这项工作得到了韩国科学和技术研究所项目 (No. 2E26190 和 2E26170) 和人类前沿科学项目 (RGY0089/2012) 的支持。

所描述的所有方法均已得到韩国科技研究院动物保育和使用委员会的批准. 
 1. 准备微驱动器和电极 <ol> <li> 组装微驱动器. 
	<ol> <li> 使用高分辨率3D 打印机打印微驱动器 (滑块、正文和外壳) 14 的各个部分。确保零件没有缺陷. </li>
		<li> 将滑块固定到带有螺钉的微驱动器体 (大小 000-120x1/4). </li>
		<li> 将螺母 (大小 000-120x5/64 十六进制) 焊接到螺钉的顶端. </li>
		<li> 顺时针旋转螺钉以将滑块向上或逆时针移动以将其向下移动. </li>
		<li> 用螺钉将外壳固定在机身上 (大小 000-120x1/4). </li>
	</ol> </li> <li> 在微驱动器上安装探测器。
	<ol> <li> 在双目显微镜下将微驱动器固定在水平位置. </li>
		<li> 使用带橡胶填料的扁平鳄鱼夹来抓取硅探针的挠性电缆, 同时小心地将探头从装运箱中分离出来, 用镊子。将鳄鱼夹子固定在机械手上. </li>
		<li> 使用机械手, 将硅探针放置在微驱动滑块上, 与移动方向平行. </li>
		<li> 应用一滴牙科水泥 (室温固化丙烯酸牙科修复材料) 将探头固定在滑块上。如果探头位置已移位, 请更正它。胶水是不推荐的, 因为它的治疗非常快, 使它很难重新调整电极的位置. </li>
		<li> 将探头连接器固定在带有牙科水泥的 C 型支架 (连接器支架) 上. </li>
	</ol> </li> <li> 清洗并将染料置于探头上. 
	<ol> <li> 修复探针清洗设备上的微/探头组合。该装置配备2小旋转泡沫海绵 (消毒拭子)。用机械手调节间隙. </li>
		<li> 用洗涤剂浸泡海绵. </li>
		<li> 慢慢地在海绵之间移动探头, 允许轻轻的触摸。在显微镜下监控清洗过程. </li>
		<li> 用蒸馏水冲洗探头。将探头浸入酒精中进行灭菌. </li>
		<li> 用泡沫拭子将 Dil (亲油性荧光染料) 涂抹在探针的柄的背面。这将允许在稍后阶段的可视化小腿位置在大脑中. </li>
	</ol> </li> <li> 组装 hat <ol> <li> 使用 3 d 打印机打印帽子 (头板、连接器和 cap) 14 的各个部分。3零件合在一起形成一个密封的外壳. </li>
		<li> 在磁头板的插槽中插入螺母 (大小 M2 和 00-90x1/4), 并用胶水和牙科水泥固定. </li>
		<li> 在瓶盖的插槽中插入垫圈并修复牙科水泥. </li>
	</ol> </li> </ol> 2。颅骨固定的手术方法 所有用于手术的工具都事先由灭菌消毒。干热珠装置是使用 forto 消毒工具, 成为污染, 并需要在手术中消毒. 
<ol> <li> 将异氟醚的水平设置为麻醉开始时的4%。将鼠标放在麻醉室中5分钟. </li>
	<li> 将鼠标安装在立体定向设备中. </li>
	<li> 将异氟烷水平降低到 1.5-2%。根据动物状态、呼吸速率和体温 20 调整手术的水平. </li>
	<li> 在眼睛上涂抹眼药膏. </li>
	<li> 用防腐剂 (iodopovidone) 将头皮剃光并清洁动物头部。在手术的所有步骤中保持无菌状态. </li>
	<li> 在头皮下注射布比卡因 (1 毫克/千克)。用细剪刀切开和移除小鼠头部的顶骨部分, 露出头骨的边缘。在手术中使用生理盐水和止血海绵清洗和控制出血. </li>
	<li> 使用刮刀工具拆卸骨膜. </li>
	<li> 查找头骨上的参考点 21 : bregma、λ、冠状和矢状缝合线。调整头部和 #39 的角度沿矢状轴, 使 Bregma 和 lambda 点处于同一高度. </li>
	<li> 在头骨上钻两个孔 (直径为0.5 毫米), 用于参考和接地电极。孔应该是大约1毫米尾鳍和1毫米侧向λ. </li>
	<li> 插入地面和参考电极 (尺寸 000-120x1/16 微型不锈钢螺丝线-耦合到针连接器). </li>
	<li> 在头骨上应用紫外线 (uv) 光合牙本质活化剂, 然后在45-60 秒应用 uv 光. </li>
	<li> 在头骨的边缘应用一层牙科水泥。避免在老鼠的皮肤和毛发上涂上水泥. </li>
	<li> 将磁头板固定在一个立体定位机械手上, 并将其置于颅骨上方。慢慢降低头部板, 直到它稍微触及头骨, 并应用牙科水泥在与头骨的交界处。让牙科水泥固化15分钟. </li>
