pre-visualization of the target neuron is particularly useful for challenging recording conditions, for which classical single-cell recordings are often prohibitively difficult8-11. The novel technique described in this paper uses retrograde transport of a fluorescent dye applied using tungsten needles to rapidly and selectively label a specific subset of cells within a particular brain region based on their unique axonal projections, thereby providing a visual cue to obtain targeted electrophysiological recordings from identified neurons in an intact circuit within a vertebrate CNS.
The most significant novel advancement of our method is the use of fluorescent labeling to target specific cell types in a non-genetically accessible model system. Weakly electric fish are an excellent model system for studying neural circuits in awake, behaving animals12. We utilized this technique to study sensory processing by "small cells" in the anterior exterolateral nucleus (ELa) of weakly electric mormyrid fish. "Small cells" are hypothesized to be time comparator neurons important for detecting submillisecond differences in the arrival times of presynaptic spikes13. However, anatomical features such as dense myelin, engulfing synapses, and small cell bodies have made it extremely difficult to record from these cells using traditional methods11, 14. Here we demonstrate that our novel method selectively labels these cells in 28% of preparations, allowing for reliable, robust recordings and characterization of responses to electrosensory stimulation.

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograde transport of fluorescent dye labels a sub-population of neurons based on anatomical projection. Labeled axons can be visually targeted in vivo, permitting extracellular recording from identified axons. This technique facilitates recording when neurons cannot be labeled through genetic manipulation or are difficult to isolate using 'blind' in vivo approaches.

逆行荧光染料标记允许有针对性的外单细胞记录技术鉴定神经元<em&gt;在体内</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

科研

神经科学

Recording Human Electrocorticographic (ECoG) Signals for Neuroscientific Research and Real-time Functional Cortical Mapping

