这项工作是由酯和约瑟夫·麦克奈特Klingenstein基金，捐赠基金，NINDS 1R01NS066408-01A1。

1。组件概述<ol><li>一个完整的微硬盘组成的几个主要部件（ 图1）:作为上层建筑具有精细螺纹输出轴的驱动器，电动机，带有电极和提供了一个电连接的点的螺纹班车的机箱，以及一个Omnetics（或同等学历）连接器。 </li><li>底盘，电极班车，电极穿梭管的自定义组件的设计（SolidWorks的三维CAD软件）和加工的精密加工。其它所有组件都是市售的。 </li><li>底盘和穿梭从轻量级的聚醚酰亚胺（PEI），而电极班车管切割成一定的尺寸从不锈钢皮下注射管（0.0293"外径×0.00975"ID）用车床加工。虽然这些组分可以由其它材料切断，我们已经发现，PEI和不锈钢具有适当的平衡的可加工性，强度，和w8我们的应用程序。 </li><li>为了组装的驱动器，马达控制器，能够纺纱的低直流电压的步进电机的，是必需的。我们采用基于三相电机驱动器IC（冯哈伯BLD05002可以从制造商）结合的时间步长输入的IC定时器，提供定制的解决方案。多种的，现成的解决方案，也可用（ 例如，冯哈伯MCBL05002或萨特仪器MP-285）。 </li></ol> 2。驱动器机箱的制备和组装<ol><li>以优良的对角线切割机，修剪底部的Omnetics（或同等学历）连接器的插针- 1.5毫米，1毫米的后排前排（ 图2A）。为了提高驱动机架和连接器之间的环氧键的强度，用手术刀以粗糙化后表面的两个组成部分。 </li><li>使用手术刀，切四聚酰亚胺管（0.0113"（内径）×0.0133 OD）15毫米长的部分用于线区集成开发环境。小心地固定在机箱中的一小副的开放腔的下降（ 图2B）。使用氰基丙烯酸酯胶，聚酰亚胺管连接到机箱。他们应该奠定与机箱后部的表面齐平，并面向彼此平行地，如在图2B中所示。 </li><li>一旦干燥，修剪的聚酰亚胺管，以约6毫米的长度和删除的废物部。其余的管应该是从机箱的底部边缘约3毫米。 </li><li>与探头或电缆的配合，以确保适当的间隙，连接器应安装有几毫米以上的聚酰亚胺管和平行与机箱表面（ 图2B）。使用环氧树脂（托盖章）建立一个小底座，连接器，将其放在最上面的针脚排与短，周边连接器添加更多的环氧树脂。不要继续进行，直至环氧树脂完全固化。 </li><li>卸下机箱从副，转移到连接到电机控制器（ 即 ，在我们的例子中，允许的夹具，该夹具是一个简单的刚性杆与的的交配Omnetics接头固定到驱动器的前表面朝上的一端）和定向。引导电机线孔与机箱和滑动电机在底座上的凹槽。电机和驱动轴应该是平行的机箱，和适合紧贴地对底部和侧面。 </li><li>环氧树脂的电动机机箱，照顾，不阻碍驱动轴或（见乇密封放置在图4中）的空间，将被安装在电极。允许环氧树脂，固化。 </li><li>旋转机箱上，使连接器是可访问的。为了支持的电机电线和防止意外破损，加一小滴的快速设定环氧树脂，他们从机箱中出现的电线。修剪，带材和焊接导线连接器上的相应的触点（ 图2A）。测试电机确保在两个方向上的轴旋转平稳。 </li></ol> 3。电极班车大会<ol><li>为了尽量减少运动引起的工件在录音的潜力，它是必不可少的电极班车适合紧贴在机箱中。测试适合的穿梭通过螺纹上的电机轴和运行它向上和向下的整个长度的轴在低速。要小心，不要楔入穿梭在驱动器中，因而在扭矩马达，这是很容易在这个阶段，永久损坏塑料减速机，。如果先前旋转的电机轴停止转动，减速机可能被损坏。 </li><li>如果航天飞机不能平稳地运行下来的轴，检查底盘表面和驱动螺杆碎片。