	<li> 删除麻醉。将连接器和帽固定在头板上。在给 sub-cutaneous 注射 ketaprofen 5 毫克/千克后, 把老鼠放在笼子里. </li>
	<li> 给 sub-cutaneous 注射 ketaprofen 5 毫克/千克的两天, 并仔细监测任何疼痛的迹象。小鼠通常在20-40 分钟内从麻醉中苏醒。这种帽子的植入物相对较轻 (3.34g), 这样老鼠就不会有麻烦抬起头, 在迷宫中奔跑, 爬到他们家笼子的边缘. </li>
</ol> 3。行为训练 <ol> <li> 在后恢复期为7天后, 开始限制每天1毫升的水. </li>
	<li> 制作跑步机皮带, 切一块天鹅绒面料 (5 厘米乘1-2 米), 用热胶固定小物件。在皮带的边缘附加竖立的物体, 以免干扰鼠标的运动. </li>
	<li> 将两个端点缝合在一起, 固定在跑步机轮子上的皮带. </li>
	<li> 将磁头板的两个螺钉插入并拧紧到磁头固定板的插槽中, 将鼠标固定在跑步机上. </li>
	<li> 开始训练鼠标在跑步机上运行头部限制, 以获得水奖励。通过舔口提供水的奖励。最初, 把鼠标放在跑步机上10分钟, 每天3次. </li>
	<li> 当鼠标习惯于头部固定并开始移动皮带 (通常在3天后), 将训练会话持续时间增加到30分钟。2-3 周后, 一些老鼠在30分钟内跑了超过100次试验 (试验是皮带的完全旋转). </li>
	<li> 选择显示记录实验最佳行为表现的小鼠. </li>
</ol> 4。植入硅探针 <ol> <li> 将鼠标置于麻醉下. </li>
	<li> 通过固定头板将鼠标安装在立体定向装置中。用盐水清洗颅骨表面. </li>
	<li> 查找立体定向标记: bregma、λ、冠状和矢状缝合。测量到插入点的距离并标记它. </li>
	<li> 仔细地钻骨, 直到它变得薄而透明。在钻孔时用盐水滋润和清洁. </li>
	<li> 使用精确的力, 小心地去除薄骨和硬脑膜物质所有的时间保持脑表面湿润. </li>
	<li> 将微/硅探针组合固定在一个立体定位机械手上。把硅探针放在开颅上面将硅探针连接器固定在磁头板上. </li>
	<li> 连接记录放大器和接地/参考电极。用铝箔屏蔽鼠标以防止电磁噪声。启动记录系统以监视大脑的电活动. </li>
	<li> 使用微缓慢地将硅探针插入大脑。不断检查电信号, 机械手走过的距离, 和探针的小腿 (确保他们在大脑中穿透)。单位活动是可见的皮层, 而下方的白色物质是相对沉默。当小腿触及海马的锥体层时, 单位活动就会重现。从这一点上, 收回硅探针200和 #181; m (使用微驱动器的第二天, 使小腿回到金字塔层). </li>
	<li> 用骨蜡和矿物油的混合物覆盖大脑表面. </li>
	<li> 使用牙科水泥将微驱动器固定到磁头板上。让牙科水泥固化15分钟。然后将微驱动器从立体定向机械手中分离出来, 再戴上帽子帽. </li>
	<li> 将鼠标放回笼中, 并给出 ketaprofen 5 毫克/千克的 sub-cutaneous 注射。检查任何疼痛的迹象。这种手术比第一个更少的侵入和小鼠通常活跃在45分钟后, 他们从麻醉唤醒。然而, 当硅探针需要在大脑中稳定时, 让老鼠恢复一整天. </li>
</ol> 5。录制 <ol> <li> 在跑步机上安装鼠标头部固定。取下帽帽. </li>
	<li> 连接录音放大器并启动录制. </li>
	<li> 在第一天, 使用微驱动器将硅探针移动到海马的锥体层。每个逆时针旋转的螺丝移动小腿200和 #181; 我更深。在出现波纹振荡 13 和设备活动之前, 慢慢调整刀柄位置 (20-50 和 #181; m). </li>
	<li> 在第二天, 调整小腿位置和等待〜1小时之前记录海马活动, 而鼠标在皮带上运行。为了保持良好的记录信号质量几天, 删除硬物体和低天花板, 从鼠标和 #39; s 笼, 以尽量减少的机会, 帽子颠簸到坚硬的表面. </li>
</ol> 6。正在恢复探测器 <ol> <li> 将鼠标置于麻醉下. </li>
	<li> 通过固定头板将鼠标安装在立体定向装置中。取下帽帽. </li>
	<li> 将微驱动器上的立体定向机械手带上。将微驱动器固定在机械手上。从磁头板上拧下接头支架。卸下连接微型驱动器外壳和车身的螺钉. </li>
	<li> 在双目显微镜监视下, 用立体定向机械手缓慢拉动微驱动器/硅探针组合, 使壳体部分落后. </li>
	<li> 使用清洗设备清洗硅探针. </li>
</ol>

