Name: brain source imaging in preclinical rat models of focal epilepsy using high-resolution eeg recordings
Uploaded: 2015-06-06T14:00:00.000+00:00
Duration: 8 min 20 s
Description: Watch this Scientific Journal Video about Brain Source Imaging in Preclinical Rat Models of Focal Epilepsy using High-Resolution EEG Recordings at JoVE.com

作者要感谢佩德罗A.巴尔德斯埃尔南德斯，弗朗索瓦Tadel和劳埃德·史密斯提出宝贵的意见和富有成果的讨论。我们也感谢拉斐尔托雷斯的校对。

伦理声明:所有实验以下的机构动物护理和使用委员会（IACUC）在佛罗里达国际大学（IACUC 13-004）确定的政策执行。 1.脑电图记录<ol><li>脑电图迷你帽的制备<ol><li>沉浸脑电图迷你帽至少12小时在蒸馏水中0.2％的氯化钠的电极尖端。在蒸馏水轻轻冲洗脑电图迷你帽。干燥的盖和在空气中的电极。 </li><li>混合脑电图电极糊在2体积比例的0.9％NaCl溶液:1.添加一滴亚甲蓝的，这将有助于可视化的电极浆料内的电极和在皮肤上。以混合浆料中的注射器。确保有没有气泡在注射器。注入凝胶到每个32电极，填充它们，而不会引入任何气泡。建议从底部而不是顶部注入。这提供了更好的ACCESS到每个电极，并降低了凝胶的蔓延的可能性。 </li><li>打开EEG和生理记录系统，并在计算机上使用打开相应的记录软件。 </li></ol></li><li>动物准备及麻醉 注:慢性癫痫是使用协议FCD 8 Wistar大鼠产生。 （ - 400克8周龄，300）脑电图记录是在成年大鼠进行。 <ol><li>记录大鼠的体重在实验片。使用该信息来计算剂量的镇静剂（dexdomitor 0.25毫克/千克）。在用5％异氟醚和100％的氧气（1升/分，在14.7 psi）的大鼠诱导麻醉。 </li><li>修剪后大鼠的头，减少异氟烷至2％，脑电图迷你盖的整个设置过程中保持它。检查大鼠反射消失（趾捏）。通过固定用耳棒耳道放置在加热垫上在立体定位仪的鼠。确保麻醉鼻锥是安全的。 </li><li>美联社来往润滑剂眼药膏每只眼睛。 </li><li>刮胡子使用剃刀鼠头部和耳朵额外的头发。避免剃须过程中的任何出血。 注:在皮肤上留下会产生噪音，在脑电图记录的任何毛发。擦大鼠皮肤用90％的异丙醇，刺激血管和皮肤脱脂。 </li><li>将头皮上的盐水擦拭，并完全覆盖它来保持良好的皮肤电导直到脑电图迷你上限是准备放置。 </li><li>连接温度，呼吸，和三个导联心电图探头。注意，该温度是由直肠探头测量。连续监视的大鼠的生理过程中记录的程序。确保正常温度为37℃，呼吸范围是30 - 每分钟60次呼吸和心脏速率是约350 - 每分钟450次。 </li></ol></li><li>录音程序<ol><li>取出盐水擦拭对大鼠的头皮，然后将准备好的微型脑电图-CAP在它的皮肤。修复迷你帽用橡皮筋。放置一个橡胶带在头皮上的前侧，通常是在眼睛的前面，而另一个带在耳朵和颈部之间的头皮的背面。使用颈下一个塑料保护，以促进正常的呼吸。 </li><li>放一层高电导电极糊的同时在地面和参考电极。把他们放在各自的耳朵。 注:参考电极可可能放置在其它位置。 </li><li>连接脑电图迷你帽放大器和观察工作台电极阻抗的预览。检查所有电极的性能。对于高品质录音，保证阻抗值是5的范围内 - 30千欧。如果有任何嘈杂电极，提供通过移动他们的脚手架内朝向头皮或轻轻地从电极的顶部注入更多的凝胶与头皮接触更好。 </li><li>辖dexdomitor（0.25毫克/千克）intraperitoneally并立即降低异氟烷率为0％。如果呼吸率不在30 - 每分钟为60次呼吸，轻轻地开始增加异氟醚率。不超过1％，异氟醚的价值。仔细监测这一步骤，因为异氟醚和dexdomitor的混合物可能会促使动物病情危重。 注:在局灶性癫痫的临床前模型，异氟醚影响简易爆炸装置，而dexdomitor没有。异氟醚下有科目致痫较弱财产， 也就是说，相对较少的简易爆炸装置可以相对于其他条件7,14进行检测。该dexdomitor剂量是有效约2小时。因此，为了节省时间，其效果，进行制备在异氟烷。 </li><li>进行脑电图记录。在记录之后，通过脑电图迷你帽之前插入它们内部的彩色笔被去除标记在皮肤上的顶部的脑电图迷你帽的三个突出的圆的位置上。用它们作为MRI共地标-registration。就拿老鼠头的标志性建筑的照片。将大鼠背部的笼子里面和监控，直到从dexdomitor的效果完全恢复。 注意:在这个实验中，红色（对手颜色为绿色）被用来从电极位置（绿色）区分。然而，建议使用其他颜色（紫色/绿色）如小出血斑的皮肤中观察到。 </li></ol></li></ol><img alt="图1" src="/files/ftp_upload/52700/52700fig1.jpg" /> 图1.脑电图迷你帽放置在一个特定的大鼠的图片。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52700/52700fig1highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 2.脑源成像<ol><li> IED分类 注:使用MATLAB中自主开发的代码基础上，该装置检测和分级进行的先前的研究[15]。该软件将可根据用户要求。 <ol><li>通过目视检查脑电图示踪剂丢弃嘈杂的渠道。除去使用自动方法用于周期性波形减法，它是基于一个模板和相关分析的EKG工件。 注意:通常情况下，谁记录脑电图实验者共享书面试验片为基于所述阻抗值中的观察到的不良的信道信息。软件删除EKG文物也将可根据用户要求。 <img alt="图2" src="/files/ftp_upload/52700/52700fig2.jpg" /> 图2.脑电图显示跟踪不同类型的智能电子设备的一个例子。红色框表示一种类型的简易爆炸装置。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52700/52700fig2highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> </li><li>应用的带通滤波器中的3截止频率 - 150赫兹和切口过滤以除去线频率（60赫兹一般，在有些国家50 Hz）分量脱机。 </li><li>检测两种类型的简易爆炸装置（尖峰和尖波）。尖峰和尖波构成的20个大型电器事件 - 70毫秒和70 - 200分别在毫秒时间。因此，施加各自的带通滤波器（15截止频率- 50赫兹的尖峰和5 - 15赫兹为尖锐波）后，所述智能电子设备是基于振幅阈值15检测。 注:该阈值被自动设置为4σ如在先前的研究中对多组活动15建议。这里，σ是带通滤波信号的估计的标准偏差，σ=中间值{|滤波后的信号| / 0.6745}。 </li><li>子分类尖峰和尖波成不同的集群。不同的尖峰和尖波的显着特点是利用小波变换15提取。它们分分类使用k均值为多个集群，和最优簇数k用剪影来确定。 </li><li>同一群集内平均的子分类的信号。对于各IED子类型的平均EEG信号将被用于脑源的分析。 </li></ol></li><li>卷导体模型 注:以下章节中，开源软件，头脑风暴12，将与MRI地图集Wistar大鼠9中。然而，个别大鼠的MRI还可以用于如果可用来产生体积导体模型。