Name: construction of an improved multi-tetrode hyperdrive for large-scale neural recording in behaving rats
Uploaded: 2018-05-09T15:00:00.000+00:00
Duration: 10 min 4 s
Description: Watch this Scientific Journal Video about Construction of an Improved Multi-Tetrode Hyperdrive for Large-Scale Neural Recording in Behaving Rats at JoVE.com

我们感谢挪威科技大学 Kavli 神经系统科学研究所和神经计算中心莫石实验室为大鼠的慢性神经记录程序。这项工作得到了 NIH 赠款 R21 NS098146 和人类前沿科学计划的长期研究金 LT000211/2016-L 到 l 路。

1. 3D 机型的光固化成形
<ol><li>使用 stereolithographic 技术打印超空间引擎部件和配件。每个超空间引擎由十八梭、十八个梭螺栓和其他所有塑料件 (图 1) 组成。 注: 附件不是超空间引擎的一部分, 但对于超空间引擎的建设是必要的。</li>
</ol>2. 附件的准备工作 (图 2)。
<ol><li>
microdrive 机架的准备工作(图 2C)。<ol>
<li>在机架上用直径为0.71 毫米 (0.028 英寸) 钻头和直径为0.84 毫米 (0.033) 的钻头分别清洗并展开较小的通孔和较大的盲孔。</li>
		<li>将直径为0.89 毫米 (0.035 英寸) 的焊条切成17毫米长段, 两端圆, 并将每个导杆插入机架上的ø0.84 毫米 (0.033 英寸) 孔, 在外侧留11.5 毫米 (用螺纹棒冲洗)。</li>
		<li>完全插入六0-80 螺纹, 15.88 毫米 (5/8 英寸) 长扁头螺钉向下进入机架中的插槽。确保导杆和螺纹棒是直的和平行的。用稀释的牙科水泥填充插槽中的剩余空间。台式上的空气干燥15分钟。</li>
		<li>将焊接棒和螺钉粘附在带薄的超胶机架上, 使空气干燥15分钟。</li>
	</ol>
</li>
	<li>
核心站的准备工作(图 2E)。<ol>
<li>螺纹四孔与2-56 水龙头, 并使用 2-56, 4.76 毫米 (3/16) 长尼龙螺钉, 以确保核心在车站, 如果必要的话。</li>
	</ol>
</li>
	<li>
车削工具的准备(图 2F)。<ol>
<li>用4-40 丝锥将手柄上的孔螺纹。将加工尖端插入手柄中的插槽中, 并使用4-40、4.76 mm (3/16 英寸) 长杯螺钉固定。</li>
	</ol>
</li>
	<li>
超空间引擎持有者的准备(图 2G)。<ol>
<li>用8-32 丝锥将螺钉孔螺纹。使用 8-32, 9.52 毫米 (3/8) 长尼龙拇指螺钉, 以确保在使用时的超空间引擎。</li>
	</ol>
</li>
	<li>
杆定位复合体的准备(图 2H)。<ol>
<li>螺纹的茎从一侧与较大的孔 (顶部) 与8-32 丝锥的深度约7毫米. 螺纹较小的孔 (六在顶部, 十八在底部) 与0-80 水龙头。如果需要, 在顶部用直径为4.76 毫米 (3/16 英寸) 的钻头展开中心孔。</li>
		<li>将阀杆装配到顶部, 使用 8-32, 直径4.76 毫米 (3/16 英寸), 6.35 毫米 (1/4 英寸) 长肩螺钉。使用时, 用0-80、6.35 毫米 (1/4 英寸) 长螺钉将底部固定在顶部。</li>
	</ol>
</li>
</ol>3. 超空间引擎组件的准备工作 (图 3)。
<ol><li>超空间引擎螺母的准备(图 3A)。<ol>
<li>使用螺母支架 (图 2D), 螺纹螺母与 3/8-24 底触点直到平滑。</li>
	</ol></li>
	<li>超空间引擎内核的组件(图 3B)。<ol>
<li>用不同大小的钻头 (十二地线通孔 (内圈) 清洁和扩大芯孔: ø0.61 毫米 (0.024 英寸); 十八电子四极通孔 (中间圆环): ø0.66 毫米 (0.026) 首先, 然后ø0.71 毫米 (0.028); 十八导杆盲孔 (外环): ø0.84 毫米 (0.033 英寸))。</li>
		<li>螺纹的两个通孔上的核心和其余的八个盲孔 (四的一侧, 四附近的底部) 与0-80 水龙头。在盲孔上使用触底水龙头。</li>
		<li>使用 3/8-24 模具在核心底部创建外部线程。正确调整模具, 使超空间引擎螺母将适合新的线程。</li>
		<li>根据所需地线的数量, 将多个6毫米长段的23口径金属管 (套管) 插入核心的地线孔中, 如有必要, 将其胶合。将地线套管的两端锉至与芯的外冲, 并用直径为0.30 毫米 (0.012) 的钢丝清洁套管。</li>
		<li>完全插入十八 0-80, 15.88 毫米 (5/8 英寸) 长扁头螺钉头向下进入插槽中的核心。在此过程中, 请勿弯曲螺钉或损坏线程。</li>
		<li>使用杆定位复合体和核心站, 位置十八17毫米段的ø0.89 毫米 (0.035) 焊条在导杆孔的核心, 并锤击下来, 以与螺丝冲洗 (约5毫米)。</li>
		<li>如有必要, 纠正焊接棒和螺钉的位置, 然后拧紧中心肩螺钉和周围的六螺钉, 以确保杆在核心的向外方向。拧紧螺母上的核心 (与杆定位复杂), 并使核心进入超空间引擎的持有者, 以允许更容易地在立体镜下定位。</li>