<ol>
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B., 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=Distinct+Contribution+of+Adult-Born+Hippocampal+Granule+Cells+to+Context+Encoding.">Distinct Contribution of Adult-Born Hippocampal Granule Cells to Context Encoding.</a> Neuron. 90 (1), 101-112 (2016).</li><li>Stark, E., Koos, T., Buzsaki, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diode+probes+for+spatiotemporal+optical+control+of+multiple+neurons+in+freely+moving+animals.">Diode probes for spatiotemporal optical control of multiple neurons in freely moving animals.</a> J Neurophysiol. 108 (1), 349-363 (2012).</li><li>Wu, F., 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=An+implantable+neural+probe+with+monolithically+integrated+dielectric+waveguide+and+recording+electrodes+for+optogenetics+applications.">An implantable neural probe with monolithically integrated dielectric waveguide and recording electrodes for optogenetics applications.</a> J Neural Eng. 10 (5), 056012 (2013).</li></ol>

作者没有什么可透露的。

慢性记录的神经元活动是关键的了解神经过程, 如海马位场。我们的方法来执行慢性硅探针 implantantation 区别自己从其他方法7,18,19,20的事实是, 它是相对简单的恢复电极封装在实验的结束。虽然地方细胞通常是在自由移动的条件下进行研究的, 但跑步机设备不仅能极大地简化实验设计和数据分析, 还可?...

硅探针为电生理记录提供了几个好处, 包括事实他们是设计的以锋利的外形最小化组织损伤, 并且他们提出一个密集地被包装的录音站点的一个精确布局1, 2、3、4。它们用于研究不同物种的各种系统, 包括人类3,5,6, 具有不同的方法1,7。然而, 由于它们的成本、脆弱性以及慢性实验的简便方法缺乏8, 它们的经常性使用仍然相对有限。在3D 印刷技术的最新进展使定制设计的设备, 如微型驱动器和头部板, 使更容易处理这些微妙的电极。在第一步中, 我们将描述如何构建和使用我们为植入慢性硅探针而开发的一套工具14。
虽然地方细胞通常是研究使用自由移动的动物在迷宫中运行, 最近他们也在虚拟环境中进行了15和在跑步机 apparatii9 (图 1A) 中进行了调查。这些实验方法提供的优势, 动物可以头部克制, 使使用2光子显微镜15, 补丁钳16, 和传感9,10,11技术更容易, 除了提供增强的控制动物行为和环境提示12。在第二步中, 我们将介绍训练小鼠和记录在跑步机设备中的细胞活动的程序。