该地图集MRI 9可以在下载<a href="http://www.idac.tohoku.ac.jp/bir/en/db/rb/101028.html">http://www.idac.tohoku.ac.jp/bir/en/</a> 。该网站提供的地图册作为NIFTI格式"的Wistar大鼠MRI地图集"部分下，它可以是注册后访问。所需的前处理的软件也可以发现在这个网站。 <ol><li>输入MRI和脑表面到软件12。 <img alt="视觉1" srC ="/文件/ ftp_upload / 52700 / 52700vis1.jpg"/&gt; <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52700/52700vis1highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> </li><li>产生头表面采用默认设置。 <img alt="视觉2" src="/files/ftp_upload/52700/52700vis2.jpg" /> <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52700/52700vis2highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> </li><li>生成基于MRI铅场计算12头皮和内/外表面的头骨。 注:顶点的分辨率影响估计源的精确度，但大量的顶点导致高计算复杂度。每一层的顶点数量，建议642是可接受的准确性与公平的计算复杂度。头骨的厚度可从MRI检查，并且在MRI图谱的情况下，它是约1μm米在软件中插入上述值，相应的三角脸顶点网格后，每个表面将被创建。 <img alt="视觉3" src="/files/ftp_upload/52700/52700vis3.jpg" /> <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52700/52700vis3highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> </li><li>检查每个表面的取向和位置使用可视化选项对于MRI检查。进行相应的修改，如果任何表面不共配准12。 <img alt="视觉4" height="351" src="/files/ftp_upload/52700/52700vis4.jpg" width="321" /> <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52700/52700vis4highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> </li><li>在使用1.3.5收购老鼠头像。共登记的3界标（R1，R2和R3）在MRI的位置。使用的地标网格点作为参考来克enerate的电极位置，电极被固定在支架上（ 图3B）。 <img alt="图3" src="/files/ftp_upload/52700/52700fig3.jpg" /> 图3.（A）老鼠头像用来获取电极位置和（b）脑电图迷你帽图的坐标系。红点在（A）表示在1.3.5中提到的地标。对应于红色的数字中的（B）。另外，在绿色标记（A）中描述了32电极位置，并且它们对应于蓝色的数字中的（B）。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52700/52700fig3highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> </li><li>基于所述3界标为N×3电极位置矩阵。这里，N是信道数（N = 32）和列表示相应的x，y和z坐标值。 注:脑电图迷你帽是刚性支架。因此，一旦3参考网格（R1，R2和R3）都得到，电极的位置被自动设定。用户将仅需要重新定义上的方式使微型帽适当地投射在头皮的Z值。如在图3B蓝色数字描绘了N点网格，可以按顺序编号。该标准支架的脑电图迷你帽可购（ 表材料 ）。该软件为共同注册也可到社区。 </li><li>输入所产生的信道的文件。 <img alt="视觉5" src="/files/ftp_upload/52700/52700vis5.jpg" /> <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52700/52700vis5highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> </li><li>显示并确认所有电极的位置。修改任何错位电极12。最终的坐标电系统德位置应与用于上述表面的坐标系一致。 注:创建的表面可以通过使用可视化选项，进行目视检查上的MRI然后，选定的表面将被显示为在磁共振成像黄线"MRI检查登记检查MRI /表面登记"。此外，3地标和32的电极位置可以通过选择工具箱的选择，显示在MRI"查看传感器磁共振查看器。"的位置可以通过基于大鼠的眼睛和耳朵的位置进行比较的分布（进行目视检查图4）。 <img alt="图4" src="/files/ftp_upload/52700/52700fig4.jpg" /> 图4.（A）的磁共振图谱与共同注册脑表面（黄线），（B）和对准32的电极和第3界标（红点）创建的卷导体模型，和（C）的核磁共振图谱与共同配准REF<html> erence电网R1。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52700/52700fig4highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> </li></ol></li><li>脑源成像<ol><li>计算铅场矩阵13。输入满足皮肤，颅骨和脑的比率为1的电导率值:1/80:1.获得基于体积导体模型和2.2产生的电极的位置的导线场矩阵。 注:该工具箱12提供的接口与其他软件计算BEM 10。因此，只有电导率值是必需作为输入。 <img alt="6视觉" src="/files/ftp_upload/52700/52700vis6.jpg" /> <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52700/52700vis6highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> </li><li>输入每个IED子类型存储在2.1.4平均EEG信号。 "SRC ="/文件/ ftp_upload / 52700 / 52700vis7.jpg"/&gt; <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52700/52700vis7highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> </li><li>获取基于所计算的铅场矩阵和输入EEG信号sLORETA溶液13。通过选择源估计方法选项，逆溶液可以得到12。 <img alt="视觉8" src="/files/ftp_upload/52700/52700vis8.jpg" /> <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52700/52700vis8highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> </li><li>绘制估计来源。 </li></ol></li></ol>