Neuroimaging studies of human cognitive, sensory, and motor processes are usually based on noninvasive techniques such as electroencephalography (EEG), magnetoencephalography or functional magnetic-resonance imaging. These techniques have either inherently low temporal or low spatial resolution, and suffer from low signal-to-noise ratio and/or poor high-frequency sensitivity. Thus, they are suboptimal for exploring the short-lived spatio-temporal dynamics of many of the underlying brain processes. In contrast, the invasive technique of electrocorticography (ECoG) provides brain signals that have an exceptionally high signal-to-noise ratio, less susceptibility to artifacts than EEG, and a high spatial and temporal resolution (i.e., &lt;1 cm/&lt;1 millisecond, respectively). ECoG involves measurement of electrical brain signals using electrodes that are implanted subdurally on the surface of the brain. Recent studies have shown that ECoG amplitudes in certain frequency bands carry substantial information about task-related activity, such as motor execution and planning1, auditory processing2 and visual-spatial attention3. Most of this information is captured in the high gamma range (around 70-110 Hz). Thus, gamma activity has been proposed as a robust and general indicator of local cortical function1-5. ECoG can also reveal functional connectivity and resolve finer task-related spatial-temporal dynamics, thereby advancing our understanding of large-scale cortical processes. It has especially proven useful for advancing brain-computer interfacing (BCI) technology for decoding a user's intentions to enhance or improve communication6 and control7. Nevertheless, human ECoG data are often hard to obtain because of the risks and limitations of the invasive procedures involved, and the need to record within the constraints of clinical settings. Still, clinical monitoring to localize epileptic foci offers a unique and valuable opportunity to collect human ECoG data. We describe our methods for collecting recording ECoG, and demonstrate how to use these signals for important real-time applications such as clinical mapping and brain-computer interfacing. Our example uses the BCI2000 software platform8,9 and the SIGFRIED10 method, an application for real-time mapping of brain functions. This procedure yields information that clinicians can subsequently use to guide the complex and laborious process of functional mapping by electrical stimulation.
Prerequisites and Planning:
Patients with drug-resistant partial epilepsy may be candidates for resective surgery of an epileptic focus to minimize the frequency of seizures. Prior to resection, the patients undergo monitoring using subdural electrodes for two purposes: first, to localize the epileptic focus, and second, to identify nearby critical brain areas (i.e., eloquent cortex) where resection could result in long-term functional deficits. To implant electrodes, a craniotomy is performed to open the skull. Then, electrode grids and/or strips are placed on the cortex, usually beneath the dura. A typical grid has a set of 8 x 8 platinum-iridium electrodes of 4 mm diameter (2.3 mm exposed surface) embedded in silicon with an inter-electrode distance of 1cm. A strip typically contains 4 or 6 such electrodes in a single line. The locations for these grids/strips are planned by a team of neurologists and neurosurgeons, and are based on previous EEG monitoring, on a structural MRI of the patient's brain, and on relevant factors of the patient's history. Continuous recording over a period of 5-12 days serves to localize epileptic foci, and electrical stimulation via the implanted electrodes allows clinicians to map eloquent cortex. At the end of the monitoring period, explantation of the electrodes and therapeutic resection are performed together in one procedure.
In addition to its primary clinical purpose, invasive monitoring also provides a unique opportunity to acquire human ECoG data for neuroscientific research. The decision to include a prospective patient in the research is based on the planned location of their electrodes, on the patient's performance scores on neuropsychological assessments, and on their informed consent, which is predicated on their understanding that participation in research is optional and is not related to their treatment. As with all research involving human subjects, the research protocol must be approved by the hospital's institutional review board. The decision to perform individual experimental tasks is made day-by-day, and is contingent on the patient's endurance and willingness to participate. Some or all of the experiments may be prevented by problems with the clinical state of the patient, such as post-operative facial swelling, temporary aphasia, frequent seizures, post-ictal fatigue and confusion, and more general pain or discomfort.
At the Epilepsy Monitoring Unit at Albany Medical Center in Albany, New York, clinical monitoring is implemented around the clock using a 192-channel Nihon-Kohden Neurofax monitoring system. Research recordings are made in collaboration with the Wadsworth Center of the New York State Department of Health in Albany. Signals from the ECoG electrodes are fed simultaneously to the research and the clinical systems via splitter connectors. To ensure that the clinical and research systems do not interfere with each other, the two systems typically use separate grounds. In fact, an epidural strip of electrodes is sometimes implanted to provide a ground for the clinical system. Whether research or clinical recording system, the grounding electrode is chosen to be distant from the predicted epileptic focus and from cortical areas of interest for the research. Our research system consists of eight synchronized 16-channel g.USBamp amplifier/digitizer units (g.tec, Graz, Austria). These were chosen because they are safety-rated and FDA-approved for invasive recordings, they have a very low noise-floor in the high-frequency range in which the signals of interest are found, and they come with an SDK that allows them to be integrated with custom-written research software. In order to capture the high-gamma signal accurately, we acquire signals at 1200Hz sampling rate-considerably higher than that of the typical EEG experiment or that of many clinical monitoring systems. A built-in low-pass filter automatically prevents aliasing of signals higher than the digitizer can capture. The patient's eye gaze is tracked using a monitor with a built-in Tobii T-60 eye-tracking system (Tobii Tech., Stockholm, Sweden). Additional accessories such as joystick, bluetooth Wiimote (Nintendo Co.), data-glove (5th Dimension Technologies), keyboard, microphone, headphones, or video camera are connected depending on the requirements of the particular experiment.
Data collection, stimulus presentation, synchronization with the different input/output accessories, and real-time analysis and visualization are accomplished using our BCI2000 software8,9. BCI2000 is a freely available general-purpose software system for real-time biosignal data acquisition, processing and feedback. It includes an array of pre-built modules that can be flexibly configured for many different purposes, and that can be extended by researchers' own code in C++, MATLAB or Python. BCI2000 consists of four modules that communicate with each other via a network-capable protocol: a Source module that handles the acquisition of brain signals from one of 19 different hardware systems from different manufacturers; a Signal Processing module that extracts relevant ECoG features and translates them into output signals; an Application module that delivers stimuli and feedback to the subject; and the Operator module that provides a graphical interface to the investigator.
A number of different experiments may be conducted with any given patient. The priority of experiments will be determined by the location of the particular patient's electrodes. However, we usually begin our experimentation using the SIGFRIED (SIGnal modeling For Realtime Identification and Event Detection) mapping method, which detects and displays significant task-related activity in real time. The resulting functional map allows us to further tailor subsequent experimental protocols and may also prove as a useful starting point for traditional mapping by electrocortical stimulation (ECS).
Although ECS mapping remains the gold standard for predicting the clinical outcome of resection, the process of ECS mapping is time consuming and also has other problems, such as after-discharges or seizures. Thus, a passive functional mapping technique may prove valuable in providing an initial estimate of the locus of eloquent cortex, which may then be confirmed and refined by ECS. The results from our passive SIGFRIED mapping technique have been shown to exhibit substantial concurrence with the results derived using ECS mapping10.
The protocol described in this paper establishes a general methodology for gathering human ECoG data, before proceeding to illustrate how experiments can be initiated using the BCI2000 software platform. Finally, as a specific example, we describe how to perform passive functional mapping using the BCI2000-based SIGFRIED system.

