We thank members of the Wilson lab for their helpful advice on the fabrication method.
This work was supported by the Simons Foundation, a NIH pathway to independence career award from the NINDS and a NARSAD Young Investigator Award (to M.M.H.) as well as grants from the NIH (to M.A.W.).

1，设计意图<ol><li>通过电子鼠脑图谱的矢状切面滚动标识选择（外侧膝状体（LGN，视觉丘脑））的大脑区域。 </li><li>在/ P的坐标（-2.3 - -2.7毫米）时，LGN是最宽的。使用此区域来设计驱动器底部（底部件）。 注:共有8独立地可动电极可用于靶向LGN（4-6电极将使它的LGN，2-4电极被加到偏移注入误差， 图1A）。 </li><li>在SolidWorks中，在前面的平面绘制设计体（ 图1B）的草图。单击草图，然后用直线和曲线相结合，绘制草图，其中将包括等高线的驱动器底座，把手和聚酰胺半槽，如图所示。确保该轮廓不包含任何开口间隙。然后点击退出草图。 </li><li>接下来，选择正面和右侧面，然后单击"克雷亚TE轴"。然后，通过转动突出蓝色草图轮廓（ 图1B）360°创建的三维设计的人体模型。在功能菜单中，单击"旋转凸台/基体"。选择中线为旋转轴。在参数部分，下方向1单击失明，在选择角度360.00度。在所选择的轮廓部分，确保蓝色高亮轮廓是选择之一。 </li><li>通过旋转红色创建一个聚酰亚胺半插槽突出轮廓13°（ 图1C，左上图）。步骤是相同的​​，以1.4以上除角规格</li><li>创建一个驱动器处理由旋转的绿色轮廓15°（ 图1C，右上图）。 </li><li>创建使用循环模式功能（ 图1C，左下）第二个驱动器手柄。在功能菜单中，单击"循环模式"。在参数中，选择中线为革命轴。选择180.00度的角度，和2作为实例的数量。保证第一柄下选择"功能，以模式"。 </li><li>创建16聚酰亚胺使用循环模式功能（ 图1C，左下）半插槽。执行类似的动作，以1.7的，但选择的第一个聚酰亚胺半槽作为"功能，模式"。的夹角为22.5°和实例的数量是16（注意:这仅仅是360°通过的次数除以要图案的特征） </li><li>建立在其上绘制的聚酰亚胺插座一个新的平面。通过点击主菜单上的"插入"实现这一目标。单击"参考几何体"，选择聚酰亚胺半槽两侧，然后单击"创建新平面"; （ 图1D，上图） </li><li>创建微硬盘插座（螺丝孔，聚酰亚胺孔和抗扭矩轨（ 图1D，下图）。通过创建实现这一草图，它包括在1.9中创建的新飞机所有这些功能。注意，对于抗扭矩导轨，在聚酰胺外层狭缝的两侧之间限定的中心线。然后，通过创建两个圆在垂直于中心线，其中心是1半径分开，然后修整中间轮廓绘制的抗扭矩导轨。 </li><li>在功能菜单中，单击"拉伸凸台/基"创建反扭矩轨道，并选择10毫米走上坡路和2mm向下去盲目拉伸。对于螺纹孔和聚酰亚胺孔，点击"拉伸剪切"，然后选择盲目6毫米，和几个MMS走上坡路为（ 图1E，左）。 </li><li>图案的微型驱动器容器16倍，使用该中心作为旋转（22.5°，16个实例，间距相等）的轴线（ 图1E，右） </li><li>在手柄的上方，画3毫米x 3毫米中开始驱动器手柄的中央尖，面向中部的一个红双喜。挤出这个2毫米以上使用"拉伸老板"的功能。绘制直径为1mm圈的位置，其中欧洲投资银行螺丝将继续下去。随后，让1.5毫米"拉伸切除"，使一个洞。然后，图案采用圆形图案的功能（文本叠加:180°，2个实例，间距相等，围绕中心轴线）的两倍框和孔。 </li><li>使用图1F的尺寸（单位:毫米）画一个顶件草图。使用"拉伸凸台/基"，以它的3D模型。 注意:在这些步骤中的驱动器设计完成。的物理驱动体通过立体平版印刷的过程中创建。有许多公司提供基于STL文件立体印刷。