<table><tbody><tr><td>Silicon Probe</td><td>Neuronexus</td><td>Buzsabi32</td><td>Recording electrode</td></tr><tr><td>Recording system</td><td>Intantech</td><td>RHD2132/RHD2000</td><td></td></tr><tr><td>3D printer</td><td>Asiga</td><td>Pico Plus 27</td><td>High resolution printer for micro-drive</td></tr><tr><td>3D printer</td><td>Stratasys</td><td>Mojo</td><td>Lower resolution printer for hat components</td></tr><tr><td>Stereotaxic apparatus</td><td>Kopf</td><td>Model 963</td><td></td></tr><tr><td>Binocular microscope</td><td>Leica</td><td>M60</td><td></td></tr><tr><td>Treadmill apparatus</td><td></td><td></td><td>We build them</td></tr></tbody></table>

implantation of chronic silicon probes and recording of hippocampal place cells in an enriched treadmill apparatus

了解大脑功能的一个重要条件是识别行为和细胞活性的相关性。硅探针是用于 large-scale 神经元活动的电生理记录的高级电极, 但其慢性植入的程序仍然不发达。已知海马位细胞的活动与动物在环境中的位置有关, 但其基本机制尚不清楚。为了研究地方细胞, 我们在这里描述了一系列的技术, 从制造设备的慢性硅探针植入到监测的地方现场活动在一个提示丰富的跑步机设备。一个微型驱动和帽子是由装配和紧固在一起 3 d 打印塑料零件。硅探针安装在微驱动器上, 清洗, 并涂上染料。第一次手术是为了修复老鼠头骨上的帽子。小地标是捏造的, 并附着在跑步机的皮带上。老鼠被训练在跑步机上运行头部固定。第二个手术是在海马内植入硅探针, 然后记录下宽带电生理信号。最后, 对硅探针进行恢复和清洗, 以便重新使用。分析的地方细胞活动在跑步机揭示了不同的地方领域的机制, 概述了该方法的好处。

一只老鼠被训练在两米长的皮带上运行, 没有提示 (图 1C)。在电极植入后, 一个新的相同长度的带, 但呈现3对线索安装在跑步机上, 为了生成 allocentric 空间表示12,14。宽带信号以3万赫兹的采样率记录, 使用250通道记录系统 (带有 USB 接口板和 custom-made Labview 接口的放大器板) 和两个硅探针 (图 1A...

Watch this Scientific Journal Video about Implantation of Chronic Silicon Probes and Recording of Hippocampal Place Cells in an Enriched Treadmill Apparatus at JoVE.com

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

我们描述了植入慢性硅探针的不同步骤, 并记录了老鼠体内的细胞, 这些小鼠在跑步头上固定在一个富含提示的跑步机上。

富氧运动器械中慢性硅探针的植入及海马位细胞的记录

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

implantation-chronic-silicon-probes-recording-hippocampal-place-cells

Research

JoVE Journal

Neuroscience

12.6K 次观看.  Korea Institute of Science and Technology. 我们描述了植入慢性硅探针的不同步骤, 并记录了老鼠体内的细胞, 这些小鼠在跑步头上固定在一个富含提示的跑步机上。

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

Recording Large-scale Neuronal Ensembles with Silicon Probes in the Anesthetized Rat

Large scale electrophysiological recordings from neuronal ensembles offer the opportunity to investigate how the brain orchestrates the wide variety of behaviors from the spiking activity of its neurons. One of the most effective methods to monitor spiking activity from a large number of neurons in multiple local neuronal circuits simultaneously is by using silicon electrode arrays1-3.
Action potentials produce large transmembrane voltage changes in the vicinity of cell somata. These output signals can be measured by placing a conductor in close proximity of a neuron. If there are many active (spiking) neurons in the vicinity of the tip, the electrode records combined signal from all of them, where contribution of a single neuron is weighted by its 'electrical distance'. Silicon probes are ideal recording electrodes to monitor multiple neurons because of a large number of recording sites (+64) and a small volume. Furthermore, multiple sites can be arranged over a distance of millimeters, thus allowing for the simultaneous recordings of neuronal activity in the various cortical layers or in multiple cortical columns (Fig. 1). Importantly, the geometrically precise distribution of the recording sites also allows for the determination of the spatial relationship of the isolated single neurons4. Here, we describe an acute, large-scale neuronal recording from the left and right forelimb somatosensory cortex simultaneously in an anesthetized rat with silicon probes (Fig. 2).