用细镊子或压缩空气，外套的线程在一些轻质矿物油和复检的穿梭配合中删除。 </li><li>如果航天飞机是太紧张了，用钳子和细晶粒磨刀石，以消除阻碍材料来回在侧的班车。小心均匀地去除材料。 </li><li>如果航天飞机是过于宽松，使得它的倾斜和聊友在旅行期间，胶水的小条子的班车，填补了国内空白，它与机箱表面的透明薄膜。 </li><li>一旦航天飞机已经好了，取出梭轴，然后按一个的不锈钢班车管到每四个大孔中穿梭。每个应居中航天飞机上的与所有四个管冲洗（ 图3）的端部的。确保他们与氰基丙烯酸酯胶施加到该管的基极一小滴。要非常小心，不要让胶水的边缘，穿梭或进入管。 </li><li>的主题完成电极班车驱动轴，并把它一路下跌到基本的线程。 </li></ol> 4。导管和电极安装<ol><li>切4个25毫米长的段的聚酰亚胺tubin的克（0.0045"（内径）×0.006"OD）使用作为电极导向件。此外，切四电极至40毫米作为虚拟电极将保持导管的位置在装配过程中使用的部分。 （请注意，这些虚设电极仅供组件和用于记录将不会被使用，所以为每个组件，则可以重复使用相同的。）上滑动管的每个部分上的虚设电极，使得电极延伸〜10毫米超出在管的端部。 </li><li>安排每个的聚酰亚胺管在机箱中，如在图4A中示出。这将导致在所述电极导板的端部，彼此齐平，并对齐，在电动机轴的基础上，朝向电极穿梭张开的相对端部，在该驱动器的底部会聚。佳铸铁中混入少量和应用的电极导板的上端约2毫米以下。允许干燥。 </li><li>移除虚设电极和重新定位的自由端部的电极引导等它们形成一个捆紧在驱动器的底部。使用细金属丝，以保持他们在地方暂时（ 图4B），它可能会有所帮助。修正KWIK-主演的新职位。允许干燥。 </li><li>用一把锋利的手术刀，修剪它们超出微硬盘的底部电极导管。在植入时，这些管将引导的驱动器的底部，（搁在颅骨）从电极表面的大脑。因此，在该管应修剪的长度取决于的解剖结构上的植入位点（ 图5）。燕雀，这是约1.5毫米。 </li><li>要连接的电极的连接器，切断一个30毫米长的边的绝缘铂丝（0.003"直径）和钢带1 mm的绝缘层，从一端焊接到连接器上的信号引脚之一。弯曲的线下的连接器和将其推过的金属丝导管之一。拉出丝嘲讽和环绕在t的导槽他在机箱顶部。 </li><li>用剪刀或细金属丝的切割机，切割到〜25毫米长的电极。照顾做到这一点，而不会弯曲的电极或损坏的前端;如果任一发生，电极不能使用。随着现在电极班车，在所述轴的顶部上，插入到聚酰亚胺管的电极，并把它通过不锈钢管中穿梭。将它放置，使得在电极的前端是平齐的聚酰亚胺管的底端。修剪穿梭管的电极1毫米以上。 </li><li>用细集钳子，剥去绝缘层的最后2毫米的电极和最后2毫米的铂丝。重新放置电极在航天飞机管，插入铂丝管，两针一起滑短1毫米长的钨丝（0.008"直径）的管，这应该是一个紧密配合，将同时提供一个机械的和之间的电连接的电极和所述导线（ 图3）。 </li><li>运行穿梭向上和向下的长度，以确保续运动的驱动轴。 </li><li>重复4.5-4.8，直到所有已安装电极。 </li><li>切割一个30毫米长的边的银线（0.005"直径）使用作为地线。剥去1 mm的绝缘层的接地接脚的一端和焊料。 </li></ol> 5。最后的组装和胚胎植入前的准备<ol><li>为了减少对导向管的电极的摩擦，填充导向管与轻质矿物油。毛细管作用是足以填满管放置在一端从一滴油。 </li><li>要建立一个保护盖用于电机和电极，切成12毫米×25毫米的矩形的透明度，并把它折叠，用于广泛的马蹄形。