我们建议，所用硬质塑料打印（如Accura®分55），具有至少为0.1毫米的最小解析服务。 </li></ol> 2，准备工作超空间引擎组件<ol><li>奠定了一个小（:&#39;/ 0116&#39;内径/外径0.0071&#39;;墙:0.00225）一块在一个平面上双面胶带，切为31g聚酰亚胺管所需的数量约8厘米（ 图2A - 2B） 。 </li><li>铺陈导引管的​​第一层上的双面胶带，小心地放置导管尽可能接近彼此在磁带上。轻拍过的聚酰亚胺层有少量的薄，氰基丙烯酸酯胶水。 （ 图2C） </li><li>迅速奠定了聚酰亚胺的第二层（ 图2D）。 </li><li>创建使用＆G套管光纤预留位置。确保这是使用前的集会（ 图2E）被纳入聚四氟乙烯基润滑剂润滑。 </li><li>适用一行环氧树脂4-5毫米长垂直于聚酰亚胺包（ 图2F）。一旦环氧树脂硬化（2-3小时），从底层和稀土元素除去胶带丙氧基的另一面。后的环氧树脂再次硬化，在＆G插管可以被删除，用剃刀刀片（ 图2G），得到两种聚酰亚胺矩阵，其中每一个可被用于一个超空间（ 图2H）的构建体切在中间。 </li><li>打印出来的透明纸上锥模板和削减重型铝箔（ 图3A - 3C）对应的表。 </li><li>涂上一层环氧树脂，以铝箔和快速应用透明纸。用重物或木销钉，理顺环氧树脂，使其均匀地分布（ 图3D）。 </li><li>切出圆锥形的模板，并夹在一起用鳄鱼夹。最后，使用环氧树脂的另一民建联永久固定件（ 图3E）。 </li></ol>该微型硬盘的3总装<ol><li>连接欧洲投资银行向驱动体，并重新插入，通过聚酰亚胺导管矩阵＆G套管。配合使用的EIB的光纤孔，以确保引导管是垂直于EIB驱动体的聚酰亚胺基体和环氧树脂基体与驱动体小心，以确保没有环氧树脂流入导向管或进驱动体（ 图4A - 4C）。 </li><li>在聚酰亚胺基体的每个导​​向管映射到驱动器主体的内壁上的相应的支架上。滑动的33克的聚酰亚胺小环在每个导管，进入支架和涂抹少量的氰基丙烯酸酯胶贴上每个导管。 （ 图4D - 4E）最后，环氧整个装置的驱动体的内壁和切聚酰亚胺，使它们伸出的正上方的内唇缘（ 图4F - 4G）。 </li><li>通过将自定义B中的一个构建微硬盘组装通过顶片后跟5毫米弹簧中的一个的中心孔uilt螺钉。滑动顶部件在轨道中的一个的外孔，缓缓地将螺丝。驱动器的螺钉，直到弹簧达到它的最小压缩长度。 （ 图4H - 4I）重复此过程，每个轨道/ Microdrive微型硬盘（ 图4J）。 </li><li>打开驱动器阵列倒过来，把导管矩阵的图片。此图像将被用于映射对应于各微驱动器（ 图4K）的导向管的位置。 </li><li>插入的聚酰亚胺管（0.005"），在每个导向管从驱动器基座的底部。让运营商管从完全降下微硬盘，并记录在照片上相应的微驱动器的身份顶端一直延伸1-2毫米。 （ 图4L - 4M） </li><li>环氧聚酰亚胺管的微硬盘支持，注意不要勒吨环氧树脂通过微硬盘上的弹簧或螺丝（ - 4Q图4N，4P）运行。 </li><li>全面降低所有的微型硬盘。切断所有的聚酰亚胺管关闭冲洗在聚酰亚胺基体（ 图40）的底部。 </li><li>安装电接口板使用两个＃00-90点¯x3月16日&#39;螺丝钉（ 图4R）驱动器基地。 注意:此时驱动器阵列已准备好装有stereotrodes或四极管。关于四极结构和加载的详细信息，请参阅10。打印驱动基地和微型硬盘被设计在SolidWorks 2011 3D CAD软件:下载链接SolidWorks文件。 </li><li>装载后，反转驱动和小心降低屏蔽锥体过驱动，使得只有底部片伸出。由epoxying锥到驱动体粘贴遮蔽锥体。 </li><li>经过锥体连接，脱衣小长不锈钢丝（.008"&#39;裸，0.011&#39;涂层）和引脚，以欧洲投资银行。划伤用针的锥体内，铝部分和使用银粉漆钢丝研磨成锥形。一旦银粉漆干燥后，加强与环氧树脂的民建联。或者，该钢丝可以被直接附着在锥体与导电环氧树脂的DAB（MG化学品，萨里，加拿大）。 </li></ol>