		<li>用稀释的牙科水泥填充插槽, 将螺钉固定在芯上, 并允许空气干燥15分钟. 在牙科水泥变厚之前, 一次填充2-3 个插槽。在核心上刮掉多余的牙科水泥, 以保持与盾牌的适当配合。</li>
		<li>用薄的超级胶水将螺丝和棒粘到芯中, 使空气干燥15分钟。</li>
	</ol></li>
	<li>microdrive 的程序集(图 3C)。<ol>
<li>用钻头 (小孔: ø0.61 毫米 (0.024) 钻头; 较大的孔: ø0.89 毫米 (0.035 英寸) 钻头) 清洁和扩展航天飞机上的两个外孔。</li>
		<li>将梭螺栓插入螺栓座底座。注意方向。关闭螺栓支架盖, 紧紧抓住, 并通过0-80 水龙头在盖子孔慢慢通过。点击2-3 次, 直到平滑。</li>
		<li>用较小的开口将梭螺栓插入航天飞机一侧。将穿梭梭螺栓复合体倒在 microdrive 组装站基地。</li>
		<li>切割15毫米段的23口径金属油管和平滑两端, 然后定位油管在ø0.61 毫米 (0.024) 孔, 引导由插槽上的车站盖子。把套管锤进洞里, 直到上端与车站盖齐平。</li>
		<li>用磨砂轮移除套管上端的外半部。用直径为0.30 毫米 (0.012) 的金属丝清洗套管。用薄的超级胶水将套管粘到梭上, 确保不将梭螺栓粘在航天飞机上, 空气干燥15分钟。</li>
		<li>准备至少十八 microdrives, 测试 microdrive 机架上的 microdrive。确保航天飞机的螺栓能在航天飞机中平稳旋转, 并且整个 microdrive 沿着螺纹杆的长度自由移动。</li>
	</ol></li>
	<li>中央列的准备(图 3D)。<ol>
<li>将中心柱顶部和底部的沙子, 如果需要, 直到平坦。用0-80 分路器将中心柱上的两个孔螺纹。将0-80 个六角螺母 (3.18 毫米 (1/8 英寸) 宽, 1.19 毫米 (3/64 英寸) 高) 插入每个插槽中。</li>
	</ol></li>
	<li>超空间引擎盖的准备工作(图 3E)。<ol>
<li>使用非磁性镊子, 将四磁铁 (直径3毫米, 1 毫米厚) 粘附到四井中, 将其与电极接口板上的 N 和 S 极相匹配。</li>
	</ol></li>
	<li>指南套管到捆绑包中的程序集(图 3F)。<ol>
<li>放置十八30规, 薄壁套管 (ID 0.19 毫米, 0.0075 英寸) 到ø2.29 毫米 (0.09 英寸) 热收缩管 (3-5 毫米长, 间隔沿捆绑由5-10 毫米)。使所有套管在捆绑的一端彼此刷新。</li>
		<li>使用热枪收缩热收缩管, 直到束紧。轻轻地挤压包裹, 按需要的形状 (圆形或椭圆形)。确认所有套管都处于正确的位置, 没有扭曲、交叉或弯曲。</li>
		<li>标记在套管上焊接的区域。发起部分应长度为26毫米, 而焊接部分应为5-10 毫米. 将收缩管移至焊接标记以防止扩散。</li>
		<li>在旋转捆绑包时, 将焊剂应用于一个焊锡区和焊料。室温下冷却至少1分钟. 重复此步骤, 将同一区域再焊两次。通过焊接, 不应用焊剂和填料材料, 平滑焊接部分。室温下冷却至少1分钟。</li>
		<li>用金刚石砂轮以最高的速度切开捆绑到适当的长度, 擦亮两端调整长度 (发起部分:26 毫米, 被焊接的部分: 5-10 毫米根据需要)。用直径为0.18 毫米 (0.007) 的金属线在立体镜下清洁导轨套管。</li>
	</ol></li>
	<li>准备 tetrodes。类似的过程被描述为8、16、17 。<ol>
<li>调整水平 T 杆的高度和磁力搅拌器的位置, 使 t 杆的横臂位于磁搅拌器中心的正上方。将 S 钩的一端钩到一个小的磁力搅拌杆的中心, 然后将它们粘在一起。用压缩空气和乙醇湿巾清洁电子四极制造空间。</li>
		<li>将一块单电子四极线的两端绕在一起40厘米长, 然后用一块铜胶带固定。</li>
		<li>握住铜带, 提起钢丝圈。将末端与铜带相反放置在 T 条的水平臂上。轻轻地降低铜带 (而另一端仍在 t 栏上), 扭转一次, 并将铜胶带放到 t 栏上。电子四极圆现在在图八 ("∞") 配置与铜磁带坐在横杠的十字架之上。</li>
		<li>用一只手轻轻握住 T 条上的铜胶带。用另一只手钩住 s 钩的自由端 (用磁力搅拌连接到另一端) 通过电子四极线圈的底部, 轻轻松开 s 钩, 让它通过 s 钩的重量理顺四导线。</li>
		<li>调整水平杆的高度, 直到 S 钩的底部约1厘米以上的磁性搅拌器板的中心。</li>
		<li>向下弯曲铜胶带的边缘以将其固定在水平条上。检查四直电子四极线的眼睛, 然后清除任何碎片。</li>
		<li>打开搅拌器扭四导线在速度大约60转每分钟, 直到角度在二对面解开导线之间是关于。</li>
		<li>将热枪设置为210摄氏度, 并加热扭曲的电线, 沿直线长度的电线从不同的角度, 以2分钟, 以熔化的 VG 债券涂层。</li>
		<li>轻轻地提起 S 钩, 用细剪刀剪下电子四极的下端。</li>
		<li>用手指把横杠上的铜胶带用剪刀把铜胶带两边的导线剪掉, 然后取下铜胶带。切断水平条上的剩余导线, 释放电子四极。</li>
		<li>将已完成的电子四极放在无尘箱中存放。准备至少二十五 tetrodes。</li>
	</ol></li>
</ol>4. 超空间引擎的装配 (图 4)。
<ol><li>将指南套管插入超空间引擎核心 (图 4A)。<ol>
<li>卸下热收缩管并将4毫米的硅管段 (ID 1.02 毫米 (0.04))、外径2.16 毫米 (0.085 英寸)) 沿捆绑到焊接/发起边框上。在超空间超空间中楔入缝隙以加宽中心孔, 允许间隔在硅管周围滑动。当垫片位于硅管中心时, 取下楔形。</li>