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R., Nadasdy, Z., Ben-Shaul, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unsupervised+spike+detection+and+sorting+with+wavelets+and+super-paramagnetic+clustering.">Unsupervised spike detection and sorting with wavelets and super-paramagnetic clustering.</a> Neural Computation. 16 (8), 1661-1687 (2004).</li><li>Delorme, A., Makeig, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=EEGLAB:+an+open+source+toolbox+for+analysis+of+single-trial+EEG+dynamics+including+independent+component+analysis.">EEGLAB: an open source toolbox for analysis of single-trial EEG dynamics including independent component analysis.</a> Journal of Neuroscience Methods. 134 (1), 9-21 (2004).</li><li>Oostenveld, R., Fries, P., Maris, E., Schoffelen, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FieldTrip:+Open+Source+Software+for+Advanced+Analysis+of+MEG,+EEG,+and+Invasive+Electrophysiological+Data.">FieldTrip: Open Source Software for Advanced Analysis of MEG, EEG, and Invasive Electrophysiological Data.</a> Computational Intelligence and Neuroscience. 2011, 1-9 (2011).</li><li>Koessler, L., 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=Source+localization+of+ictal+epileptic+activity+investigated+by+high+resolution+EEG+and+validated+by+SEEG.">Source localization of ictal epileptic activity investigated by high resolution EEG and validated by SEEG.</a> NeuroImage. 51 (2), 642-653 (2010).</li><li>Manganotti, P., 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=Scalp+topography+and+source+analysis+of+interictal+spontaneous+spikes+and+evoked+spikes+by+digital+stimulation+in+benign+rolandic+epilepsy.">Scalp topography and source analysis of interictal spontaneous spikes and evoked spikes by digital stimulation in benign rolandic epilepsy.</a> Electroencephalography and Clinical Neurophysiology. 107 (1), 18-26 (1998).</li><li>Bae, J., Deshmukh, A., Song, Y., Brain Riera, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Source+Analysis+of+Interictal+Epileptiform+Discharges+Using+a+Rat+Model+of+Focal+Epilepsy.">Source Analysis of Interictal Epileptiform Discharges Using a Rat Model of Focal Epilepsy.</a> Organization for Human Brain Mapping. , (2014).</li><li>Birot, G., 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=Head+model+and+electrical+source+imaging:+A+study+of+38+epileptic+patients.">Head model and electrical source imaging: A study of 38 epileptic patients.</a> NeuroImage: Clinical. 16 (5), 77-83 (2014).</li><li>Riera, J. J., 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=Pitfalls+in+the+dipolar+model+for+the+neocortical+EEG+sources.">Pitfalls in the dipolar model for the neocortical EEG sources.</a> Journal of Neurophysiology. , (2012).</li><li>Hawrylycz, M., 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=The+Allen+Brain+Atlas.">The Allen Brain Atlas.</a> Springer Handbook of Bio-Neuroinformatics. , 1111-1126 (2014).</li><li>Schweinhardt, P., Fransson, P., Olson, L., Spenger, C., Andersson, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+template+for+spatial+normalization+of+MR+images+of+the+rat+brain.">A template for spatial normalization of MR images of the rat brain.</a> Journal of Neuroscience Methods. 129 (2), 105-113 (2003).</li><li>Schwarz, A. J., 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+stereotaxic+MRI+template+set+for+the+rat+brain+with+tissue+class+distribution+maps+and+co-registered+anatomical+atlas:+application+to+pharmacological+MRI.">A stereotaxic MRI template set for the rat brain with tissue class distribution maps and co-registered anatomical atlas: application to pharmacological MRI.</a> Neuroimage. 32 (2), 538-550 (2006).</li></ol>