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作者什么都没有透露。

这个协议概述构建一个超轻质微驱动器阵列，用于针对一个或多个脑区域中的鼠标的过程。后的最终步骤的建设，超空间是准备使用标准外科植入技术植入和固定在小鼠的颅骨与牙科用粘固剂。邮政植入，电极可以各自独立地推进用小螺丝刀，而鼠标的手约束。每个电极的进步是由螺杆的螺距确定的每匝之间的距离。使用此处所引用的螺钉进入每个电极每圈大约150毫米，虽然半和四分之一圈可以用于更大的分辨率。 在图1B中的草图的尺寸确定所述植入物的整体尺寸，因此，一个显而易见的方法进行双向缩放植入物是改变在该临界草图尺寸。此外，日螺钉Ë长度可以延伸到目标更深的大脑结构。我们建议您定制的钛螺丝，那些轻便，比钢更不易碎。注意，反扭矩导轨需要与螺杆长度线性比例，并且在这一点上，我们还没有确定的最大长度在这些结构可以被打印。到目标的多个脑区域中，底部件的形状可以被修改。在加入已知大小的垫圈（厚度200μm）时，可以提供针对单独的大脑结构（例如，海马和前额叶皮质）的聚酰亚胺之间所需间隔物。这些可以包括在底部件的组装步骤，并且以后切断环氧树脂硬化后。 这种设计的很大的局限性是其上​​的专用软件（在这种情况下Solidworks的）的依赖。提供用户友好的界面有利于以最小的工程backg设计这种设备的开放源码项目的未来发展一轮将是巨大的利益在神经科学界。 此方法提供了优于现有方法的几个优点。首先，设计是简单的，依赖于极少数的草图（ 图1）。第二，它是超轻，不需要牙科用粘固剂或重材料进入其组件。总体而言，它重约1.7克 - 类似功能的市售植入物重量的近三分之一。第三，它不需要专门的设备，以使 - 植入物本体可以三维打印来自多个源（例如approto.com，但也有几个其他的）;螺丝可定制（例如antrinonline.com）;弹簧是可商购的（例如leesprings.com）;其结果是，整个组装过程可以在一天内发生。最后，这些植入物已被用于从天然觅食过程中的多个脑区的记录，结构化行为的任务和睡眠（ 图5）。 这种方法的未来应用包括实施其扩展性。它是可能的植入物可以双向通过简单地改变1缩放的）草图图1B和图案化的微驱动器容座（ 图1D）的，2）的数量的大小。例如，它可以向下缩放以记录从自由活动小鼠早在发展，并且缩放向上从大鼠，兔，雪貂，也许非人灵长类动物来记录。 最后一个字是​​提醒读者，要成功实施概括的方法是至关重要的原型，他们落实.STL附加设计文件的任何修改。读者将注意到，例如，该附设计包含了"数字8"的反扭矩轨。这是最好的设计可能会带来3D打印的限制，因为它往往要求我们钻这些漏洞。有它是一个圆，将COMPromise稳定，但有它是一个正方形或有角度的形状会限制来解决3D打印缺陷钻井的能力。