		<li>通过将直径 0.18mm (0.007) 金属导线的长段 (10 厘米) 通过每个套管放置到超空间超空间中的一个特定电子四极孔中, 以防止该过程中的导线或套管的任何交叉, 从而将指南套管的位置组织在捆绑包中。弯曲两端的电线, 以保持他们到位。</li>
		<li>把套管通过各自的孔在核心, 小心避免弯曲或跨越他们之间, 直到每个套管的自由端是至少2毫米外的电子四极孔的上端。把螺母拧到芯上, 小心防止垫片旋转, 以确保垫片的安全。将一滴非常稀的牙科水泥从核心的顶端放到套管的交界处, 以确保它们的相对位置。</li>
		<li>从包的焊接端剪下导丝, 从自由端缩回, 从套管中取出。</li>
	</ol>
</li>
	<li>将 microdrives 装配到超空间引擎核心 (图 4B)。在超空间引擎中, microdrives 的详细空域安排以前被描述为11,13。<ol>
<li>将 microdrives 慢慢地和仔细地加载到核心的每个螺纹杆上。确认 (1) 23 口径 microdrive 套管顺利进入电子四极孔, (2) 30 口径导套管顺利进入23口径 microdrive 套管, (3) 梭螺栓沿螺纹杆平稳转动。把 microdrives 拧到 1.0-1.5 毫米以上的螺纹棒的下端。</li>
		<li>切割十八片聚酰亚胺油管 (ID 0.11 毫米 (0.0045), 外径0.14 毫米 (0.0055 英寸)) 成38-43 毫米段 (导套管束长度加7毫米)。用直径为0.08 毫米 (0.003) 的钢丝清洗每管。</li>
		<li>反转核心, 把聚酰亚胺管小心地插入到指南套管从焊接端, 并推动他们一路在立体镜下。将核心垂直翻转, 将聚酰亚胺管的上端粘附在 microdrive 套管上, 用厚的超级胶水。将核心倒置, 让胶水干15分钟。</li>
		<li>在上端切下多余的聚酰亚胺油管, 在 microdrive 套管外留下 0.5-1.0 毫米。</li>
	</ol>
</li>
	<li>
接地线的装配(图 4C)。<ol>
<li>从涂层钢丝 (直径为0.20 毫米 (0.008 英寸)、裸直径0.13 毫米 (0.005 英寸)) 中切割所需长度为25-30 毫米的地线的数量。从导线的两个尖端剥离2毫米的塑料绝缘, 并将每个端插入到 6-8 mm 长30口径套管的两端。压扁套管的两端, 以确保连接到各自的导线上。</li>
		<li>使用 Dremel 工具将套管切成两半, 以便从每条线上创建二个完整的地线。</li>
		<li>将30口径套管的圆端插入核心的地线套管的上端, 并按下以使插入紧密。</li>
	</ol>
</li>
	<li>
电极接口板的装配(图 4D)。<ol>
<li>将中心柱插入核心, 并用两个0-80、7.94 毫米 (5/16 英寸) 长的套筒头螺钉固定。胶水如果需要, 使中心柱稳定的核心。</li>
		<li>展开 EIB-72-QC-Large 板中插槽的部分, 该槽对应于中心柱上的两个丝锥孔, 直径为1.2 毫米。将电极接口板连接到中心柱上, 有两个0-80、3.97 毫米 (5/32 英寸) 长盘头螺钉。请确保主板位于中心, 并且安全。</li>
	</ol>
</li>
	<li>
连接接地线(图 4E)。<ol>
<li>在中心柱周围布线每根地线, 并将暴露的自由端连接到电极接口板上, 并在指定的接地孔上使用金针。</li>
	</ol>
</li>
	<li>
将tetrodes 加载到超空间引擎中, 如前所述16,17 。<ol>
<li>将每个电子四极小心地装入 microdrives 的聚酰亚胺管中, 小心不要在过程中弯曲它们。</li>
		<li>将自由端线轻轻地送进电极接口板中的指定孔中, 并用金针电连接。</li>
		<li>将 tetrodes 单独剪切到适当的长度。确认在切割后 tetrodes 的部分从聚酰亚胺管的下端凸出是直的, 否则取代整个电子四极和裁。</li>
	</ol>
</li>
	<li>
附上盾牌。
	<ol>
<li>使用四 0-80, 3.97 毫米 (5/32 英寸) 的盘头螺钉将防护罩连接到核心。盾上的数字应该与电极接口板上的数字相匹配。</li>
	</ol>
</li>
	<li>
电镀电子四极提示。
	<ol>
<li>使用装有 ADPT-NZ-EIB-36 连接器和 ADPT-EIB-72-QC-HS-36 适配器17的 NanoZ 电镀设备 tetrodes 的提示。或者, 按照其他地方16中的说明, 手动将它们板成一个。使用前的电子四极提示(例如, 植入前一天), 因为阻抗将随着时间的推移逐渐增加电镀。更换在电镀过程中短路或受阻的 tetrodes, 将其剪成适当的长度, 再重新制版。</li>
	</ol>
</li>
	<li>
完成超空间引擎的最终完成(图 4F)。<ol>
<li>将 tetrodes 粘附到其聚酰亚胺管上, 如前所述16。收回所有他们回到他们的指南套管, 所以镀的提示不暴露。</li>
		<li>螺丝四 0-80, 6.35 毫米 (1/4 英寸) 长套筒头螺钉进入四孔附近的超空间引擎核心。</li>
		<li>使用立体镜, 慢慢降低每个电子四极, 直到电子四极尖端正上方的导向套管的边缘。同时, 在导套管束中定位每个电子四极的位置。电子四极的位置图是重建记录站点的关键。</li>
		<li>将瓶盖连接到驱动器上, 并将超空间引擎正确存储在植入中。</li>
	</ol>
</li>
</ol>