使用腈基丙烯酸酯胶盖机箱的外表面（ 图6）。切割15毫米x 6毫米矩形的透明度和使用氰基丙烯酸酯胶周边连接器。为了防止干扰ENCE透明度的长边与连接器，应该是与连接器的顶部边缘平齐。 </li><li>在植入前，消毒的电极和导向管通过降低扩展到10:1的去离子水和10分钟的漂白剂溶液的电极。然后，用无菌水彻底冲洗。 </li><li>步骤2.5中所使用的类似的夹具可用于保存在立体机械手在植入过程中的微硬盘。 </li></ol> 6。代表性的成果此协议需要大约5个小时的手，用环氧树脂和胶水干6-8小时穿插的装配时间。然而，一旦微硬盘已组装的第一时间，它可以制备以供重复使用（ 即 ，电极，电极导管，并可以更换电极布线）小于2小时。遵循这个协议的录音品质和性格，当然，是部分依赖于记录目标，在选择的电极，并在探头或进一步上游的微硬盘进行任何处理。这一边，从行为动物超过超过十个星期的时间跨度，因此能够获得稳定的录音。古纹状体栎（RA）中的行为鸣禽鲁棒核的单个单元录音从图7A中所示的实施例， 图7B中所示的多单元来自同一区域的活动。 <img alt="图1" src="/files/ftp_upload/4314/4314fig1.jpg" /> 图1的组装的机动微硬盘的3D模型。微硬盘由几个主要部分组成:（一）机箱，（二）螺纹输出轴电机，（三）电极穿梭，（四）穿梭管，（V）的电极，电极导管（六），（七）连接器。 <img alt="图2" src="/files/ftp_upload/4314/4314fig2.jpg" /> 图2 A）。型号和连接器的连接图。马达线:B，I，L。电极导线:C，D，E，F。接地线:K。所有其它引脚可用于辅助仪器。 B）照片的连接器连接到机箱上托密封环氧树脂。注意连接器和底层的金属丝导管的位置。清晰和，因此，在位置干燥的腈基丙烯酸酯胶保持在导向管是在图像中不可见的。 <img alt="图3" src="/files/ftp_upload/4314/4314fig3.jpg" /> 图3。装配电极穿梭。 <img alt="图4" src="/files/ftp_upload/4314/4314fig4.jpg" /> 图4 A）的电极导管定位在机箱中之前施加佳-施法。 B）为了确保电极退出中的驱动器底部平行，带导向管的底端，成捆紧，然后再加入KWIK-主演的最后一滴。 图5，电极引导管的量超出该驱动器的底部延伸指定的植入部位的解剖结构。该图说明了用于确定导管的长度相关的尺寸。用于植入在鸣禽，1.5毫米就足够了。 <img alt="图6" src="/files/ftp_upload/4314/4314fig6.jpg" /> 图6。完成的电动微硬盘防护罩连接。 <img alt="图7" src="/files/ftp_upload/4314/4314fig7.jpg" /> 图7。强大的古纹状体栎核（RA）在斑胸草雀的行为代表的录音。 A）10MΩ铂铱电极记录单单位活动。上图:例如记录了一个星期后植入。例如下:从相同的电极记录如上述，购买9周。 B）多单元的活动记录与1MΩ铂电极。 <a href="http://www.jove.com/files/ftp_upload/4314/4314fig7large.jpg" target="_blank">点击此处查看大图</a> 。 其他文件:2010年与SolidWorks CAD软件设计的底盘，班车服务和电极管。这部分文件提供了专有的（*。SLDPRT）和供应商中立（IGES）的格式和尺寸的图纸生产以PDF格式提供。