没有利益冲突的声明。

一种新的方法，以非侵入性地记录多信道脑电图局灶性癫痫的特定临床前模型中进行说明。用于记录和分析程序的详情，与具体的实验提示，提供。有考虑取得成功结果的关键因素。首先，脑电图记录，获得高品质的信号是必不可少的。脑电图糊的适当粘度应该迷你帽制备过程中施加到每个电极，以及鼠的头部和耳朵发应在剃刮期间被完全除去。阻抗检查是为了确认脑电图记录质量的最重要的一步。其次，脑成像来源，产生量适当导体模式是至关重要的。每个表面应共登记。此外，所产生的电极的位置应具有对大鼠的头皮的实际电极的位置的最小距离误差。 尽管这个手稿源介绍采用头脑风暴12分析程序，他们可以使用其他软件开放16,17和商业产品18,19进行。此外，除了sLORETA 13，其他逆解决方案，如多偶极模型和波束形成器可用于4。 这种方法的一个限制是，行为分析，不能进行自脑电图记录镇静下进行。然而，相对于其它方法的脑电图记录大鼠5,6，这种方法是非侵入性的。 我们的初步结果支持为从脑电图记录IED标记，以确定在局灶性癫痫大鼠的刺激性区，以及评估其与癫痫发作起始11的基本机制关系的精确分类的重要性。此外，已经表明，脑电图源定位为这样的特定的IED表现出良好的对应关系与RESPective BOLD激活和去激活区20。 我们的研究将刺激使用临床前模型来评价生物医学工程师研制床工作台床的策略。例如，时下是手动的医院，这需要大量的人力精力进行IED提取。在这项研究中提出的方法自动完成。我们推测，当施加到患者FCD使用这种方法会产生类似的结果。我们正在准备的IRB协议的这个评价和对人的数据集的方法的其他方面。 此外，使用临床前模型将帮助我们理解在癫痫脑电图21源定位的能力和局限。脑源下属的癫痫准确的估计是治疗策略和手术规划至关重要。此外，具有大鼠脑电记录一个标准的平台将是有益的几种抗癫痫药物在临床前试验中的效力的评估。这是第一个研究中，癫痫的大鼠都记录非侵入镇静下，这将打开新的大门脑电图生物标志物用于癫痫的评价。然而，在该研究中提出的整个方法可延伸到其他的实验条件和脑部疾病。脑电图迷你帽，也可以使用在其他啮齿动物的类型。 在过去，在Wistar大鼠中一个前爪刺激范例已被用于评估记录的脑电图迷你盖2的数据的质量和再现性。此外，验证为脑源重建已执行从高分辨率颅骨脑电图同时录制与来自Wistar大鼠层局部场电位下一个晶须刺激范例22。这种方法已经开发Wistar大鼠，因为核磁共振图谱的存在对于这个特殊的鼠s火车。但是，它可以被应用到其它啮齿动物类型与图谱包括鼠标23，Sprague-Dawley大鼠24，以及Paxinos和Watson大鼠25的自己的标准格式。此外，我们所提出的方法的基本程序可在任何啮齿类动物临床前模型的量脑电​​图是一个重要的方式被使用。然而，这种方法的许多方面都特别为癫痫，特别是那些与脑电图预处理（IED检测和分类）。此外，研究人员必须知道在不同情况下使用镇静药物的正确。在我们的研究中使用异氟醚和dexdomitor已精心由于对简易爆炸装置的影响减少考虑。关于脑电图记录，在小鼠的情况下，相对小的头皮表面面积将大大减少信道数。