Extracellular recordings of neuronal activity using silicon probes in the anesthetized rat will be described. This technique allows information to be obtained across multiple brain areas from more than 100 neurons simultaneously. It provides information with single cell resolution about neuronal ensembles dynamics in multiple local circuits.

在麻醉大鼠的硅探头录制的大型神经元合唱

Large scale electrophysiological recordings from neuronal ensembles offer the opportunity to investigate how the brain orchestrates the wide variety of ...

科研

神经科学

Large-scale Recording of Neurons by Movable Silicon Probes in Behaving Rodents

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of neurons and circuits requires methods with the spatial selectivity and temporal resolution appropriate for mechanistic analysis of neural ensembles in the behaving animal, i.e. recording of representatively large samples of isolated single neurons. Ensemble monitoring of neuronal activity has progressed remarkably in the past decade in both small and large-brained animals, including human subjects1-11. Multiple-site recording with silicon-based devices are particularly effective because of their scalability, small volume and geometric design.
Here, we describe methods for recording multiple single neurons and local field potential in behaving rodents, using commercially available micro-machined silicon probes with custom-made accessory components. There are two basic options for interfacing silicon probes to preamplifiers: printed circuit boards and flexible cables. Probe supplying companies (<a href="http://www.neuronexustech.com/" >http://www.neuronexustech.com/</a>; <a href="http://www.sbmicrosystems.com/">http://www.sbmicrosystems.com/</a>; <a href="http://www.acreo.se/">http://www.acreo.se/</a>) usually provide the bonding service and deliver probes bonded to printed circuit boards or flexible cables. Here, we describe the implantation of a 4-shank, 32-site probe attached to flexible polyimide cable, and mounted on a movable microdrive. Each step of the probe preparation, microdrive construction and surgery is illustrated so that the end user can easily replicate the process.

We describe methods for large-scale recording of multiple single units and local field potential in behaving rodents with silicon probes. Drive fabrication, probe attachment to the drive and probe implantation processes are illustrated in sufficient details for easy replication.

移动硅探针的神经行为鼠害的大型记录

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of ...

Simultaneous Monitoring of Wireless Electrophysiology and Memory Behavioral Test as a Tool to Study Hippocampal Neurogenesis

Brainwaves amplitude obtained from electroencephalography (EEG) has been well-recognized as a basis for cognitive capacity, memory, and learning on animals and humans. Adult neurogenesis mechanism is also linked to memory and learning improvement. Traditionally, researchers used to assess learning and memory parameters in rodent models by behavioral tasks. Therefore, the simultaneous monitoring of behavioral changes and EEG is particularly interesting in correlating data between brain activity and task-related behaviors. However, most of the equipment required to perform both studies are either complex, expensive, or uses a wired setup network that hinders the natural animals&#8217; movement. In this study, EEG was recorded with a wireless electrophysiology device along with the execution of a novel object recognition task (NORT). The animal&#8217;s behavior was monitored simultaneously by a video tracking system. Both recordings were analyzed offline by their timestamps which were synched to link EEG signals with the animal&#8217;s actions. Subjects consist of adult Wistar rats after medium-term environmental enrichment treatment. Six skull screw electrodes were fixed in pairs on both hemispheres over frontal, central, and parietal regions and were referenced to an electrode located posterior of the nasal bone. NORT protocol consists of exposing the animal to two identical objects for 10 min. After 2 h and 24 h, one of the objects was replaced with a novel one. Exploration time for each object was monitored by a behavioral tracking software (BTS) and EEG data recording. The analysis of the EEG synced with behavioral data consists of estimations of alpha and beta relative band power and comparisons between novel object recognition versus familiar object exploration, between three experimental stages. In this manuscript, we have discussed electrodes manufacturing process, epidural electrodes implantation surgery, environmental enrichment protocol, NORT protocol, BTS setup, EEG &#8211; BTS coupling for simultaneous monitoring in real-time, and EEG data analysis based on automatic events detection.