Recording Human Electrocorticographic ECoG Signals for Neuroscientific Research and Real-time Functional Cortical Mapping

We present a method for collecting electrocorticographic signals for research purposes from humans who are undergoing invasive epilepsy monitoring. We show how to use the BCI2000 software platform for data collection, signal processing and stimulus presentation. Specifically, we demonstrate SIGFRIED, a BCI2000-based tool for real-time functional brain mapping.

神经科学的研究和实时功能皮层映射记录人力Electrocorticographic（ECOG）信号

Neuroimaging studies of human cognitive, sensory, and motor processes are usually based on noninvasive techniques such as electroencephalography (EEG), ...

Optical Recording of Suprathreshold Neural Activity with Single-cell and Single-spike Resolution

Signaling of information in the vertebrate central nervous system is often carried by populations of neurons rather than individual neurons. Also propagation of suprathreshold spiking activity involves populations of neurons. Empirical studies addressing cortical function directly thus require recordings from populations of neurons with high resolution. Here we describe an optical method and a deconvolution algorithm to record neural activity from up to 100 neurons with single-cell and single-spike resolution. This method relies on detection of the transient increases in intracellular somatic calcium concentration associated with suprathreshold electrical spikes (action potentials) in cortical neurons. High temporal resolution of the optical recordings is achieved by a fast random-access scanning technique using acousto-optical deflectors (AODs)1. Two-photon excitation of the calcium-sensitive dye results in high spatial resolution in opaque brain tissue2. Reconstruction of spikes from the fluorescence calcium recordings is achieved by a maximum-likelihood method. Simultaneous electrophysiological and optical recordings indicate that our method reliably detects spikes (&gt;97% spike detection efficiency), has a low rate of false positive spike detection (&lt; 0.003 spikes/sec), and a high temporal precision (about 3 msec) 3. This optical method of spike detection can be used to record neural activity in vitro and in anesthetized animals in vivo3,4.

Understanding the function of the vertebrate central nervous system requires recordings from many neurons because cortical function arises on the level of populations of neurons. Here we describe an optical method to record suprathreshold neural activity with single-cell and single-spike resolution, dithered random-access scanning. This method records somatic fluorescence calcium signals from up to 100 neurons with high temporal resolution. A maximum-likelihood algorithm deconvolves the underlying suprathreshold neural activity from the somatic fluorescence calcium signals. This method reliably detects spikes with high detection efficiency and a low rate of false positives and can be used to study neural populations in vitro and in vivo.

阈上的神经活动的单细胞和单穗的分辨率的光学的方法记录

Signaling of information in the vertebrate central nervous system is often carried by populations of neurons rather than individual neurons. Also ...