It has been shown that interictal epileptiform discharges (IEDs) observed from EEG constitute useful markers of epileptogenesis in patients with focal epilepsy3. The regions inside the brain from which these IEDs originate, named irritative zones, can in practice be localized based on EEG recordings4. Preclinical models are essential to find correlates between these irritative zones and the actual regions underlying seizure initiation. However, recording EEG from small animals is challenging because of the small surface area of the head compared to the human scalp. Although invasive methods for chronic recording in rats can be used5, 6, techniques are not available at this moment to acquire traditional EEG recordings on rodents under acute conditions without the need of anesthesia.
To solve this problem, we apply a patented EEG mini-cap2, which allows us to record 32-channel EEG data from rodents noninvasively. In this study, we also provide evidence about the need of an analgesic to preserve IED frequency. Therefore, although fixation of EEG mini-cap was performed under isoflurane, EEG recordings were obtained with rats only under sedation (dexdomitor)7. The method proposed in this study can be used in any preclinical rat model of focal epilepsy. To illustrate the capabilities of this methodology, we apply it to understand the correlates between irritative and seizure-onset zones in focal cortical dysplasia (FCD). To that end, we use a &#8220;double-hit&#8221; model of FCD8 in Wistar rats.
To perform brain source analysis, it is required to: a) accurately extract IEDs from EEG raw data and b) obtain a volume conductor model for the individual animal head. To generate a practical volume conductor model, we use an in vivo rat MRI atlas, comprising average images of intensity/shape and obtained via non-linear registration of T2 images of 31 Wistar rats9. The forward model for the generated volume conductor was computed by boundary element method (BEM)10. As in the case of humans, two typical patterns of IEDs (sharp-waves and spikes) were detected and sub-classified into different clusters through an intelligent feature extraction, feature selection, and classification process11. These sub-classified signals are used to estimate the brain source localizations associated with different types of irritative zones. We present the source analysis steps using a well-known public software called Brainstorm12. The EEG source localizations for each IED sub-type and the seizure onset time frames were performed using standardized low-resolution brain electromagnetic tomography (sLORETA)13, which is available in Brainstorm.