小家鼠有，由于其遗传易处理，迅速 ​​成为首选的动物模型有兴趣确定的基因神经元的微电路级的解剖和研究人类疾病的小鼠模型生理学家。例如，近期出台的致病性遗传工具，如光遗传学，化学遗传学驱动器允许实验者测试确定的神经回路的必要性和充分性的行为1-4。重组转基因小鼠驱动线（Cre的线），的广泛应用已扩增实验放心由神经元亚型有针对性，增加了鼠标的这些实验5的值。 同样地，遗传筛选和常见的神经障碍和精神障碍的基因组范围关联提供了便利的遗传风险因素的识别脑疾病6,7。这些技术进步，再加上生长工具箱的遗传操作和小鼠基因组工程，都取得了它选择的生物体为模拟人类疾病。疾病模型和致病性遗传工具的组合，为了解脑部疾病和识别电路级目标的干预了前所未有的机遇。 为了充分利用这些分子的工具，并深入了解健康与疾病微电路的功能，就必须对他们夫妇与大脑活动的生理读数。理想情况下，实验者将能够监视大量的神经元，同时保持单个细胞的分辨率。在自由活动的动物外，多电极记录提供了这样的机会;然而，在小鼠中使用这种技术受到了限制。从小型目标（ 例如 ，在海马CA1区层）记录，使用可调式电极是必要的，下面surgic小变动记录电极人注入使它不可能保持记录稳定性8,9。传统上，已经使用的移动至大脑内的电极施加重量的限制，在小鼠中使用时的方法，使得它具有挑战性的大量神经元在这个生物体行为的几个记录。 在这里，方法进行了介绍用于制造小型，超轻型，微电极阵列是单独定制的大脑区域被瞄准，光遗传学兼容，并从市售品容易组装。内的多电极"超空间"每个"微驱动器"利用一个弹簧和螺旋机构以推进电极和一个塑料导轨，内置于超空间体，以从螺杆抵消扭矩。首先，设计的超空间机构和微型硬盘为3D打印的CAD程序的过程描述。通过设计定制超空间机构对于具体的结构，所以能够提高定位的精度，并进一步提高该制剂的产率。第二，在制造过程中进行了详细的方法，其中所述多电极阵列从零件是可商购的组装用手说明。这种技术已被使用，成功地从海马，丘脑和皮层中的自由活动动物神经元的天然觅食和操作性任务期间合奏记录。

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			<td height="22" style="height:22px;width:169px;">Microdrives</td>
			<td style="width:131px;"></td>
			<td style="width:143px;"></td>
			<td style="width:176px;">See .STL file</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:169px;">Drive Body</td>
			<td style="width:131px;"></td>
			<td style="width:143px;"></td>
			<td style="width:176px;">See .STL file</td>
		</tr>
		<tr height="20">
			<td height="81" rowspan="4" style="height:81px;width:169px;">Outer Polyimide Guide Tube</td>
			<td rowspan="4" style="width:131px;">Minvasive Components&#160;&#160;</td>
			<td rowspan="4" style="width:143px;">IWG Item # 72113300022-012</td>
			<td style="width:176px;">Length:12&#8217;&#8217;,&#160;</td>
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			<td height="20" style="height:20px;width:176px;">ID:.0071&#8217;&#8217;,&#160;</td>
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		<tr height="20">
			<td height="20" style="height:20px;width:176px;">OD:.0116&#8217;&#8217;,&#160;</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:176px;">WALL:.00225&#8217;&#8217;</td>
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		<tr height="20">
			<td height="81" rowspan="4" style="height:81px;width:169px;">Inner Polyimide Guide Tube</td>
			<td rowspan="4" style="width:131px;">Minvasive Components&#160;</td>
			<td rowspan="4" style="width:143px;">IWG Item # 72113900001-012</td>
			<td style="width:176px;">Length: 12&#8217;&#8217;,&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:176px;">ID:.0035&#8217;&#8217;,&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:176px;">OD:.0055&#8217;&#8217;,&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:176px;">WALL:.001&#8217;&#8217;</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:169px;">Grounding Wire</td>
			<td style="width:131px;">A-M Systems, Inc.</td>
			<td style="width:143px;">&#160;Catalog # 791900</td>
			<td style="width:176px;">.008&#39;&#39; Bare, .011&#39;&#39; Coated</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:169px;"></td>
			<td style="width:131px;"></td>
			<td style="width:143px;"></td>
			<td style="width:176px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:169px;">Tri-Flow&#160;</td>
			<td style="width:131px;"></td>
			<td style="width:143px;"></td>
			<td style="width:176px;">Teflon based lubricant - Aerosol</td>
		</tr>
		<tr height="40">
			<td height="181" rowspan="5" style="height:181px;width:169px;">Microdrive Springs</td>
			<td rowspan="5" style="width:131px;">Lee Spring</td>
			<td rowspan="5" style="width:143px;">Part # CB0050B 07 E</td>
			<td style="width:176px;">Outside Diameter: 1.016 mm</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:176px;">Hole Diameter: 1.193 mm</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:176px;">Wire Diameter: 0.127 mm</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:176px;">Free Length 10.160 mm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:176px;">Solid Length 3.581 mm</td>
		</tr>
		<tr height="59">
			<td height="80" rowspan="2" style="height:80px;width:169px;"><a name="RANGE!A24">Z-poxy 5 Minute</a></td>
			<td rowspan="2" style="width:131px;">Pacer Technology (Zap)</td>
			<td rowspan="2" style="width:143px;">PT37</td>
			<td rowspan="2" style="width:176px;"></td>
		</tr>
		<tr height="21">
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:169px;">&#160;Silver Paint</td>
			<td style="width:131px;">&#160;GC Electronics</td>
			<td style="width:143px;">&#160;Part #: 22-023</td>
			<td style="width:176px;">Silver Print II</td>
		</tr>
		<tr height="20">
			<td height="41" rowspan="2" style="height:41px;width:169px;"></td>
			<td rowspan="2" style="width:131px;">Tri-Flow&#160;</td>
			<td align="right" rowspan="2" style="width:143px;">20009</td>
			<td rowspan="2" style="width:176px;"></td>
		</tr>
		<tr height="21">
		</tr>
		<tr height="59">
			<td height="100" rowspan="3" style="height:100px;width:169px;">26 Gauge Hypodermic Tube - Stainless Steel</td>
			<td rowspan="3" style="width:131px;">Small Parts&#160;</td>
			<td rowspan="3" style="width:143px;">HTXX-26T-12-10</td>
			<td style="width:176px;">Length: 12&#8217;&#8217;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:176px;">ID: .012&#8217;&#8217;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:176px;">OD: .018&#8217;&#8217;</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:169px;">EIB screws</td>
			<td style="width:131px;">Component Supply Co.</td>
			<td style="width:143px;">MX-0090-03SP</td>
			<td style="width:176px;">#00-90 x 3/16&#8217;&#8217;</td>
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		<tr height="41">
			<td height="41" style="height:41px;width:169px;">Fine Scissors - Toughcut</td>
			<td style="width:131px;">Fine Science Tools</td>
			<td style="width:143px;">14058-09</td>
			<td style="width:176px;">22mm</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:169px;">Transparency Paper</td>
			<td style="width:131px;">3M</td>
			<td style="width:143px;">PP2500</td>
			<td style="width:176px;"></td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:169px;">Aluminum Foil</td>
			<td style="width:131px;">Reynold&#39;s Wrap Heavy Duty</td>
			<td style="width:143px;"></td>
			<td style="width:176px;">Extra Thick</td>
		</tr>
	</tbody>
</table>