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作者没有什么可透露的。

在这里, 我们描述了建设一个新开发的超空间引擎由十八独立移动 tetrodes 组成的过程。该驱动器可以通过在许多可用的硬件商店购买的负担得起的部件构建, 并与 stereolithographic 打印创建的组件结合使用。超空间超光速可以用标准的手术方法长期植入大鼠的头骨, 并且能够记录细胞外神经活动, 而动物则执行各种行为任务。
超空间引擎保留了原始诺顿超空间引擎的许多可取特性, 包括从驱动器中心13向外定向的三脚架 microdrives, 它为 tetrodes 提供了可靠的支持。一旦植入, 超空间引擎提供了一个清醒的动物的大脑内的 tetrodes 小运动的执行, 具有相当的精确度。一个在螺纹杆上的穿梭机的全轮对应于317.5 µm 的线性位移。通过适当的训练, 实验者可以在1/16 个回合步骤 (20 µm) 中推进航天飞机。我们设计的超空间引擎用于成年大鼠, 但驱动器可以很容易地用于任何动物的身体大小350克或更大 (受头部大小限制)。该装置的一个限制可能在记录的限制深度, 因为 tetrodes 沿螺纹棒的最大行程距离约7毫米, 这可能会落后于一些动物的大脑更深的结构。
Stereolithographic 打印提供了足够的分辨率以高保真度非常详细地创建塑料组件, 并且以前曾用于超空间引擎制造12,19,20。在这种情况下, 使用了通常通过第三方生产设施提供的工业打印机。在那里, 所有的超空间引擎部件都是精确打印的, 包括超空间超空间的核心, 尽管其复杂的几何形状, 以及小的结构, 如ø直径为0.6 毫米的孔和0.3 毫米薄墙。这种精度使光固化成形成为制造超空间引擎组件的理想选择。根据以前的经验, 成本较低的台式机3D 打印机不太可能具有精确复制所需的超空间引擎组件所需的精度。尽管如此, stereolithographic 技术还是有其局限性。首先, 它的材料选择有限。我们选择的超空间引擎的塑料是最耐用的, 我们已经测试, 但它仍然不是最佳的制造非常小的件。航天飞机和梭螺栓需要格外小心地处理, 因为它们在准备过程中会断裂。由于材料的热偏转温度大约50°c, 塑料组分不耐高压。另外, 所用的印花材料不耐丙酮。这些问题可以解决时, 新的光固化成形材料的开发和测试。然而, 考虑到成本相对较低的光固化成形, 技术和成本的优势远远超过了缺陷。其次, 由于光固化成形的性质, 在该过程中, photopolymers 由 uv 激光器光化学法固化, 形成所需的3D 模型21的单个层, stereolithographic 打印所创建的对象容易受到 UV 光的影响。因此, 将它们暴露在强紫外线下 (例如, 阳光直射) 可以不可逆转地减少其体力 (根据与印刷车间的个人通信)。考虑到实验室空间中的环境紫外线 (例如, 从荧光灯中), 最好在不使用的情况下将 stereolithographic 组件存储在一个黑暗的盒子里, 这将会保留组件的物理强度。此外, 除了紫外线外, 还必须使用其他方法在手术前对超空间超光速进行消毒。这个测试超光速引擎仍然在一个正常的实验室环境下, 在四月内被植入的大鼠, 没有任何迹象表明体力或性能下降。
这个超空间引擎的3D 可打印性也允许快速修改和灵活的重新设计。例如, 超空间引擎可以很容易地被修改为目标分离的多脑区域11。此外, 这个驱动器可以调整, 以允许同时监测神经活动和局部大脑操作。将微透析探针与 tetrodes 阵列结合在一起, 可以在神经记录22期间通过各种药物的注入, 使神经元的药理活化和失活。此外, 通过将光纤纳入电子四极束和 optogenetic技术19 中, 可以激活或停用用于表达光敏通道的神经元。此外, 该驱动器可以很容易地 rescaled 与较少数量的 tetrodes 的动物与较小的头部大小, 如小鼠或幼鼠。
总之, 容易的易变性加上更简单, 更实惠的方法构建一个有效的神经记录植入物, 可以可靠和准确地再现, 使这个超空间引擎成为一个强大的工具, 在这一领域。

在系统神经科学领域, 科学家研究了神经相关的认知过程, 如空间导航, 记忆和决策。对于这些类型的研究, 在动物行为过程中监测许多单个神经元的活动是至关重要的。在过去的几十年中, 已经取得了两个重要进展, 以满足小动物细胞外神经记录的实验需要1,2,3。首先是开发的电子四极, 一捆四微丝用于记录神经元的神经活动同时1,2,4。电子四极四通道的活动的差分信号振幅允许从许多同时记录的单元格5 中分离单个神经元活动。此外, 微丝的柔韧性使得电子四极的稳定性能够最大限度地减少电子四极与目标细胞种群之间的相对位移。Tetrodes 现在被广泛使用, 而不是一个单一的电极为许多脑研究在不同的物种, 包括啮齿目动物1,2,6, 灵长类7, 昆虫8。其次是开发一个超空间引擎运载多个独立移动的 tetrodes, 它允许同时监测神经活动从大群神经元从多个记录位置3, 9,10,11,12。
为小动物提供可靠和负担得起的多电子四极记录装置是有限的。经典的超空间引擎最初由布鲁斯诺顿13开发, 已成功用于在过去两年中在许多实验室中自由行为的老鼠的神经记录9,10,14, 15. 但是, 由于技术原因, 构建诺顿驱动器所需的原始组件现在很难获得, 并且与最近改进的数据获取接口不兼容。另一个很好接受的超空间引擎设计要求 microdrives 单独手工制作, 这可能产生不一致的结果并消耗大量的时间12。为了记录大鼠不同脑区的神经活动, 我们利用 stereolithographic 技术开发了一种新的超光速引擎。我们寻求满足以下要求: (1) 新的超空间引擎必须允许精确的 tetrodes 在大脑中的位移, 并提供来自多个目标区域的稳定记录;(2) 新的超空间引擎必须与最近开发的磁 quickclip 系统兼容, 以便于方便连接;(3) 新的超光速引擎可以用容易获得的材料准确地再现。在这里, 我们提供了一个技术, 建立3维可打印的超空间引擎, 包含十八独立移动 tetrodes, 基于诺顿设计。在该协议中, 我们描述了新的超空间引擎的制造过程的细节, 我们已经成功地记录了 postrhinal 和内侧嗅皮层的单神经元动作电位和局部场电位, 在一周内自由地在自然觅食任务中的行为鼠。