<table><tbody><tr><td>Data Acquisition Computer</td><td>Hewlett-Packard</td><td>Z210 Workstation</td><td></td></tr><tr><td>Dexdomitor</td><td>Orion Pharma</td><td>6295000</td><td>Dexmedetomidine hydrochloride</td></tr><tr><td>EEG Analysis Software</td><td>The Mathworks Inc.</td><td>MATLAB R2011b</td><td></td></tr><tr><td></td><td></td><td>Brainstorm</td><td>Sylvain et al. 2001</td></tr><tr><td></td><td></td><td>OpenMEEG</td><td>Gramfort et al. 2010</td></tr><tr><td>EEG Data Streamer</td><td>Tucker-Davis Technologies</td><td>RS4 Data Streamer</td><td></td></tr><tr><td>EEG Electrode Paste</td><td>Biotach</td><td>YGB 103</td><td></td></tr><tr><td>EEG Preamplifier</td><td>BioSemi</td><td>Active Two</td><td></td></tr><tr><td></td><td>Brain Products</td><td>BrainAmp</td><td></td></tr><tr><td></td><td>Tucker-Davis Technologies</td><td>PZ3 Low Impedance Amplifier</td><td></td></tr><tr><td>EEG Recording Software</td><td>BioSemi</td><td>ActiView</td><td></td></tr><tr><td>EEG Recording Software</td><td>Tucker-Davis Technologies</td><td>OpenEx - OpenDeveloper</td><td></td></tr><tr><td>EEG SCSI Connector</td><td>BioSemi</td><td>Active Two SCSI Connector</td><td></td></tr><tr><td></td><td>Brain Products</td><td>D-sub Connector</td><td></td></tr><tr><td>EEG Processor</td><td>Tucker-Davis Technologies</td><td>RZ2 BioAmp Processor</td><td></td></tr><tr><td></td><td>Tucker-Davis Technologies</td><td>Zif-Clif Digital Headstage</td><td></td></tr><tr><td>High Resolution EEG Mini-cap</td><td>Cortech Solutions</td><td>DA-AR-ELRCS32</td><td>US patent Application No. 13/641,834</td></tr><tr><td>Isoflurane, USP</td><td>VedcoPiramal Healthcare</td><td>NDC 66794-013-25</td><td></td></tr><tr><td>Isopropyl Alcohol</td><td>Aqua Solutions</td><td>3112213</td><td>90% v/v solution</td></tr><tr><td>Lubricant Ophthalmic Ointment</td><td>Rugby</td><td>NDC 0536-6550-91</td><td>Sterile</td></tr><tr><td>NaCl</td><td>Abbott</td><td>2B8203</td><td>Vaterinary 0.9% Sodium Chroride Injection USP</td></tr><tr><td>Physiology Recording Software</td><td>ADInstruments</td><td>LabChart 7.0</td><td></td></tr><tr><td>Physiology Recording System</td><td>ADInstruments</td><td>PowerLab 8/35</td><td></td></tr><tr><td>Syringe</td><td>Monoject</td><td>200555</td><td>12cc</td></tr></tbody></table>

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.

一旦所有的程序都适当地完成，估计的源可以被可视化的临床前模型的脑表面。 图5示出了从一个特定子类型尖峰（上）和从IED的尖波（底）的估计的来源。此外， 图6显示发作期间建立在连续的时间帧的源分布变化如何。这些结果支持所提出的方法的能力，以记录高分辨率脑电图对大鼠局灶性癫痫和使用记录的EEG进行来源分析。 <img alt="6视觉" src="/files/ftp_upload/52700/52700fig5.jpg" /> 的IED相对于不同簇中的尖峰（上）和尖波（底部）图5.估计脑源位置。（A）的时间序列，（B）的脑电图地形，和（C）的皮质电流酸CES。进行评价的结果，在标有一个红色垂直线（A）的一个特定的时间进行。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52700/52700fig5highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="6视觉" src="/files/ftp_upload/52700/52700fig6.jpg" /> 在检图6.大脑估算来源的时刻被标记为红色的垂直线。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/52700/52700fig6highres.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Brain Source Imaging in Preclinical Rat Models of Focal Epilepsy using High-Resolution EEG Recordings at JoVE.com

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

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 ...

brain-source-imaging-preclinical-rat-models-focal-epilepsy-using-high

Universidad San Sebastian

Research

JoVE Journal

Medicine

15.2K 次观看.  Florida International University. 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.

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

Simultaneous fMRI and Electrophysiology in the Rodent Brain

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

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

同时在啮齿动物脑功能磁共振成像和电

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

科研

神经科学

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 ...

Non-restraining EEG Radiotelemetry: Epidural and Deep Intracerebral Stereotaxic EEG Electrode Placement

Implantable EEG radiotelemetry is of central relevance in the neurological characterization of transgenic mouse models of neuropsychiatric and neurodegenerative diseases as well as epilepsies. This powerful technique does not only provide valuable insights into the underlying pathophysiological mechanisms, i.e., the etiopathogenesis of CNS related diseases, it also facilitates the development of new translational, i.e., therapeutic approaches. Whereas competing techniques that make use of recorder systems used in jackets or tethered systems suffer from their unphysiological restraining to semi-restraining character, radiotelemetric EEG recordings overcome these disadvantages. Technically, implantable EEG radiotelemetry allows for precise and highly sensitive measurement of epidural and deep, intracerebral EEGs under various physiological and pathophysiological conditions. First, we present a detailed protocol of a straight forward, successful, quick and efficient technique for epidural (surface) EEG recordings resulting in high-quality electrocorticograms. Second, we demonstrate how to implant deep, intracerebral EEG electrodes, e.g., in the hippocampus (electrohippocampogram). For both approaches, a computerized 3D stereotaxic electrode implantation system is used. The radiofrequency transmitter itself is implanted into a subcutaneous pouch in both mice and rats. Special attention also has to be paid to pre-, peri- and postoperative treatment of the experimental animals. Preoperative preparation of mice and rats, suitable anesthesia as well as postoperative treatment and pain management are described in detail.