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没有利益冲突的声明。

这里介绍的协议将导致一个能够以最小的运动伪影的高品质的录音设备，只有采取适当的照顾与建设。适合穿梭在机箱的如果至关重要太紧张，电机过载的风险是高过于宽松，显着的运动伪影的风险是高。一个理想的选择，可以让穿梭旅行的整个长度的螺纹轴的位置或抖动没有倾斜了。 记录电极的选择同样是重要的;材料，阻抗，绝缘，和小费档案中的选择可能会影响组织的?...

<table border="1"&gt;<tbody&gt;<tr&gt;<td<strong&gt;项目名称</strong</td&gt;<td<strong&gt;公司</strong</td&gt;<td<strong&gt;产品目录号</strong</td&gt;<td<strong&gt;评论</strong</td&gt;</tr&gt;<tr&gt;<td&gt;机壳</td&gt;<td&gt;自定义</td&gt;<td</td&gt;<td从PEI&gt;"剪切"</td&gt;</tr&gt;<tr&gt;<td&gt;电极班车</td&gt;<td&gt;自定义</td&gt;<td</td&gt;<td从PEI&gt;"剪切"</td&gt;</tr&gt;<tr&gt;<td&gt;接送管</td&gt;<td&gt;自定义</td&gt;<td</td&gt;<td剪切不锈钢</td&gt;</tr&gt;<tr&gt;<td&gt;连接​​器</td&gt;<td&gt; Omnetics</td&gt;<td&gt; A7886-001</td&gt;<td配合A7877-001</td&gt;</tr&gt;<tr&gt;<td&gt;电机/减速机</td&gt;<td&gt;冯哈伯</td&gt;<td&gt; 0206-A-001-B-021-47:1</td&gt;<td</td&gt;</tr&gt;<tr&gt;<td&gt;电线指南</td&gt;<td&gt;小零件公司</td&gt;<td&gt; SWPT-0113-12</td&gt;<td</td&gt;</tr&gt;<tr&gt;<td&gt;电极指南</td&gt;<td&gt;小零件公司</td&gt;<td&gt; SWPT-0045-12</td&gt;<td</td&gt;</tr&gt;<tr&gt;<td&gt;10MΩ铂铱电极</td&gt;<td&gt;柱石，公司</td&gt;<td&gt; PI2PT310.0H3</td&gt;<td</td&gt;</tr&gt;<tr&gt;<td&gt;铂金导丝</td&gt;<td&gt; AM系统</td&gt;<td&gt; 772000</td&gt;<td电极线</td&gt;</tr&gt;<tr&gt;<td&gt;银线</td&gt;<td&gt; AM系统</td&gt;<td&gt; 786000</td&gt;<td对于地线</td&gt;</tr&gt;<tr&gt;<td&gt;钨丝</td&gt;<td&gt; AM系统</td&gt;<td&gt; 797000</td&gt;<td&gt;电极针</td&gt;</tr&gt;<tr&gt;<td&gt;透明度</td&gt;<td&gt; 3M</td&gt;<td&gt; AF4300</td&gt;<td</td&gt;</tr&gt;<tr&gt;<td乇封</td&gt;<td&gt;瓦里安</td&gt;<td&gt; 9530001</td&gt;<td</td&gt;</tr&gt;<tr&gt;<tdKWIK-主演</td&gt;<td&gt; WPI</td&gt;<tdKWIK-CAST</td&gt;<td</td&gt;</tr&gt;<tr&gt;<td&gt;氰基丙烯酸酯</td&gt;<td&gt; KRAZY胶</td&gt;<td&gt; KG517</td&gt;<td</td&gt;</tr&gt;<tr&gt;<td&gt;快速设置环氧</td&gt;<td&gt;哈德曼</td&gt;<td&gt; 04001</td&gt;<td</td&gt;</tr&gt;<tr&gt;<td&gt;轻质矿物油</td&gt;<td&gt; SIGMA</td&gt;<td&gt; M5310</td&gt;<td</td&gt;</tr&gt;<tr&gt;<td&gt;氯漂白剂</td&gt;<td</td&gt;<td</td&gt;<td</td&gt;</tr&gt;<tr&gt;<td&gt;对角线切割机</td&gt;<td</td&gt;<td</td&gt;<td</td&gt;</tr&gt;<tr&gt;<td&gt;手术刀</td&gt;<td</td&gt;<td</td&gt;<td</td&gt;</tr&gt;<tr&gt;<td&gt;钳</td&gt;<td</td&gt;<td</td&gt;<td</td&gt;</tr&gt;<tr&gt;<td&gt;驱动器夹具</td&gt;<td&gt;自定义</td&gt;<td</td&gt;<td&gt;环氧交配连接器，注射器或棒</td&gt;</tr&gt;<tr&gt;<td&gt;小副</td&gt;<td</td&gt;<td</td&gt;<td</td&gt;</tr&gt;</tbody&gt;</table

design and assembly of an ultra-light motorized microdrive for chronic neural recordings in small animals