Fiber-optic Implantation for Chronic Optogenetic Stimulation of Brain Tissue

Elucidating patterns of neuronal connectivity has been a challenge for both clinical and basic neuroscience. Electrophysiology has been the gold standard for analyzing patterns of synaptic connectivity, but paired electrophysiological recordings can be both cumbersome and experimentally limiting. The development of optogenetics has introduced an elegant method to stimulate neurons and circuits, both in vitro1 and in vivo2,3. By exploiting cell-type specific promoter activity to drive opsin expression in discrete neuronal populations, one can precisely stimulate genetically defined neuronal subtypes in distinct circuits4-6. Well described methods to stimulate neurons, including electrical stimulation and/or pharmacological manipulations, are often cell-type indiscriminate, invasive, and can damage surrounding tissues. These limitations could alter normal synaptic function and/or circuit behavior. In addition, due to the nature of the manipulation, the current methods are often acute and terminal. Optogenetics affords the ability to stimulate neurons in a relatively innocuous manner, and in genetically targeted neurons. The majority of studies involving in vivo optogenetics currently use a optical fiber guided through an implanted cannula6,7; however, limitations of this method include damaged brain tissue with repeated insertion of an optical fiber, and potential breakage of the fiber inside the cannula. Given the burgeoning field of optogenetics, a more reliable method of chronic stimulation is necessary to facilitate long-term studies with minimal collateral tissue damage. Here we provide our modified protocol as a video article to complement the method effectively and elegantly described in Sparta et al.8 for the fabrication of a fiber optic implant and its permanent fixation onto the cranium of anesthetized mice, as well as the assembly of the fiber optic coupler connecting the implant to a light source. The implant, connected with optical fibers to a solid-state laser, allows for an efficient method to chronically photostimulate functional neuronal circuitry with less tissue damage9 using small, detachable, tethers. Permanent fixation of the fiber optic implants provides consistent, long-term in vivo optogenetic studies of neuronal circuits in awake, behaving mice10 with minimal tissue damage.

The development of optogenetics now provides the means to precisely stimulate genetically defined neurons and circuits, both in vitro and in vivo. Here we describe the assembly and implantation of a fiber optic for chronic photostimulation of brain tissue.

光纤植入慢性Optogenetic刺激脑组织

Elucidating patterns of neuronal connectivity has been a challenge for both clinical and basic neuroscience. Electrophysiology has been the gold standard ...

Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

Detection and interpretation of olfactory cues are critical for the survival of many organisms. Remarkably, species across phyla have strikingly similar olfactory systems suggesting that the biological approach to chemical sensing has been optimized over evolutionary time1. In the insect olfactory system, odorants are transduced by olfactory receptor neurons (ORN) in the antenna, which convert chemical stimuli into trains of action potentials. Sensory input from the ORNs is then relayed to the antennal lobe (AL; a structure analogous to the vertebrate olfactory bulb). In the AL, neural representations for odors take the form of spatiotemporal firing patterns distributed across ensembles of principal neurons (PNs; also referred to as projection neurons)2,3. The AL output is subsequently processed by Kenyon cells (KCs) in the downstream mushroom body (MB), a structure associated with olfactory memory and learning4,5. Here, we present electrophysiological recording techniques to monitor odor-evoked neural responses in these olfactory circuits.
First, we present a single sensillum recording method to study odor-evoked responses at the level of populations of ORNs6,7. We discuss the use of saline filled sharpened glass pipettes as electrodes to extracellularly monitor ORN responses. Next, we present a method to extracellularly monitor PN responses using a commercial 16-channel electrode3. A similar approach using a custom-made 8-channel twisted wire tetrode is demonstrated for Kenyon cell recordings8. We provide details of our experimental setup and present representative recording traces for each of these techniques.

Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

We demonstrate variations of the extracellular multi-unit recording technique to characterize odor-evoked responses in the first three stages of the invertebrate olfactory pathway. These techniques can easily be adapted to examine ensemble activity in other neural systems as well.