Non-restraining EEG radiotelemetry is a valuable methodological approach to record in vivo long-term electroencephalograms from freely moving rodents. This detailed protocol describes stereotaxic epidural and deep intracerebral electrode placement in different brain regions in order to obtain reliable recordings of CNS rhythmicity and CNS related behavioral stages.

非约束脑电遥测:硬膜外和脑深部脑电立体定位放置电极

Implantable EEG radiotelemetry is of central relevance in the neurological characterization of transgenic mouse models of neuropsychiatric and ...

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

Advanced electroencephalographic analysis techniques requiring high spatial resolution, including electrical source imaging and measures of network connectivity, are applicable to an expanding variety of questions in neuroscience. Performing these kinds of analyses in a rodent model requires higher electrode density than traditional screw electrodes can accomplish. While higher-density electroencephalographic montages for rodents exist, they are of limited availability to most researchers, are not robust enough for repeated experiments over an extended period of time, or are limited to use in anesthetized rodents.1-3 A proposed low-cost method for constructing a durable, high-count, transcranial electrode array, consisting of bilaterally implantable headpieces is investigated as a means to perform advanced electroencephalogram analyses in mice or rats.

Procedures for headpiece fabrication and surgical implantation necessary to produce high signal to noise, low-impedance electroencephalographic and electromyographic signals are presented. While the methodology is useful in both rats and mice, this manuscript focuses on the more challenging implementation for the smaller mouse skull. Freely moving mice are only tethered to cables via a common adapter during recording. One version of this electrode system that includes 26 electroencephalographic channels and 4 electromyographic channels is described below.

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 ...

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 ...

Recording and Modulation of Epileptiform Activity in Rodent Brain Slices Coupled to Microelectrode Arrays

Temporal lobe epilepsy (TLE) is the most common partial complex epileptic syndrome and the least responsive to medications. Deep brain stimulation (DBS) is a promising approach when pharmacological treatment fails or neurosurgery is not recommended. Acute brain slices coupled to microelectrode arrays (MEAs) represent a valuable tool to study neuronal network interactions and their modulation by electrical stimulation. As compared to conventional extracellular recording techniques, they provide the added advantages of a greater number of observation points and a known inter-electrode distance, which allow studying the propagation path and speed of electrophysiological signals. However, tissue oxygenation may be greatly impaired during MEA recording, requiring a high perfusion rate, which comes at the cost of decreased signal-to-noise ratio and higher oscillations in the experimental temperature. Electrical stimulation further stresses the brain tissue, making it difficult to pursue prolonged recording/stimulation epochs. Moreover, electrical modulation of brain slice activity needs to target specific structures/pathways within the brain slice, requiring that electrode mapping be easily and quickly performed live during the experiment. Here, we illustrate how to perform the recording and electrical modulation of 4-aminopyridine (4AP)-induced epileptiform activity in rodent brain slices using planar MEAs. We show that the brain tissue obtained from mice outperforms rat brain tissue and is thus better suited for MEA experiments. This protocol guarantees the generation and maintenance of a stable epileptiform pattern that faithfully reproduces the electrophysiological features observed with conventional field potential recording, persists for several hours, and outlasts sustained electrical stimulation for prolonged epochs. Tissue viability throughout the experiment is achieved thanks to the use of a small-volume custom recording chamber allowing for laminar flow and quick solution exchange even at low (1 mL/min) perfusion rates. Quick MEA mapping for real-time monitoring and selection of stimulating electrodes is performed by a custom graphic user interface (GUI).

We illustrate how to perform recording and electrical modulation of 4-aminopyridine-induced epileptiform activity in rodent brain slices using microelectrode arrays. A custom recording chamber maintains tissue viability throughout prolonged experimental sessions. Live electrode mapping and selection of stimulating pairs are performed by a custom graphical user interface.

鼠脑切片偶联微电极阵列癫痫活性的记录与调控

Temporal lobe epilepsy (TLE) is the most common partial complex epileptic syndrome and the least responsive to medications. Deep brain stimulation (DBS) ...