<table><tbody><tr><td>Welding rod</td><td>Blue Demon</td><td>ER308L-035-01T</td><td>Stainless steel, 0.035&quot; in diameter</td></tr><tr><td>Screw</td><td>McMaster</td><td>91771A060</td><td>Stainless steel, flat head, 0-80&amp;nbsp;thread,&amp;nbsp;5/8&quot;&amp;nbsp;in length</td></tr><tr><td>Screw</td><td>McMaster</td><td>91772A051</td><td>Stainless steel, pan head, 0-80&amp;nbsp;thread,&amp;nbsp;5/32&quot;&amp;nbsp;in length</td></tr><tr><td>Screw</td><td>McMaster</td><td>92196A056</td><td>Stainless steel, socket head, 0-80&amp;nbsp;thread,&amp;nbsp;5/16&quot;&amp;nbsp;in length</td></tr><tr><td>Screw</td><td>McMaster</td><td>92196A055</td><td>Stainless steel, socket head, 0-80&amp;nbsp;thread,&amp;nbsp;1/4&quot;&amp;nbsp;in length</td></tr><tr><td>Screw</td><td>McMaster</td><td>95868A131</td><td>Nylon,&amp;nbsp; socket head, 2-56&amp;nbsp;thread,&amp;nbsp;3/16&quot;&amp;nbsp;in length, black</td></tr><tr><td>Screw nut</td><td>McMaster</td><td>90730A001</td><td>Stainless steel, narrow hex,&amp;nbsp; 0-80&amp;nbsp;thread</td></tr><tr><td>Shoulder screw</td><td>McMaster</td><td>90298A213</td><td>Stainless steel, 8-32 thread, 3/16&quot; in diameter, 1/4&quot; in length</td></tr><tr><td>Cup screw</td><td>McMaster</td><td>92313A105</td><td>Stainless steel, 4-40 thread, 3/16&quot; in length</td></tr><tr><td>Thumb screw</td><td>McMaster</td><td>94323A592</td><td>Nylon, 8-32&amp;nbsp;thread,&amp;nbsp;3/8&quot;&amp;nbsp;in length, black</td></tr><tr><td>Magnet</td><td>Apex</td><td>M3X1MMDI</td><td>Neodymium, 3 mm X 1 mm disc</td></tr><tr><td>Metal tubing</td><td>Small Parts</td><td>B00137QHNS</td><td>Stainless steel, 23 gauge, 0.0253&quot; OD, 0.013&quot; ID, 0.006&quot; wall</td></tr><tr><td>Metal tubing</td><td>New England Small Tube</td><td>Custom-made</td><td>Stainless steel, 30 gauge, 0.012/0.0125&quot; OD, 0.007/0.008&quot; ID, full hard</td></tr><tr><td>Heat-shrink tubing</td><td>McMaster</td><td>7856K72</td><td>0.09&quot; ID before shrinking, blue</td></tr><tr><td>Silicone tubing</td><td>A-M Systems</td><td>807300</td><td>0.040&quot; ID, 0.085&quot; OD</td></tr><tr><td>Polyimide tubing</td><td>A-M Systems</td><td>823400</td><td>0.0045&quot; ID, 0.0005&quot; wall</td></tr><tr><td>Ground wire</td><td>A-M Systems</td><td>791500</td><td>0.005&quot; bare, 0.008&quot; coated, half hard</td></tr><tr><td>Tetrode wire</td><td>California Fine Wire</td><td>Custom-made</td><td>0.0007&quot; in diameter, platinum-iridium (90%-10%), HML and VG coating</td></tr><tr><td>EIB</td><td>Neuralynx</td><td></td><td>EIB-72-QC-Large</td></tr><tr><td>Gold pins</td><td>Neuralynx</td><td></td><td>large EIB pins</td></tr><tr><td>Tap</td><td>Balax</td><td>01302-000</td><td>M1.2 thread size</td></tr><tr><td>Tap</td><td>McMaster</td><td>2522A811</td><td>0-80 thread size, bottoming</td></tr><tr><td>Tap</td><td>McMaster</td><td>2522A771</td><td>0-80 thread size, plug</td></tr><tr><td>Tap</td><td>McMaster</td><td>26955A94</td><td>3/8&quot;-24&amp;nbsp;thread size, bottoming</td></tr><tr><td>Tap</td><td>McMaster</td><td>2522A713</td><td>2-56&amp;nbsp;thread size</td></tr><tr><td>Tap</td><td>McMaster</td><td>2522A715</td><td>4-40&amp;nbsp;thread size</td></tr><tr><td>Tap</td><td>McMaster</td><td>2522A718</td><td>8-32&amp;nbsp;thread size</td></tr><tr><td>Die</td><td>McMaster</td><td>2576A457</td><td>3/8&quot;-24&amp;nbsp;thread size, 1&quot; OD</td></tr><tr><td>Drill bit</td><td>McMaster</td><td>30585A82</td><td>Wire gauge&amp;nbsp;65,&amp;nbsp;0.035&quot; in diameter</td></tr><tr><td>Drill bit</td><td>McMaster</td><td>30585A83</td><td>Wire gauge&amp;nbsp;66,&amp;nbsp;0.033&quot; in diameter</td></tr><tr><td>Drill bit</td><td>McMaster</td><td>30585A87</td><td>Wire gauge&amp;nbsp;70,&amp;nbsp;0.028&quot; in diameter</td></tr><tr><td>Drill bit</td><td>McMaster</td><td>30585A88</td><td>Wire gauge&amp;nbsp;71,&amp;nbsp;0.026&quot; in diameter</td></tr><tr><td>Drill bit</td><td>McMaster</td><td>30585A91</td><td>Wire gauge&amp;nbsp;73,&amp;nbsp;0.024&quot; in diameter</td></tr><tr><td>Drill bit</td><td>McMaster</td><td>8870A23</td><td>3/16&quot;&amp;nbsp;in diameter</td></tr><tr><td>Dremel disc</td><td>Wagner</td><td>31M</td><td>Diamond coated, 22 mm in diameter, 0.17 mm in thickness</td></tr><tr><td>Steel wire</td><td>Precision Brand</td><td>21212</td><td>0.012&quot; in diameter, full hard</td></tr><tr><td>Steel wire</td><td>Precision Brand</td><td>21007</td><td>0.007&quot; in diameter, full hard</td></tr><tr><td>Steel wire</td><td>A-M Systems</td><td>792700</td><td>0.003&quot; in diameter, half hard</td></tr><tr><td>Super glue</td><td>Loctite</td><td>LT-40640</td><td># 406</td></tr><tr><td>Super glue</td><td>Loctite</td><td>LT-41550</td><td># 415</td></tr><tr><td>Dental acrylic powder&amp;nbsp;</td><td>Teets</td><td>223-3773</td><td>Coral</td></tr><tr><td>Dental acrylic liquid</td><td>Teets</td><td>223-4003</td><td></td></tr></tbody></table>