的能力，长期记录神经元在自由活动的动物的种群已被证明是非常宝贵的工具，用于解剖的神经回路的功能相关的各种自然行为，包括导航1，决策2,3，和上一代的复杂的运动序列4 ，5,6。在精密加工方面的进展已可用于制造重量轻的设备，适合慢性录音的小动物，如小鼠和鸣禽。调整电极的位置与远程控制电机的能力进一步提高各种行为背景的记录产量的减少动物的处理6,7。 在这里，我们描述了一个协议，以建立一个超轻型电动微硬盘为长期的慢性记录的小动物。我们的设计是从早前公布的第7版，已被改编为易用性和成本有效的veness更加实用和广泛的研究人员访问。这种成熟的设计8,9,10,11允许的精细电极，远程定位〜5毫米的范围内，重量不超过750毫克时完全组装。我们提出如何构建和组装这些驱动器，包括三维CAD图纸的所有自定义的微硬盘组件的完整的协议。

Watch this Scientific Journal Video about Design and Assembly of an Ultra-light Motorized Microdrive for Chronic Neural Recordings in Small Animals at JoVE.com

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

一个超轻型电动微硬盘的设计，制造和装配。该设备提供了成本效益和易于使用的解决方案，单台小行为的动物慢性录音。

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 ...

design-assembly-an-ultra-light-motorized-microdrive-for-chronic

Research

JoVE Journal

Neuroscience

12.0K 次观看.  Harvard University. 一个超轻型电动微硬盘的设计，制造和装配。该设备提供了成本效益和易于使用的解决方案，单台小行为的动物慢性录音。

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

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

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

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

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

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

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

科研

神经科学

A Thin-skull Window Technique for Chronic Two-photon In vivo Imaging of Murine Microglia in Models of Neuroinflammation

Traditionally in neuroscience, in vivo two photon imaging of the murine central nervous system has either involved the use of open-skull1,2 or thinned-skull 3 preparations. While the open-skull technique is very versatile, it is not optimal for studying microglia because it is invasive and can cause microglial activation. Even though the thinned-skull approach is minimally invasive, the repeated re-thinning of skull required for chronic imaging increases the risks of tissue injury and microglial activation and allows for a limited number of imaging sessions. Here we present a chronic thin-skull window method for monitoring murine microglia in vivo over an extended period of time using two-photon microscopy. We demonstrate how to prepare a stable, accessible, thinned-skull cortical window (TSCW) with an apposed glass coverslip that remains translucent over the course of three weeks of intermittent observation. This TSCW preparation is far more immunologically inert with respect to microglial activation than open craniotomy or repeated skull thinning and allows an arbitrary number of imaging sessions during a time period of weeks. We prepare TSCW in CX3CR1 GFP/+ mice 4 to visualize microglia with enhanced green fluorescent protein to &le;150 &mu;m beneath the pial surface. We also show that this preparation can be used in conjunction with stereotactic brain injections of the HIV-1 neurotoxic protein Tat, adjacent to the TSCW, which is capable of inducing durable microgliosis. Therefore, this method is extremely useful for examining changes in microglial morphology and motility over time in the living brain in models of HIV Associated Neurocognitive Disorder (HAND) and other neurodegenerative diseases with a neuroinflammatory component.

A Thin-skull Window Technique for Chronic Two-photon In vivo Imaging of Murine Microglia in Models of Neuroinflammation

We describe a method for repeatedly visualizing murine microglia and circulating monocytes in vivo over hours, days or weeks using transcranial two-photon microscopy. We demonstrate how to prepare a thinned-skull window that allows intermittent observation of quiescent microglia that can be activated by adjacent stereotactic injection of the HIV-1 regulatory protein Tat.

一个慢性双光子薄颅骨窗口技术在体内 Neuroinflammation模型小鼠小胶质细胞成像

Traditionally in neuroscience, in vivo two photon imaging of the murine central nervous system has either involved the use of open-skull1,2 or ...

Combining Behavioral Endocrinology and Experimental Economics: Testosterone and Social Decision Making