The protocol presented here provides information on the simultaneous electroencephalography (EEG) and behavioral assessment in real-time. We have discussed all steps involved in this protocol as an attractive solution for researchers in many fields of neuroscience, particularly in learning and memory areas.

同时监测无线电生理学和记忆行为测试作为研究海马神经发生的工具

Brainwaves amplitude obtained from electroencephalography (EEG) has been well-recognized as a basis for cognitive capacity, memory, and learning on ...

行为学

Recording Spatially Restricted Oscillations in the Hippocampus of Behaving Mice

The local field potential (LFP) emerges from ion movements across neural membranes. Since the voltage recorded by LFP electrodes reflects the summed electrical field of a large volume of brain tissue, extracting information about local activity is challenging. Studying neuronal microcircuits, however, requires a reliable distinction between truly local events and volume-conducted signals originating in distant brain areas. Current source density (CSD) analysis offers a solution for this problem by providing information about current sinks and sources in the vicinity of the electrodes. In brain areas with laminar cytoarchitecture such as the hippocampus, one-dimensional CSD can be obtained by estimating the second spatial derivative of the LFP. Here, we describe a method to record multilaminar LFPs using linear silicon probes implanted into the dorsal hippocampus. CSD traces are computed along individual shanks of the probe. This protocol thus describes a procedure to resolve spatially restricted neuronal network oscillations in the hippocampus of freely moving mice.

This protocol describes the recording of local field potentials with multi-shank linear silicon probes. Conversion of the signals using current source density analysis allows the reconstruction of local electrical activity in the mouse hippocampus. With this technique, spatially restricted brain oscillations can be studied in freely moving mice.

行为小鼠海马空间受限振荡的记录

The local field potential (LFP) emerges from ion movements across neural membranes. Since the voltage recorded by LFP electrodes reflects the summed ...

Chronic Implantation of Multiple Flexible Polymer Electrode Arrays

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

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

多柔性聚合物电极阵列的慢性植入

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

Hybrid Microdrive System with Recoverable Opto-Silicon Probe and Tetrode for Dual-Site High Density Recording in Freely Moving Mice

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, conventional microdrive designs often only allow use of one type of electrode to record from single or multiple regions, limiting the yield of single-unit or depth profile recordings. It also often limits the ability to combine electrode recordings with optogenetic tools to target pathway and/or cell type specific activity. Presented here is a hybrid microdrive array for freely moving mice to optimize yield and a description of its fabrication and reuse of the microdrive array. The current design employs nine tetrodes and one opto-silicon probe implanted in two different brain areas simultaneously in freely moving mice. The tetrodes and the opto-silicon probe are independently adjustable along the dorsoventral axis in the brain to maximize the yield of unit and oscillatory activities. This microdrive array also incorporates a set-up for light, mediating optogenetic manipulation to investigate the regional- or cell type-specific responses and functions of long-range neural circuits. In addition, the opto-silicon probe can be safely recovered and reused after each experiment. Because the microdrive array consists of 3D-printed parts, the design of microdrives can be easily modified to accommodate various settings. First described is the design of the microdrive array and how to attach the optical fiber to a silicon probe for optogenetics experiments, followed by fabrication of the tetrode bundle and implantation of the array into a mouse brain. The recording of local field potentials and unit spiking combined with optogenetic stimulation also demonstrate feasibility of the microdrive array system in freely moving mice.

This protocol describes the construction of a hybrid microdrive array that allows implantation of nine independently adjustable tetrodes and one adjustable opto-silicon probe in two brain regions in freely moving mice. Also demonstrated is a method for safely recovering and reusing the opto-silicon probe for multiple purposes.