design and fabrication of ultralight weight, adjustable multi-electrode probes for electrophysiological recordings in mice

生理研究中的老鼠， 家鼠的数量，在经历了近期的飙升，平行的基因定位于微电路解剖和疾病建模方法的发展。光遗传学的引入，例如，允许对识别的基因的神经元的双向操作，以前所未有的时间分辨率。为了充分利用这些工具，深入了解大脑微电路之间的动态交互，很重要的是一个人从神经元的合奏深深的这个小啮齿动物的大脑内记录，在这两个头固定和自由活动的准备能力。从深层结构和不同的细胞层记录需要准备，允许对期望的大脑区域的电极精确的进步。来记录神经套装，有必要使各电极是独立地移动，从而允许实验者以解决单个细胞同时留下neighbO形圈电极不受干扰。在自由表现的鼠标做到既需要一个电极驱动的轻便，有弹性，并针对特定的大脑结构高度可定制的。 的技术设计和制造的微型，超轻的重量，是单独定制的，从市售零件组装容易Microdrive微型电极阵列呈现。这些设备易于扩展和可定制的结构作为目标;它已被成功地用于从丘脑和皮层区域的过程中自然的行为自由的动物行为记录。

植入物构造是一个过程，与3D印刷超空间（ 图1）的设计开始，进行到所述底部件的结构（ 如图2所示 ），屏蔽锥体（ 图3），并在超空间中的最终组件中，由该微型硬盘的单项在建工程（ 图4）。其次是通过装载微型硬盘与电极（请参见10）这些步骤。以下这个步骤，就可以使用这些设备从多个脑区域来记录。在图5中，例如从同步录音外侧膝状体（LGN）和海马（HPC）的跟踪显示。 图5B所示的单单元的稳定性一直是显着的，显示在数天为一疗程一致的波形。这些神经元被确定为通过响应于发光二极管的刺激被LGN神经元，如由peristimulus时间直方图（PSTH）在图5C中 。在同一制剂，高性能计算局部场电位记录，作为国家行为的代理。这些迹线显示尖波涟漪（ 图5D），在此期间的行为静止，其海马原点一致。 <img alt="图1" src="/files/ftp_upload/51675/51675fig1highres.jpg" /> 图1设计在SolidWorks中的超空间引擎。小鼠大脑的A / P的冠状断面的A原理图坐标-2.3 - -2.7毫米前囟。四个人聚酰亚胺（300微米）的皮质上面绘制，说明LGN区域（红色）与电极的目标。设计机身B.素描。围绕着蓝色的轮廓180度导致了3D设计的人体模型（小图）。℃。添加聚酰亚胺插槽和驱动器的手柄设计机构。围绕着红极力推B中ighted轮廓由13°导致聚酰亚胺半插槽（左上）。一个驱动器手柄是由15°（右上）围绕着绿色的轮廓B中加入。第二把手，通过使用圆形图案功能（左下）中加入。相同的功能，可用于创建16聚酰亚胺半时隙（右下图）。D.一个新的平面将被添加到设计（顶部），从而创建了微驱动容器的新的草图，包括螺钉孔的，聚酰亚胺孔和反扭矩导轨（底部）。E.将这些功能使用剪切和拉伸功能实现到设计和旋转360°，以创建16个容器。顶部件草图（左）和3D模型的F.尺寸（右）。 <a href="https://www.jove.com/files/ftp_upload/51675/51675fig1large.jpg" target="_blank">请点击这里查看该图的放大版本。</a> <img alt="图2"SRC ="/文件/ ftp_upload / 51675 / 51675fig2highres.jpg"/&gt; 图2:准备底部件的超空间中。A.第一聚酰亚胺管放置到双面胶带。B.后续管被单独放置，注意尽量减少管之间的空间。C.之后，第一层被布置，一层薄薄的氰基丙烯酸酯胶应用于四聚酰亚胺的迅速增加胶水干燥之前的第二层。聚酰亚胺束E.在楼顶，＆G套管添加为占位光纤。˚F ，整个构建体被牢固地固定有一滴环氧树脂。G.切除插管后，将构建物可以在用剃刀刀片的中间切开，得到两个相同的底部件。H.视图上的成品的切割表面底部件，说明四聚酰亚胺的两个双行与空穴的光纤。 <a href="https://www.jove.com/files/ftp_upload/51675/51675fig2large.jpg" target="_blank">请点击这里查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/51675/51675fig3highres.jpg" /> 图3组装超空间。A.聚酰亚胺基体是使用＆G插管插入驱动器主体，并与所述电子接口板（EIB）对齐。B.一个少量的环氧树脂被用于贴上聚酰亚胺基质，以的驱动机构。C. A环氧的第二应用可能是必要的，在这之后过量的环氧应dremeled远D.顶视图上的驱动体与插入基体中。E.使用一小块33克的聚酰亚胺管，所述外导管连接在驱动机构的相应插槽。 楼 </st荣&gt;所有外导管应被映射到一个轨道，同时注意尽量减少张力管。G.毕竟在外导管映射，它们应该受到保护与环氧树脂和削减只是内唇上面。H. Microdrive卡组合件，包括一个定制的螺钉，一个5毫米的弹簧和顶片应装配并放置在对应于所述导向管中的一个的轨道。 一，每个微驱动组件应该仔细拧入驱动机构。J.组装完成后，每个导向管应该有一个相应的微硬盘聚酰亚胺基体的L K.底视图- 。 米聚酰亚胺管（0.005"）被插入到每个外引导管。N.每个内导管应该紧贴到叉的它的对应的微驱动。O的内聚酰亚胺管连接用环氧树脂为相应的微驱动器和切割尽可能短。毕竟内导管是用环氧树脂，内导管从聚酰亚胺基体应突出内导管装载过程中切齐与基体的嘴唇。P.倒宏观驱动的。 问:顶宏观驱动器期间内导管负载R。完全组装的超空间引擎与欧洲投资银行连接，随时可以装入电极。 <a href="https://www.jove.com/files/ftp_upload/51675/51675fig3large.jpg" target="_blank">请点击这里查看该图的放大版本。</a> <img alt="图4" src="/files/ftp_upload/51675/51675fig4highres.jpg" /> 图4，准备屏蔽锥锥A.模板打印在透明纸 B - D。铝箔的片材被粘合到用一薄层的环氧树脂的模板。 大肠杆菌&lt;<html> /强&gt;切出的模板后，将锥形成并用环氧树脂胶粘在一起。 <img alt="图5" src="/files/ftp_upload/51675/51675fig5highres.jpg" /> 图采用超轻量超空间引擎，一个5多点记录 。自由鼠表现与超空间引擎植入。两个单单元的二例图像波形记录从这个鼠标。C.左，小鼠大脑突出外侧膝状体，其中一些电极被下调的冠状切面。右，例如peristimulus 2 LGN神经元对齐视觉刺激（黄色条）的时间直方图（PSTHs）。D.右，冠状部突出海马（HPC），其中另一组电极被降低。右，局部场电位海马纹波（红色光）的记录实例。 NT"FO:保together.within页="总是"&gt; <img alt="图6" src="/files/ftp_upload/51675/51675fig6highres.jpg" /> 图6概述驱动组件（左）超空间分量的综合概观。 （右）插图NA个人微硬盘组装。

Watch this Scientific Journal Video about Design and Fabrication of Ultralight Weight, Adjustable Multi-electrode Probes for Electrophysiological Recordings in Mice at JoVE.com

Design and Fabrication of Ultralight Weight, Adjustable Multi-electrode Probes for Electrophysiological Recordings in Mice

Understanding the neural substrates of behavior requires brain circuit ensemble recording. Because of its genetic tractability, the mouse offers a model for circuit dissection and disease mimicry. Here, a method of designing and fabricating miniaturized probes is described that is suitable for targeting deep brain structure in the mouse.

设计和超轻重量的制作，可调节的多电极电生理记录小鼠探头

The number of physiological investigations in the mouse, mus musculus, has experienced a recent surge, paralleling the growth in methods of genetic ...

design-fabrication-ultralight-weight-adjustable-multi-electrode

Research

JoVE Journal

Neuroscience

17.6K 次观看.  New York University Langone Medical Center. Understanding the neural substrates of behavior requires brain circuit ensemble recording. Because of its genetic tractability, the mouse offers a model for circuit dissection and disease mimicry. Here, a method of designing and fabricating miniaturized probes is described that is suitable for targeting deep brain structure in the mouse.

视频: Design and Fabrication of Ultralight Weight, Adjustable Multi-electrode Probes for Electrophysiological Recordings in Mice

Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster

When startled adult D. melanogaster react by jumping into the air and flying away. In many invertebrate species, including D. melanogaster, the "escape" (or "startle") response during the adult stage is mediated by the multi-component neuronal circuit called the Giant Fiber System (GFS). The comparative large size of the neurons, their distinctive morphology and simple connectivity make the GFS an attractive model system for studying neuronal circuitry. The GFS pathway is composed of two bilaterally symmetrical Giant Fiber (GF) interneurons whose axons descend from the brain along the midline into the thoracic ganglion via the cervical connective. In the mesothoracic neuromere (T2) of the ventral ganglia the GFs form electro-chemical synapses with 1) the large medial dendrite of the ipsilateral motorneuron (TTMn) which drives the tergotrochanteral muscle (TTM), the main extensor for the mesothoracic femur/leg, and 2) the contralateral peripherally synapsing interneuron (PSI) which in turn forms chemical (cholinergic) synapses with the motorneurons (DLMns) of the dorsal longitudinal muscles (DLMs), the wing depressors. The neuronal pathway(s) to the dorsovental muscles (DVMs), the wing elevators, has not yet been worked out (the DLMs and DVMs are known jointly as indirect flight muscles - they are not attached directly to the wings, but rather move the wings indirectly by distorting the nearby thoracic cuticle) (King and Wyman, 1980; Allen et al., 2006). The di-synaptic activation of the DLMs (via PSI) causes a small but important delay in the timing of the contraction of these muscles relative to the monosynaptic activation of TTM (~0.5 ms) allowing the TTMs to first extend the femur and propel the fly off the ground. The TTMs simultaneously stretch-activate the DLMs which in turn mutually stretch-activate the DVMs for the duration of the flight. The GF pathway can be activated either indirectly by applying a sensory (e.g."air-puff" or "lights-off") stimulus, or directly by a supra-threshold electrical stimulus to the brain (described here). In both cases, an action potential reaches the TTMs and DLMs solely via the GFs, PSIs, and TTM/DLM motoneurons, although the TTMns and DLMns do have other, as yet unidentified, sensory inputs. Measuring "latency response" (the time between the stimulation and muscle depolarization) and the "following to high frequency stimulation" (the number of successful responses to a certain number of high frequency stimuli) provides a way to reproducibly and quantitatively assess the functional status of the GFS components, including both central synapses (GF-TTMn, GF-PSI, PSI-DLMn) and the chemical (glutamatergic) neuromuscular junctions (TTMn-TTM and DLMn-DLM). It has been used to identify genes involved in central synapse formation and to assess CNS function.

Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster

The Giant Fiber System is a simple neuronal circuit of adult Drosophila melanogaster containing the largest neurons in the fly. We describe the protocol for monitoring synaptic transmission through this pathway by recording post synaptic potentials in dorsal longitudinal (DLM) and tergotrochanteral (TTM) muscles following direct stimulation of the Giant Fiber interneurons.

从巨纤维通路的电生理记录 D。果蝇</em

When startled adult D. melanogaster react by jumping into the air and flying away. In many invertebrate species, including D. melanogaster, the "escape" ...

科研

神经科学

Multi-electrode Array Recordings of Neuronal Avalanches in Organotypic Cultures

The cortex is spontaneously active, even in the absence of any particular input or motor output. During development, this activity is important for the migration and differentiation of cortex cell types and the formation of neuronal connections1. In the mature animal, ongoing activity reflects the past and the present state of an animal into which sensory stimuli are seamlessly integrated to compute future actions. Thus, a clear understanding of the organization of ongoing i.e. spontaneous activity is a prerequisite to understand cortex function. 
Numerous recording techniques revealed that ongoing activity in cortex is comprised of many neurons whose individual activities transiently sum to larger events that can be detected in the local field potential (LFP) with extracellular microelectrodes, or in the electroencephalogram (EEG), the magnetoencephalogram (MEG), and the BOLD signal from functional magnetic resonance imaging (fMRI). The LFP is currently the method of choice when studying neuronal population activity with high temporal and spatial resolution at the mesoscopic scale (several thousands of neurons). At the extracellular microelectrode, locally synchronized activities of spatially neighbored neurons result in rapid deflections in the LFP up to several hundreds of microvolts. When using an array of microelectrodes, the organizations of such deflections can be conveniently monitored in space and time. 
Neuronal avalanches describe the scale-invariant spatiotemporal organization of ongoing neuronal activity in the brain2,3. They are specific to the superficial layers of cortex as established in vitro4,5, in vivo in the anesthetized rat 6, and in the awake monkey7. Importantly, both theoretical and empirical studies2,8-10 suggest that neuronal avalanches indicate an exquisitely balanced critical state dynamics of cortex that optimizes information transfer and information processing.
In order to study the mechanisms of neuronal avalanche development, maintenance, and regulation, in vitro preparations are highly beneficial, as they allow for stable recordings of avalanche activity under precisely controlled conditions. The current protocol describes how to study neuronal avalanches in vitro by taking advantage of superficial layer development in organotypic cortex cultures, i.e. slice cultures, grown on planar, integrated microelectrode arrays (MEA; see also 11-14).

A robust way to study neuronal avalanches, i.e. scale-invariant spatio-temporal activity bursts, indicative of critical state dynamics in cortex. Avalanches emerge spontaneously in developing superficial layers of cultured cortex which allows for long-term measurements of the activity with planar integrated multi-electrode arrays (MEA) under precisely controlled conditions.

器官型培养的神经元雪崩多电极阵列记录

The cortex is spontaneously active, even in the absence of any particular input or motor output.  During development, this activity is important for the ...

Design and Assembly of an Ultra-light Motorized Microdrive for Chronic Neural Recordings in Small Animals

The ability to chronically record from populations of neurons in freely behaving animals has proven an invaluable tool for dissecting the function of neural circuits underlying a variety of natural behaviors, including navigation1 , decision making 2,3, and the generation of complex motor sequences4,5,6. Advances in precision machining has allowed for the fabrication of light-weight devices suitable for chronic recordings in small animals, such as mice and songbirds. The ability to adjust the electrode position with small remotely controlled motors has further increased the recording yield in various behavioral contexts by reducing animal handling.6,7
 Here we describe a protocol to build an ultra-light motorized microdrive for long-term chronic recordings in small animals. Our design evolved from an earlier published version7, and has been adapted for ease-of use and cost-effectiveness to be more practical and accessible to a wide array of researchers. This proven design 8,9,10,11 allows for fine, remote positioning of electrodes over a range of ~ 5 mm and weighs less than 750 mg when fully assembled. We present the complete protocol for how to build and assemble these drives, including 3D CAD drawings for all custom microdrive components.