Behavioral endocrinological research in humans as well as in animals suggests that testosterone plays a key role in social interactions. Studies in rodents have shown a direct link between testosterone and aggressive behavior1 and folk wisdom adapts these findings to humans, suggesting that testosterone induces antisocial, egoistic or even aggressive behavior2. However, many researchers doubt a direct testosterone-aggression link in humans, arguing instead that testosterone is primarily involved in status-related behavior3,4. As a high status can also be achieved by aggressive and antisocial means it can be difficult to distinguish between anti-social and status seeking behavior.
We therefore set up an experimental environment, in which status can only be achieved by prosocial means. In a double-blind and placebo-controlled experiment, we administered a single sublingual dose of 0.5 mg of testosterone (with a hydroxypropyl-&beta;-cyclodextrin carrier) to 121 women and investigated their social interaction behavior in an economic bargaining paradigm. Real monetary incentives are at stake in this paradigm; every player A receives a certain amount of money and has to make an offer to another player B on how to share the money. If B accepts, she gets what was offered and player A keeps the rest. If B refuses the offer, nobody gets anything. A status seeking player A is expected to avoid being rejected by behaving in a prosocial way, i.e. by making higher offers.
The results show that if expectations about the hormone are controlled for, testosterone administration leads to a significant increase in fair bargaining offers compared to placebo. The role of expectations is reflected in the fact that subjects who report that they believe to have received testosterone make lower offers than those who say they believe that they were treated with a placebo. These findings suggest that the experimental economics approach is sensitive for detecting neurobiological effects as subtle as those achieved by administration of hormones. Moreover, the findings point towards the importance of both psychosocial as well as neuroendocrine factors in determining the influence of testosterone on human social behavior.

The procedure described in this protocol shows that testosterone administration and folk beliefs about testosterone may be associated with directly opposed social behaviors.

睾酮和社会决策，结合行为内分泌和实验经济学:

Behavioral endocrinological research in humans as well as in animals suggests that testosterone plays a key role in social interactions. Studies in ...

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

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

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

高密度的自由移动，使用基于聚酰亚胺的微电极的小鼠的脑电图记录

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

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 Procedure for Implanting Organized Arrays of Microwires for Single-unit Recordings in Awake, Behaving Animals

In vivo electrophysiological recordings in the awake, behaving animal provide a powerful method for understanding neural signaling at the single-cell level. The technique allows experimenters to examine temporally and regionally specific firing patterns in order to correlate recorded action potentials with ongoing behavior. Moreover, single-unit recordings can be combined with a plethora of other techniques in order to produce comprehensive explanations of neural function. In this article, we describe the anesthesia and preparation for microwire implantation. Subsequently, we enumerate the necessary equipment and surgical steps to accurately insert a microwire array into a target structure. Lastly, we briefly describe the equipment used to record from each individual electrode in the array. The fixed microwire arrays described are well-suited for chronic implantation and allow for longitudinal recordings of neural data in almost any behavioral preparation. We discuss tracing electrode tracks to triangulate microwire positions as well as ways to combine microwire implantation with immunohistochemical techniques in order to increase the anatomical specificity of recorded results.

Implanting organized arrays of microwires for use in single-unit electrophysiological recordings presents a number of technical challenges.&#160;Methods for performing this technique and the equipment necessary are described.&#160;Also, the beneficial use of organized microwire arrays to record from distinct neural subregions with high spatial selectivity is discussed.

一个程序植入微丝的有组织阵列的单机录音清醒，举止动物

In vivo electrophysiological recordings in the awake, behaving animal provide a powerful method for understanding neural signaling at the single-cell ...

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

The number of physiological investigations in the mouse, mus musculus, has experienced a recent surge, paralleling the growth in methods of genetic targeting for microcircuit dissection and disease modeling. The introduction of optogenetics, for example, has allowed for bidirectional manipulation of genetically-identified neurons, at an unprecedented temporal resolution. To capitalize on these tools and gain insight into dynamic interactions among brain microcircuits, it is essential that one has the ability to record from ensembles of neurons deep within the brain of this small rodent, in both head-fixed and freely behaving preparations. To record from deep structures and distinct cell layers requires a preparation that allows precise advancement of electrodes towards desired brain regions. To record neural ensembles, it is necessary that each electrode be independently movable, allowing the experimenter to resolve individual cells while leaving neighboring electrodes undisturbed. To do both in a freely behaving mouse requires an electrode drive that is lightweight, resilient, and highly customizable for targeting specific brain structures.
A technique for designing and fabricating miniature, ultralight weight, microdrive electrode arrays that are individually customizable and easily assembled from commercially available parts is presented. These devices are easily scalable and can be customized to the structure being targeted; it has been used successfully to record from thalamic and cortical regions in a freely behaving animal during natural behavior.

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 ...