construction of an improved multi-tetrode hyperdrive for large-scale neural recording in behaving rats

在清醒的动物中监测大量神经元的活动模式, 是系统神经科学领域的一项宝贵技术。该技术的一个关键组成部分是将多个电极精确放置到所需的脑区, 并维持其稳定性。在这里, 我们描述了一个构建3维可打印超空间引擎的协议, 其中包括十八个独立调整的 tetrodes, 专门针对在自由行为的大鼠体内的活体细胞外神经记录。附着在 microdrives 上的 tetrodes 可以单独推进到轨道上的多个脑区, 也可以用来将电极阵列放置到较小的区域。多重 tetrodes 允许同时检查许多单个神经元的动作电位, 以及在活动行为期间大脑神经元的局部磁场电位。此外, 该设计提供了更简单的3D 绘图软件, 可以很容易地修改不同的实验需要。

我们用一个新的超光速引擎来获得试验结果。该驱动器配备了 tetrodes 从ø17µm (0.0007), 聚酰亚胺涂层铂铱 (90%-10%) 线。tetrodes 的尖端被镀在铂黑溶液中, 以减少电极阻抗到100和 200 kΩ在1赫之间。超空间超空间被植入4.6 毫米左侧的中线和0.5 毫米前的横窦的头骨上的550克, 雄性长埃文斯鼠。另外的地线连接在小脑上的颅骨螺钉上。所有程序都是由贝勒医学院的机构动物护理和使用委员会 (IACUC) 批准的, 与以前描述的18类似。手术后立即植入, tetrodes 1 毫米进入大脑。随后几天, 使用了不超过80µm 的较小的高级增量。tetrodes 被允许稳定在每个推进以后至少20小时在神经录音被执行之前。
为了记录神经活动, 超空间引擎连接到一个 headstage 前置放大器 (Neuralynx, HS-72-QC), 后者被连接到一个数据采集系统与可编程放大器 (Neuralynx, 数字山猫 SX)。局部磁场电位被引用到地线, 采样在2赫, 带通滤波在 0.1-500 赫兹. 单位活动被引用到一个电子四极, 没有任何可观察的活动, 从大脑表面的500µm, 采样在32赫, 带通滤波在600 Hz-6 赫。仅记录了阈值为50µV 的峰值波形。
图 5A说明了从位于 postrhinal 皮层 (2.1 毫米以下脑表面) 的电子四极中记录的神经活动, 而该动物在植入后三周内可以自由觅食1.5 米开放盒内。记录会话持续了大约30分钟, 记录的单元在整个会话中保持稳定 (由穗形波形的小变化证明)。图 5B显示了从四个不同的 tetrodes 位于内侧嗅皮层 (3.4-3.7 毫米深) 同时记录的局部场电位, 而同一动物在植入后七周积极探索开放竞技场。在θ频率范围 (6-10 Hz) 中存在明显的场电位活动。通过分类软件 MClust (红褐色) 分离出单个神经元尖峰数据, 并通过自定义编写的 Matlab 脚本对局部场电位数据进行可视化。以前17显示了低质量电子四极录制的示例, 可能是由于准备不足的驱动器导致的。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/57388/57388fig1.jpg"> 图 1: 由 stereolithographic 技术创建的超空间引擎组件.图像的 3 d 可打印的超空间引擎组件 (1¢硬币的大小比较)。(A) 超空间引擎核心;(B) 防护盾;(C) 保护帽;(D) 中央列;(E) 螺母;(F) 间隔;(G) 航天飞机;(H) 航天飞机螺栓。刻度条: 1 厘米。这些组件是由 UnionTech RSPro450 打印机创建的, 使用塑料材料 Somos 演化128。<a href="/files/ftp_upload/57388/57388fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/57388/57388fig2.jpg"> 图 2: 用于超空间引擎结构的定制设计附件.这些附件专门设计用于帮助准备超空间引擎。它们的主要组件是通过 stereolithographic 打印创建的。(A) 梭螺栓固定架, 它在攻丝时确保梭螺栓的固定。(B) microdrive 装配站, 它引导套管插入航天飞机。(C) microdrive 机架, 它有助于测试装配的 microdrives, 并在胶合套管时将其保存到位。1: microdrive 机架底座;2: microdrive 机架, 螺钉完全插入插槽中;3: microdrive 机架准备使用。(D) 螺母保持器, 它在穿线孔时持有超空间超光速螺母。(E) 超空间引擎核心站, 它在对导杆进行锤击时保证核心。(F) 车削工具, 驱动梭螺栓在航天飞机中旋转。(G) 超空间引擎架, 它有助于将超光速引擎置于立体镜下。持有人也保护 tetrodes 后, 他们已经加载到超空间引擎。(H) 杆定位复合体, 它有助于在超空间引擎核心中定位螺纹杆和导杆。1: 复杂的主要组分;2: 复杂在装配以后的上部部分;3: 在使用中的杆定位复合体。<a href="/files/ftp_upload/57388/57388fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/57388/57388fig3.jpg"> 图 3: 装配前的超空间引擎部件的准备.图像显示了超空间超空间的核心和 microdrive 的准备过程, 以及其他准备好的超空间引擎部件。(A) 螺纹超空间超光速螺母。(B) 准备超空间引擎核心。1: 核心与为坚果创造的外在螺纹;2: 核心放在核心站内, 螺钉完全插入插槽中;3: 导向杆定位由杆位置复合, 准备将锤击成芯;4: 用稀牙水泥填补槽内剩余空间;5: 准备好的超空间引擎核心的上部部分。(C) microdrive 的准备工作。1: 一个梭螺栓放置在梭螺栓支架底座上, 注意较小的开口朝向远离实验者;2: 穿线梭螺栓内的螺纹;3: 插入梭螺栓入梭;4: microdrive 放置在 microdrive 的装配站底座上, 由站盖引导的套管, 准备插入;5: microdrive 与上部套管末端的外部一半去除 (由箭头指示);6: 在 microdrive 机架上测试组装 microdrives。(D) 具有螺纹孔和插入螺钉螺母的中心柱。(E) 一个超空间引擎盖, 四磁铁粘在水井中。(F) 一个36毫米长的引导套管束, 在左侧焊接部分。<a href="/files/ftp_upload/57388/57388fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/57388/57388fig4.jpg"> 图 4: 超空间引擎的组件.显示超空间引擎组件阶段的图像。(A) 将指南套管插入核心。1: 导套管束滑入硅管和垫片;2: 一个指南套管被安置在它的指定的孔在核心。手工书写显示指南套管的组织;3: 引导套管推入核心;4: 核心与指南套管插入和由坚果巩固。(B) microdrives 到核心的程序集。1: 核心与 microdrives 装载;2: microdrives 与聚酰亚胺管子插入在套管。(C) 将接地导线插入核心。(D) 电极接口板的附件。1 : 超空间引擎与入的中央专栏 ;2: 超光速引擎与电极接口板连接到中央专栏。(E) 接地线与电极接口板上指定孔的连接。(F) 已完成的超空间引擎准备植入 (总重量为20克)。<a href="/files/ftp_upload/57388/57388fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/57388/57388fig5.jpg"> 图 5: 超空间引擎记录的神经信号.有代表性的记录显示在行为大鼠大脑中的单位神经活动和局部磁场电位。(A) 二维聚类图, 用于说明位于 postrhinal 皮层 (深度: 2.1 毫米) 的电子四极同时记录的神经元的峰值。左: 散点图显示从电子四极的两个电极记录的峰值峰峰值振幅之间的关系。每个点对应一个穗。尖峰簇可能来自同一个细胞。四簇是彩色编码的。刻度条:20 µV。右: 尖峰波形 (表示左侧显示的颜色编码单元格的 S.D.). 注意波形的小变化。缩放栏: 200 µs. (B) 当大鼠自由觅食时, 从四个不同的 tetrodes 位于内侧嗅皮层 (深度: 3.4-3.7 毫米) 同时记录的θ频率范围内的局部磁场电位的痕迹。左下角刻度条: 500 µV;右下角缩放栏: 100 毫秒<a href="/files/ftp_upload/57388/57388fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
补充文件:补充文件包括. stl 格式的20个文件, 详细说明了超空间引擎组件和配件准备 stereolithographic 打印 (单位毫米) 和1文件中. pdf 格式这是准备加工的车削工具尖端的蓝图。原始的3D 模型文件是用. dwg 格式的软件创建的, 将在请求时可用。<a href="https://cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/57388/supplementary_files_.zip">请单击此处下载此文件.</a>