The design, fabrication and assembly of an ultra-light motorized microdrive is described. The device provides a cost-effective and easy-to-use solution for chronic recordings of single units in small behaving animals.

设计和组装的超轻型电动微型硬盘（Microdrive）慢性神经录音的小动物

The ability to chronically record from populations of neurons in freely behaving animals has proven an invaluable tool for dissecting the function of ...

Dual Electrophysiological Recordings of Synaptically-evoked Astroglial and Neuronal Responses in Acute Hippocampal Slices

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs through activation of their ion channels and neurotransmitter receptors, and process information in part through activity-dependent release of gliotransmitters. Furthermore, astrocytes constitute the main uptake system for glutamate, contribute to potassium spatial buffering, as well as to GABA clearance. These cells therefore constantly monitor synaptic activity, and are thereby sensitive indicators for alterations in synaptically-released glutamate, GABA and extracellular potassium levels. Additionally, alterations in astroglial uptake activity or buffering capacity can have severe effects on neuronal functions, and might be overlooked when characterizing physiopathological situations or knockout mice. Dual recording of neuronal and astroglial activities is therefore an important method to study alterations in synaptic strength associated to concomitant changes in astroglial uptake and buffering capacities. Here we describe how to prepare hippocampal slices, how to identify stratum radiatum astrocytes, and how to record simultaneously neuronal and astroglial electrophysiological responses. Furthermore, we describe how to isolate pharmacologically the synaptically-evoked astroglial currents.

The preparation of acute brain slices from isolated hippocampi, as well as the simultaneous electrophysiological recordings of astrocytes and neurons in stratum radiatum during stimulation of schaffer collaterals is described. The pharmacological isolation of astroglial potassium and glutamate transporter currents is demonstrated.

双突触诱发的星形胶质细胞和神经元的反应，在急性海马脑片电生理记录

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs ...

A Computer-assisted Multi-electrode Patch-clamp System

The patch-clamp technique is today the most well-established method for recording electrical activity from individual neurons or their subcellular compartments. Nevertheless, achieving stable recordings, even from individual cells, remains a time-consuming procedure of considerable complexity. Automation of many steps in conjunction with efficient information display can greatly assist experimentalists in performing a larger number of recordings with greater reliability and in less time. In order to achieve large-scale recordings we concluded the most efficient approach is not to fully automatize the process but to simplify the experimental steps and reduce the chances of human error while efficiently incorporating the experimenter's experience and visual feedback. With these goals in mind we developed a computer-assisted system which centralizes all the controls necessary for a multi-electrode patch-clamp experiment in a single interface, a commercially available wireless gamepad, while displaying experiment related information and guidance cues on the computer screen. Here we describe the different components of the system which allowed us to reduce the time required for achieving the recording configuration and substantially increase the chances of successfully recording large numbers of neurons simultaneously.

Multi-electrode patch-clamp recordings constitute a complex task. Here we show how, by automating of many of the experimental steps, it is possible to accelerate the process leading to qualitative improvement in performance and number of recordings.

计算机辅助多电极膜片钳系统

The patch-clamp technique is today the most  well-established method for recording electrical activity from individual  neurons or their subcellular ...

Direct-current Stimulation and Multi-electrode Array Recording of Seizure-like Activity in Mice Brain Slice Preparation

Cathodal transcranial direct-current stimulation (tDCS) induces suppressive effects on drug-resistant seizures. To perform effective actions, the stimulation parameters (e.g., orientation, field strength, and stimulation duration) need to be examined in mice brain slice preparations. Testing and arranging the orientation of the electrode relative to the position of the mice brain slice are feasible. The present method preserves the thalamocingulate pathway to evaluate the effect of DCS on anterior cingulate cortex seizure-like activities. The results of the multichannel array recordings indicated that cathodal DCS significantly decreased the amplitude of the stimulation-evoked responses and duration of 4-aminopyridine and bicuculline-induced seizure-like activity. This study also found that cathodal DCS applications at 15 min caused long-term depression in the thalamocingulate pathway. The present study investigates the effects of DCS on thalamocingulate synaptic plasticity and acute seizure-like activities. The current procedure can test the optimal stimulation parameters including orientation, field strength, and stimulation duration in an in vitro mouse model. Also, the method can evaluate the effects of DCS on cortical seizure-like activities at both the cellular and network levels.

Studies have shown that cathodal transcranial direct-current stimulation can produce suppressive effects on drug-resistant seizures. In this study, an in vitro experimental setup was devised in which the direct-current stimulation and multielectrode array recording of seizure-like activity were evaluated in mice brain slice preparation. The direct-current stimulation parameters were evaluated.

小鼠脑切片准备扣押样活性的直接电流刺激和多电极阵列记录

Cathodal transcranial direct-current stimulation (tDCS) induces suppressive effects on drug-resistant seizures. To perform effective actions, the ...

Spiral Ganglion Neuron Explant Culture and Electrophysiology on Multi Electrode Arrays

Spiral ganglion neurons (SGNs) participate in the physiological process of hearing by relaying signals from sensory hair cells to the cochlear nucleus in the brain stem. Loss of hair cells is a major cause of sensory hearing loss. Prosthetic devices such as cochlear implants function by bypassing lost hair cells and directly stimulating SGNs electrically, allowing for restoration of hearing in deaf patients. The performance of these devices depends on the functionality of SGNs, the implantation procedure and on the distance between the electrodes and the auditory neurons. 
We hypothesized, that reducing the distance between the SGNs and the electrode array of the implant would allow for improved stimulation and frequency resolution, with the best results in a gapless position. Currently we lack in vitro culture systems to study, modify and optimize the interaction between auditory neurons and electrode arrays and characterize their electrophysiological response. To address these issues, we developed an in vitro bioassay using SGN cultures on a planar multi electrode array (MEA). With this method we were able to perform extracellular recording of the basal and electrically induced activity of a population of spiral ganglion neurons. We were also able to optimize stimulation protocols and analyze the response to electrical stimuli as a function of the electrode distance. This platform could also be used to optimize electrode features such as surface coatings.