混合微驱系统,带可回收的光电探头和Tetrode,用于自由移动小鼠的双位点高密度记录

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, ...

用于记录幼年小鼠日常神经活动的 Micro-drive 植入物

A Lightweight Drive Implant for Chronic Tetrode Recordings in Juvenile Mice

In vivo electrophysiology provides unparalleled insight into the sub-second-level circuit dynamics of the intact brain and represents a method of particular importance for studying mouse models of human neuropsychiatric disorders. However, such methods often require large cranial implants, which cannot be used in mice at early developmental time points. As such, virtually no studies of in vivo physiology have been performed in freely behaving infant or juvenile mice, despite the fact that a better understanding of neurological development in this critical window would likely provide unique insights into age-dependent developmental disorders such as autism or schizophrenia. Here, a micro-drive design, surgical implantation procedure, and post-surgery recovery strategy are described that allow for chronic field and single-unit recordings from multiple brain regions simultaneously in mice as they age from postnatal day 20 (p20) to postnatal day 60 (p60) and beyond, a time window roughly corresponding to the human ages of 2 years old through to adulthood. The number of recording electrodes and final recording sites can be easily modified and expanded, thus allowing flexible experimental control of the in vivo monitoring of behavior- or disease-relevant brain regions across development.

作者聚焦：用于幼年小鼠慢性四极管记录的轻型驱动植入物  

Here, we describe a micro-drive design, surgical implantation procedure, and post-surgery recovery strategy that allow for chronic field and single-unit recordings from multiple brain regions simultaneously in juvenile and adolescent mice across a critical developmental window from postnatal day 20 (p20) to postnatal day 60 (p60) and beyond.

一种用于幼年小鼠慢性四极管记录的轻型驱动植入物

In vivo electrophysiology provides unparalleled insight into the sub-second-level circuit dynamics of the intact brain and represents a method of ...

开发用于狨猴精确神经记录的 x-y 电极台

Electrophysiology of Laminar Cortical Activity in the Common Marmoset

The marmoset monkey provides an ideal model for examining laminar cortical circuits due to its smooth cortical surface, which facilitates recordings with linear arrays. The marmoset has recently grown in popularity due to its similar neural functional organization to other primates and its technical advantages for recording and imaging. However, neurophysiology in this model poses some unique challenges due to the small size and lack of gyri as anatomical landmarks. Using custom-built micro-drives, researchers can manipulate linear array placement to sub-millimeter precision and reliably record at the same retinotopically targeted location across recording days. This protocol describes the step-by-step construction of the micro-drive positioning system and the neurophysiological recording technique with silicon linear electrode arrays. With precise control of electrode placement across recording sessions, researchers can easily traverse the cortex to identify areas of interest based on their retinotopic organization and the tuning properties of the recorded neurons. Further, using this laminar array electrode system, it is possible to apply a current source density analysis (CSD) to determine the recording depth of individual neurons. This protocol also demonstrates examples of laminar recordings, including spike waveforms isolated in Kilosort, which span multiple channels on the arrays.

作者聚焦:优化 Micro-Drive Systems 以实现 Marmoset 中的精确电极定位

Custom-built micro-drives enable the sub-millimeter targeting of cortical recording sites with linear silicon arrays.

普通狨猴层流皮层活动的电生理学

The marmoset monkey provides an ideal model for examining laminar cortical circuits due to its smooth cortical surface, which facilitates recordings with ...

富氧运动器械中慢性硅探针的植入及海马位细胞的记录

摘要

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此视频中的章节

在麻醉大鼠的硅探头录制的大型神经元合唱

移动硅探针的神经行为鼠害的大型记录

同时监测无线电生理学和记忆行为测试作为研究海马神经发生的工具

行为小鼠海马空间受限振荡的记录

多柔性聚合物电极阵列的慢性植入

混合微驱系统,带可回收的光电探头和Tetrode,用于自由移动小鼠的双位点高密度记录

一种用于幼年小鼠慢性四极管记录的轻型驱动植入物

一种用于幼年小鼠慢性四极管记录的轻型驱动植入物

一种用于幼年小鼠慢性四极管记录的轻型驱动植入物

普通狨猴层流皮层活动的电生理学