Watch this Scientific Journal Video about Construction of an Improved Multi-Tetrode Hyperdrive for Large-Scale Neural Recording in Behaving Rats at JoVE.com

Construction of an Improved Multi-Tetrode Hyperdrive for Large-Scale Neural Recording in Behaving Rats

我们提出了一个3维可打印的超空间引擎的建设与十八独立调整 tetrodes。超光速引擎的目的是记录在数周内自由行为的老鼠的大脑活动。

一种改进的多电子四极超光速引擎在行为大鼠大尺度神经记录中的构建

Monitoring the activity patterns of a large population of neurons over many days in awake animals is a valuable technique in the field of systems ...

construction-an-improved-multi-tetrode-hyperdrive-for-large-scale

Universidad San Sebastian

Research

JoVE Journal

Neuroscience

11.2K 次观看.  Baylor College of Medicine. 我们提出了一个3维可打印的超空间引擎的建设与十八独立调整 tetrodes。超光速引擎的目的是记录在数周内自由行为的老鼠的大脑活动。

视频: Construction of an Improved Multi-Tetrode Hyperdrive for Large-Scale Neural Recording in Behaving Rats

Construction of Microdrive Arrays for Chronic Neural Recordings in Awake Behaving Mice

State-of-the-art electrophysiological recordings from the brains of freely behaving animals allow researchers to simultaneously examine local field potentials (LFPs) from populations of neurons and action potentials from individual cells, as the animal engages in experimentally relevant tasks. Chronically implanted microdrives allow for brain recordings to last over periods of several weeks. Miniaturized drives and lightweight components allow for these long-term recordings to occur in small mammals, such as mice. By using tetrodes, which consist of tightly braided bundles of four electrodes in which each wire has a diameter of 12.5 &mu;m, it is possible to isolate physiologically active neurons in superficial brain regions such as the cerebral cortex, dorsal hippocampus, and subiculum, as well as deeper regions such as the striatum and the amygdala. Moreover, this technique insures stable, high-fidelity neural recordings as the animal is challenged with a variety of behavioral tasks. This manuscript describes several techniques that have been optimized to record from the mouse brain. First, we show how to fabricate tetrodes, load them into driveable tubes, and gold-plate their tips in order to reduce their impedance from M&Omega; to K&Omega; range. Second, we show how to construct a custom microdrive assembly for carrying and moving the tetrodes vertically, with the use of inexpensive materials. Third, we show the steps for assembling a commercially available microdrive (Neuralynx VersaDrive) that is designed to carry independently movable tetrodes. Finally, we present representative results of local field potentials and single-unit signals obtained in the dorsal subiculum of mice. These techniques can be easily modified to accommodate different types of electrode arrays and recording schemes in the mouse brain.