We present a protocol to culture primary murine spiral ganglion neuron explants on multi electrode arrays to study neuronal response profiles and optimize stimulation parameters. Such studies aim to improve the neuron-electrode interface of cochlear implants to benefit hearing in patients as well as the energy consumption of the device.

螺旋神经节神经元外植体培养及电多电极阵列

Spiral ganglion neurons (SGNs) participate in the physiological process of hearing by relaying signals from sensory hair cells to the cochlear nucleus in ...

A Visual Guide to Sorting Electrophysiological Recordings Using 'SpikeSorter'

Few stand-alone software applications are available for sorting spikes from recordings made with multi-electrode arrays. Ideally, an application should be user friendly with a graphical user interface, able to read data files in a variety of formats, and provide users with a flexible set of tools giving them the ability to detect and sort extracellular voltage waveforms from different units with some degree of reliability. Previously published spike sorting methods are now available in a software program, SpikeSorter, intended to provide electrophysiologists with a complete set of tools for sorting, starting from raw recorded data file and ending with the export of sorted spikes times. Procedures are automated to the extent this is currently possible. The article explains and illustrates the use of the program. A representative data file is opened, extracellular traces are filtered, events are detected and then clustered. A number of problems that commonly occur during sorting are illustrated, including the artefactual over-splitting of units due to the tendency of some units to fire spikes in pairs where the second spike is significantly smaller than the first, and over-splitting caused by slow variation in spike height over time encountered in some units. The accuracy of SpikeSorter's performance has been tested with surrogate ground truth data and found to be comparable to that of other algorithms in current development.

The article shows how to use the program SpikeSorter to detect and sort spikes in extracellular recordings made with multi-electrode arrays.

可视化指南排序的电生理记录使用"SpikeSorter"

Few stand-alone software applications are available for sorting spikes from recordings made with multi-electrode arrays. Ideally, an application should be ...

Whole-cell Patch-clamp Recordings for Electrophysiological Determination of Ion Selectivity in Channelrhodopsins

Over the past decade, channelrhodopsins became indispensable in neuroscientific research where they are used as tools to non-invasively manipulate electrical processes in target cells. In this context, ion selectivity of a channelrhodopsin is of particular importance. This article describes the investigation of chloride selectivity for a recently identified anion-conducting channelrhodopsin of Proteomonas sulcata via electrophysiological patch-clamp recordings on HEK293 cells. The experimental procedure for measuring light-gated photocurrents demands a fast switchable &#8211; ideally monochromatic &#8211; light source coupled into the microscope of an otherwise conventional patch-clamp setup. Preparative procedures prior to the experiment are outlined involving preparation of buffered solutions, considerations on liquid junction potentials, seeding and transfection of cells, and pulling of patch pipettes. The actual recording of current-voltage relations to determine the reversal potentials for different chloride concentrations takes place 24 h to 48 h after transfection. Finally, electrophysiological data are analyzed with respect to theoretical considerations of chloride conduction.

This article describes how the ion selectivity of channelrhodopsin is determined with electrophysiological whole-cell patch-clamp recordings using HEK293 cells. Here, the experimental procedure for investigating chloride selectivity of an anion-selective channelrhodopsin is demonstrated. However, the procedure is transferable to other channelrhodopsins of distinct selectivity.

全细胞贴片记录用于电泳生理测定通道视紫质中的离子选择性

Over the past decade, channelrhodopsins became indispensable in neuroscientific research where they are used as tools to non-invasively manipulate ...

Zebrafish In Situ Spinal Cord Preparation for Electrophysiological Recordings from Spinal Sensory and Motor Neurons

Zebrafish, first introduced as a developmental model, have gained popularity in many other fields. The ease of rearing large numbers of rapidly developing organisms, combined with the embryonic optical clarity, served as initial compelling attributes of this model. Over the past two decades, the success of this model has been further propelled by its amenability to large-scale mutagenesis screens and by the ease of transgenesis. More recently, gene-editing approaches have extended the power of the model.
For neurodevelopmental studies, the zebrafish embryo and larva provide a model to which multiple methods can be applied. Here, we focus on methods that allow the study of an essential property of neurons, electrical excitability. Our preparation for the electrophysiological study of zebrafish spinal neurons involves the use of veterinarian suture glue to secure the preparation to a recording chamber. Alternative methods for recording from zebrafish embryos and larvae involve the attachment of the preparation to the chamber using a fine tungsten pin1,2,3,4,5. A tungsten pin is most often used to mount the preparation in a lateral orientation, although it has been used to mount larvae dorsal-side up4. The suture glue has been used to mount embryos and larvae in both orientations. Using the glue, a minimal dissection can be performed, allowing access to spinal neurons without the use of an enzymatic treatment, thereby avoiding any resultant damage. However, for larvae, it is necessary to apply a brief enzyme treatment to remove the muscle tissue surrounding the spinal cord. The methods described here have been used to study the intrinsic electrical properties of motor neurons, interneurons, and sensory neurons at several developmental stages6,7,8,9.

Zebrafish In Situ Spinal Cord Preparation for Electrophysiological Recordings from Spinal Sensory and Motor Neurons

This manuscript describes methods for electrophysiological recordings from spinal neurons of zebrafish embryos and larvae. The preparation maintains neurons in situ and often involves minimum dissection. These methods allow for the electrophysiological study of a variety of spinal neurons, from the initial electrical excitability acquisition through the early larval stages.

斑马鱼<em&gt;原位</em&gt;脊髓制备用于脊髓感觉和运动神经元的电生理记录

Zebrafish, first introduced as a developmental model, have gained popularity in many other fields. The ease of rearing large numbers of rapidly developing ...