Equipment Setup and Artifact Removal for Simultaneous Electroencephalogram and Functional Magnetic Resonance Imaging for Clinical Review in Epilepsy

Simultaneous electroencephalogram and functional magnetic resonance imaging (EEG-fMRI) is a unique combined technique that provides synergy in the understanding and localization of seizure onset in epilepsy. However, reported experimental protocols for EEG-fMRI recordings fail to address details about conducting such procedures on epilepsy patients. In addition, these protocols are limited solely to research settings. To fill the gap between patient monitoring in an epilepsy monitoring unit (EMU) and conducting research with an epilepsy patient, we introduce a unique EEG-fMRI recording protocol of epilepsy during the interictal period. The use of an MR conditional electrode set, which can also be used in the EMU for a simultaneous scalp EEG and video recording, allows an easy transition of EEG recordings from the EMU to the scanning room for concurrent EEG-fMRI recordings. Details on the recording procedures using this specific MR conditional electrode set are provided. In addition, the study explains step-by-step EEG processing procedures to remove the imaging artifacts, which can then be used for clinical review. This experimental protocol promotes an amendment to the conventional EEG-fMRI recording for enhanced applicability in both clinical (i.e., EMU) and research settings. Furthermore, this protocol provides the potential to expand this modality to postictal EEG-fMRI recordings in the clinical setting.

This article details simultaneous electroencephalogram and functional magnetic resonance imaging (EEG-fMRI) recording procedures that can be used in both clinical and research settings. EEG processing procedures to remove imaging artifacts for clinical review are also included. This study focuses on the example of epilepsy during the interictal period.

同步脑电图和功能性磁共振成像的设备设置和伪影去除，用于癫痫临床审查

Simultaneous electroencephalogram and functional magnetic resonance imaging (EEG-fMRI) is a unique combined technique that provides synergy in the ...

生物工程

<meta charset="utf8"></head><body>将 MEG 和 HD-EEG 整合到儿科癫痫定位中

Electromagnetic Source Imaging in Presurgical Evaluation of Children with Drug-Resistant Epilepsy

For children with drug-resistant epilepsy (DRE), seizure freedom relies on the delineation and resection (or ablation/disconnection) of the epileptogenic zone (EZ) while preserving the eloquent brain areas. The development of a reliable and noninvasive localization method that provides clinically useful information for the localization of the EZ is, therefore, crucial to achieving successful surgical outcomes. Electric and magnetic source imaging (ESI and MSI) have been increasingly utilized in the presurgical evaluation of these patients showing promising findings in the delineation of epileptogenic as well as eloquent brain areas. Moreover, the combination of ESI and MSI into a single solution, namely electromagnetic source imaging (EMSI), performed on simultaneous high-density electroencephalography (HD-EEG) and magnetoencephalography (MEG) recordings has shown higher source localization accuracy than either modality alone. Despite these encouraging findings, such techniques are performed in only a few tertiary epilepsy centers, are rarely recorded simultaneously, and are underutilized in pediatric cohorts. This study illustrates the experimental setup for recording simultaneous MEG and HD-EEG data as well as the methodological framework for analyzing these data aiming to localize the irritative zone, the seizure onset zone, and eloquent brain areas in children with DRE. More specifically, the experimental setups are presented for (i) recording and localizing interictal and ictal epileptiform activity during sleep and (ii) recording visual-, motor-, auditory-, and somatosensory-evoked responses and mapping relevant eloquent brain areas (i.e., visual, motor, auditory, and somatosensory) during visuomotor task, as well as auditory and somatosensory stimulations. Detailed steps of the data analysis pipeline are further presented for performing EMSI as well as individual ESI and MSI using equivalent current dipole (ECD) and dynamic statistical parametric mapping (dSPM).

<meta charset="utf8"></head><body>作者聚焦:通过新型生物标志物和增强定位推进儿童癫痫手术

Magnetoencephalography (MEG) and high-density electroencephalography (HD-EEG) are rarely recorded simultaneously, although they yield confirmatory and complementary information. Here, we illustrate the experimental setup for recording simultaneous MEG and HD-EEG and the methodology for analyzing these data aiming to localize epileptogenic and eloquent brain areas in children with drug-resistant epilepsy.

电磁源显像在儿童耐药性癫痫术前评估中的应用

For children with drug-resistant epilepsy (DRE), seizure freedom relies on the delineation and resection (or ablation/disconnection) of the epileptogenic ...

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

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啮齿动物联合局部脑电图和局部电位的同步记录

鼠脑切片偶联微电极阵列癫痫活性的记录与调控

同步脑电图和功能性磁共振成像的设备设置和伪影去除，用于癫痫临床审查

电磁源显像在儿童耐药性癫痫术前评估中的应用

电磁源显像在儿童耐药性癫痫术前评估中的应用

电磁源显像在儿童耐药性癫痫术前评估中的应用