The design and assembly of microdrives for in vivo electrophysiological recordings of brain signals from the mouse is described. By attaching microelectrode bundles to sturdy driveable carriers, these techniques allow for long-term and stable neural recordings. The lightweight design allows for unrestricted behavioral performance by the animal following drive implantation.

清醒的行为小鼠慢性神经录音的微型硬盘阵列建设

State-of-the-art electrophysiological recordings from the brains of freely behaving animals allow researchers to simultaneously examine local field ...

科研

行为学

Design and Construction of a Cost Effective Headstage for Simultaneous Neural Stimulation and Recording in the Water Maze

Headstage preamplifiers and source followers are commonly used to study neural activity in behavioral neurophysiology experiments. Available commercial products are often expensive, not easily customized, and not submersible. Here we describe a method to design and build a customized, integrated circuit headstage for simultaneous 4-channel neural recording and 2-channel simulation in awake, behaving animals. The headstage is designed using a free, commercially available CAD-type design package, and can be modified easily to accommodate different scales (e.g. to add channels). A customized printed circuit board is built using surface mount resistors, capacitors and operational amplifiers to construct the unity gain source follower circuit. The headstage is made water-proof with a combination of epoxy, parafilm and a synthetic rubber putty. We have successfully used this device to record local field potentials and stimulate different brain regions simultaneously via independent channels in rats swimming in a water maze. The total cost is &lt; $30/unit and can be manufactured readily.

We present a low-cost method to design and construct a light headstage pre-amplifier system with simultaneous neural recording and stimulation capability. This device can be waterproofed for use in swimming animals.

同时神经刺激，并记录在水迷宫的设计和构建一个具有成本效益探头

Headstage preamplifiers and source followers are commonly used to study neural activity in behavioral neurophysiology experiments. Available commercial ...

神经科学

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer scale. However, current methods mostly rely on tethered cable recordings or only use unidirectional systems, allowing either recording or stimulation of neural activity, but not at the same time or same target. Here, a new wireless, bidirectional device for simultaneous multichannel recording and stimulation of neural activity in freely behaving rats is described. The system operates through a single portable head stage that both transmits recorded activity and can be targeted in real-time for brain stimulation using a telemetry-based multichannel software. The head stage is equipped with a preamplifier and a rechargeable battery, allowing stable long-term recordings or stimulation for up to 1 h. Importantly, the head stage is compact, weighs 12 g (including battery) and thus has minimal impact on the animal&#180;s behavioral repertoire, making the method applicable to a broad set of behavioral tasks. Moreover, the method has the major advantage that the effect of brain stimulation on neural activity and behavior can be measured simultaneously, providing a tool to assess the causal relationships between specific brain activation patterns and behavior. This feature makes the method particularly valuable for the field of deep brain stimulation, allowing precise assessment, monitoring, and adjustment of stimulation parameters during long-term behavioral experiments. The applicability of the system has been validated using the inferior colliculus as a model structure.

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

A wireless, bidirectional system for multi-channel neural recordings and stimulation in freely behaving rats is introduced. The system is light and compact, thus having minimal impact on the animal&#180;s behavioral repertoire. Moreover, this bidirectional system provides a sophisticated tool to assess causal relationships between brain activation patterns and behavior.

用于体内的无线双向接口记录和刺激自由行为大鼠的神经活动

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer ...

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 ...

Micro-drive Array for Chronic in vivo Recording: Drive Fabrication

Chronic recording of large populations of neurons is a valuable technique for studying the function of neuronal circuits in awake behaving rats. Lightweight recording devices carrying a high density array of tetrodes allow for the simultaneous monitoring of the activity of tens to hundreds of individual neurons. Here we describe a protocol for the fabrication of a micro-drive array with twenty one independently movable micro-drives. This device has been used successfully to record from hippocampal and cortical neurons in our lab. We show how to prepare a custom designed, 3-D printed plastic base that will hold the micro-drives. We demonstrate how to construct the individual micro-drives and how to assemble the complete micro-drive array. Further preparation of the drive array for surgical implantation, such as the fabrication of tetrodes, loading of tetrodes into the drive array and gold-plating, is covered in a subsequent video article.

Micro-drive Array for Chronic in vivo Recording: Drive Fabrication

In this protocol we demonstrate how to fabricate a micro-drive array for chronic electrophysiological recordings in rats.

慢性的微型驱动器阵列在体内录音：驱动器的制造

Chronic recording of large populations of neurons is a valuable technique for studying the function of neuronal circuits in awake behaving rats. ...

生物学

Micro-drive Array for Chronic in vivo Recording: Tetrode Assembly

The tetrode, a bundle of four electrodes, has proven to be a valuable tool for the simultaneous recording of multiple neurons in-vivo.  The differential amplitude of action potential signatures over the channels of a tetrode allows for the isolation of single-unit activity from multi-unit signals.  The ability to precisely control the stereotaxic location and depth of the tetrode is critical for studying coordinated neural activity across brain regions.  In combination with a micro-drive array, it is possible to achieve precise placement and stable control of many tetrodes over the course of days to weeks.  In this protocol, we demonstrate how to fabricate and condition tetrodes using basic tools and materials, install the tetrodes into a multi-drive tetrode array for chronic in-vivo recording in the rat, make ground wire connections to the micro-drive array, and attach a protective cone onto the micro-drive array in order to protect the tetrodes from physical contact with the environment.



Micro-drive Array for Chronic in vivo Recording: Tetrode Assembly

In this protocol we demonstrate how to fabricate and condition tetrodes for use with a micro-drive array, which was designed for chronic electrophysiological recordings in rats. In addition, we illustrate the final stages of micro-drive array construction, which includes installing ground wires and a protective cone.

慢性的微型驱动器阵列在体内录音：四极大会

The tetrode, a bundle of four electrodes, has proven to be a valuable tool for the simultaneous recording of multiple neurons in-vivo.  The differential ...

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 ...

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.