多单位记录方法来描述蝗虫的神经活动（<em&gt; Schistocerca美洲</em&gt;）嗅觉电路

Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

A Method for Systematic Electrochemical and Electrophysiological Evaluation of Neural Recording Electrodes

New materials and designs for neural implants are typically tested separately, with a demonstration of performance but without reference to other implant characteristics.&#160;This precludes a rational selection of a particular implant as optimal for a particular application and the development of new materials based on the most critical performance parameters.&#160;This article develops a protocol for in vitro and in vivo testing of neural recording electrodes.&#160;Recommended parameters for electrochemical and electrophysiological testing are documented with the key steps and potential issues discussed.&#160;This method eliminates or reduces the impact of many systematic errors present in simpler in vivo testing paradigms, especially variations in electrode/neuron distance and between animal models.&#160;The result is a strong correlation between the critical in vitro and in vivo responses, such as impedance and signal-to-noise ratio.&#160;This protocol can easily be adapted to test other electrode materials and designs.&#160;The in vitro techniques can be expanded to any other nondestructive method to determine further important performance indicators.&#160;The principles used for the surgical approach in the auditory pathway can also be modified to other neural regions or tissue.

Different electrode coatings affect neural recording performance through changes to electrochemical, chemical and mechanical properties. Comparison of electrodes in vitro is relatively simple, however comparison of in vivo response is typically complicated by variations in electrode/neuron distance and between animals. This article provides a robust method to compare neural recording electrodes.

对于系统电化学神经记录电极和电生理评价方法

New materials and designs for neural implants are typically tested separately, with a demonstration of performance but without reference to other implant ...

A Guide to In vivo Single-unit Recording from Optogenetically Identified Cortical Inhibitory Interneurons

A major challenge in neurophysiology has been to characterize the response properties and function of the numerous inhibitory cell types in the cerebral cortex. We here share our strategy for obtaining stable, well-isolated single-unit recordings from identified inhibitory interneurons in the anesthetized mouse cortex using a method developed by Lima and colleagues1. Recordings are performed in mice expressing Channelrhodopsin-2 (ChR2) in specific neuronal subpopulations. Members of the population are identified by their response to a brief flash of blue light. This technique &#8211; termed &#8220;PINP&#8221;, or Photostimulation-assisted Identification of Neuronal Populations &#8211; can be implemented with standard extracellular recording equipment. It can serve as an inexpensive and accessible alternative to calcium imaging or visually-guided patching, for the purpose of targeting extracellular recordings to genetically-identified cells. Here we provide a set of guidelines for optimizing the method in everyday practice. We refined our strategy specifically for targeting parvalbumin-positive (PV+) cells, but have found that it works for other interneuron types as well, such as somatostatin-expressing (SOM+) and calretinin-expressing (CR+) interneurons.

A Guide to In vivo Single-unit Recording from Optogenetically Identified Cortical Inhibitory Interneurons

Here we describe our strategy for obtaining stable, well-isolated single-unit recordings from identified inhibitory interneurons in the anesthetized mouse cortex. Neurons expressing ChR2 are identified by their response to blue light. The method uses standard extracellular recording equipment, and serves as an inexpensive alternative to calcium imaging or visually-guided patching.

指南<em&gt;在体内</em&gt;单单元从Optogenetically鉴定皮质抑制性录像

A major challenge in neurophysiology has been to characterize the response properties and function of the numerous inhibitory cell types in the cerebral ...

Extracellular Recording of Neuronal Activity Combined with Microiontophoretic Application of Neuroactive Substances in Awake Mice

Differences in the activity of neurotransmitters and neuromodulators, and consequently different neural responses, can be found between anesthetized and awake animals. Therefore, methods allowing the manipulation of synaptic systems in awake animals are required in order to determine the contribution of synaptic inputs to neuronal processing unaffected by anesthetics. Here, we present methodology for the construction of electrodes to simultaneously record extracellular neural activity and release multiple neuroactive substances at the vicinity of the recording sites in awake mice. By combining these procedures, we performed microiontophoretic injections of gabazine to selectively block GABAA receptors in neurons of the inferior colliculus of head-restrained mice. Gabazine successfully modified neural response properties such as the frequency response area and stimulus-specific adaptation. Thus, we demonstrate that our methods are suitable for recording single-unit activity and for dissecting the role of specific neurotransmitter receptors in auditory processing.
The main limitation of the described procedure is the relatively short recording time (~3 hr), which is determined by the level of habituation of the animal to the recording sessions. On the other hand, multiple recording sessions can be performed in the same animal. The advantage of this technique over other experimental procedures used to manipulate the level of neurotransmission or neuromodulation (such as systemic injections or the use of optogenetic models), is that the drug effect is confined to the local synaptic inputs to the target neuron. In addition, the custom-manufacture of electrodes allows adjustment of specific parameters according to the neural structure and type of neuron of interest (such as the tip resistance for improving the signal-to-noise ratio of the recordings).