Simultaneous Recordings of Cortical Local Field Potentials, Electrocardiogram, Electromyogram, and Breathing Rhythm from a Freely Moving Rat

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls body functions and internal organ rhythms when animals are exposed to emotional challenges and changes in their living environments. In general experiments, signals from different organs, such as the brain and the heart, are recorded by independent recording systems that require multiple recording devices and different procedures for processing the data files. This study describes a new method that can simultaneously monitor electrical biosignals, including tens of local field potentials in multiple brain regions, electrocardiograms that represent the cardiac rhythm, electromyograms that represent awake/sleep-related muscle contraction, and breathing signals, in a freely moving rat. The recording configuration of this method is based on a conventional micro-drive array for cortical local field potential recordings in which tens of electrodes are accommodated, and the signals obtained from these electrodes are integrated into a single electrical board mounted on the animal's head. Here, this recording system was improved so that signals from the peripheral organs are also transferred to an electrical interface board. In a single surgery, electrodes are first separately implanted into the appropriate body parts and the target brain areas. The open ends of all of these electrodes are then soldered to individual channels of the electrical board above the animal's head so that all of the signals can be integrated into the single electrical board. Connecting this board to a recording device allows for the collection of all of the signals into a single device, which reduces experimental costs and simplifies data processing, because all data can be handled in the same data file. This technique will aid the understanding of the neurophysiological correlates of the associations between central and peripheral organs.

This study introduces a method for the simultaneous recording of local field potentials in the brain, electrocardiograms, electromyograms, and breathing signals of a freely moving rat. This technique, which reduces experimental costs and simplifies data analysis, will contribute to the understanding of the interactions between the brain and peripheral organs.

自由运动大鼠皮质局部电位、心电图、肌电图和呼吸节律的同步记录

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls ...

Chronic Implantation of Whole-cortical Electrocorticographic Array in the Common Marmoset

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent development of thin, flexible ECoG electrodes has enabled conduction of stable recordings of large-scale cortical activity. We have developed a whole-cortical ECoG array for the common marmoset. The array continuously covers almost the entire lateral surface of cortical hemisphere, from the occipital pole to the temporal and frontal poles, and it captures whole-cortical neural activity in one shot. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain. Marmosets have two advantages regarding ECoG recordings, one being the homologous organization of anatomical structures in humans and macaques, including frontal, parietal, and temporal complexes. The other advantage is that the marmoset brain is lissencephalic and contains a large number of complexes, which are more difficult to access in macaques with ECoG, that are exposed to the brain surface.These features allow direct access to most cortical areas beneath the surface of the brain. This system provides an opportunity to investigate global cortical information processing with high resolutions at a sub-millisecond order in time and millimeter order in space.

We have developed a whole-cortical electrocorticographic array for the common marmoset that continuously covers almost the entire lateral surface of cortex, from the occipital pole to the temporal and frontal poles. This protocol describes a chronic implantation procedure of the array in the epidural space of the marmoset brain.

常见马莫塞的全皮质电皮质阵列的慢性植入

Electrocorticography (ECoG) allows the monitoring of electrical field potentials from the cerebral cortex with high spatiotemporal resolution. Recent ...

Rabies Necropsy Techniques in Large and Small Animals

The New York State Department of Health (NYSDOH) Rabies Laboratory receives between 6,000 to 9,000 specimens annually and performs rabies testing for the entire state, with the exception of New York City. The Rabies laboratory necropsies a variety of animals ranging in size from bats to bovids. Most of these specimens are animals exhibiting neurological signs, however, less than 10% actually test positive for rabies; implying trauma, lesions or other infectious agents as the cause of these symptoms. Due to the risk of aerosolizing undiagnosed infectious agents, the Rabies Laboratory does not use power tools or saws. Three necropsy techniques will be presented for animals whose skulls are impenetrable with scissors. The laboratory has implemented these techniques to decrease potential exposure to infectious agents, eliminate unnecessary manipulation of the specimen and reduce processing time. The advantages of a preferred technique opposed to another is subject to the trained individual processing the specimen.

The goal of this protocol is to demonstrate safe necropsy techniques in small and large animals to obtain satisfactory tissue samples for rabies testing.

大、小动物狂犬病的解剖技术

The New York State Department of Health (NYSDOH) Rabies Laboratory receives between 6,000 to 9,000 specimens annually and performs rabies testing for the ...