We present methods for the construction of electrodes to simultaneously record extracellular neural activity and release multiple neuroactive substances at the vicinity of the recording sites in awake mice. This technique allows the detailed analysis of putative local synaptic inputs to the neuron of interest.

神经元电活动外记录与神经活性物质在清醒小鼠微电泳应用相结合

Differences in the activity of neurotransmitters and neuromodulators, and consequently different neural responses, can be found between anesthetized and ...

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

Many attempts have been made to manufacture multi-contact nerve cuff electrodes that are safe, robust and reliable for long term neuroprosthetic applications. This protocol describes a fabrication technique of a modified cylindrical nerve cuff electrode to meet these criteria. Minimum computer-aided design and manufacturing (CAD and CAM) skills are necessary to consistently produce cuffs with high precision (contact placement 0.51 &#177; 0.04 mm) and various cuff sizes. The precision in spatially distributing the contacts and the ability to retain a predefined geometry accomplished with this design are two criteria essential to optimize the cuff&#39;s interface for selective recording and stimulation. The presented design also maximizes the flexibility in the longitudinal direction while maintaining sufficient rigidity in the transverse direction to reshape the nerve by using materials with different elasticities. The expansion of the cuff&#39;s cross sectional area as a result of increasing the pressure inside the cuff was observed to be 25% at 67 mm Hg. This test demonstrates the flexibility of the cuff and its response to nerve swelling post-implant. The stability of the contacts&#39; interface and recording quality were also examined with contacts&#39; impedance and signal-to-noise ratio metrics from a chronically implanted cuff (7.5 months), and observed to be 2.55 &#177; 0.25 k&#8486; and 5.10 &#177; 0.81 dB respectively.

This article provides a detailed description on the fabrication process of a high contact-density flat interface nerve electrode (FINE). This electrode is optimized for recording and stimulating neural activity selectively within peripheral nerves.

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 ...

Implantation of Chronic Silicon Probes and Recording of Hippocampal Place Cells in an Enriched Treadmill Apparatus

An important requisite for understanding brain function is the identification of behavior and cell activity correlates. Silicon probes are advanced electrodes for large-scale electrophysiological recording of neuronal activity, but the procedures for their chronic implantation are still underdeveloped. The activity of hippocampal place cells is known to correlate with an animal&#39;s position in the environment, but the underlying mechanisms are still unclear. To investigate place cells, here we describe a set of techniques which range from the fabrication of devices for chronic silicon probe implants to the monitoring of place field activity in a cue-enriched treadmill apparatus. A micro-drive and a hat are built by fitting and fastening together 3D-printed plastic parts. A silicon probe is mounted on the micro-drive, cleaned, and coated with dye. A first surgery is performed to fix the hat on the skull of a mouse. Small landmarks are fabricated and attached to the belt of a treadmill. The mouse is trained to run head-fixed on the treadmill. A second surgery is performed to implant the silicon probe in the hippocampus, following which broadband electrophysiological signals are recorded. Finally, the silicon probe is recovered and cleaned for reuse. The analysis of place cell activity in the treadmill reveals a diversity of place field mechanisms, outlining the benefit of the approach.

We describe the diverse steps to implant chronic silicon probes and to record place cells in mice that are running head-fixed on a cue-enriched treadmill apparatus.

富氧运动器械中慢性硅探针的植入及海马位细胞的记录

An important requisite for understanding brain function is the identification of behavior and cell activity correlates. Silicon probes are advanced ...