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>

<ol>
	<li>O'Keefe, J., Recce, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phase+relationship+between+hippocampal+place+units+and+the+EEG+theta+rhythm.">Phase relationship between hippocampal place units and the EEG theta rhythm.</a> Hippocampus. 3 (3), 317-330 (1993).</li><li>Wilson, M. A., McNaughton, B. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamics+of+the+hippocampal+ensemble+code+for+space.">Dynamics of the hippocampal ensemble code for space.</a> Science. 261 (5124), 1055-1058 (1993).</li><li>Gothard, K. M., Skaggs, W. E., Moore, K. M., McNaughton, B. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Binding+of+hippocampal+CA1+neural+activity+to+multiple+reference+frames+in+a+landmark-based+navigation+task.">Binding of hippocampal CA1 neural activity to multiple reference frames in a landmark-based navigation task.</a> J Neurosci. 16 (2), 823-835 (1996).</li><li>Gray, C. M., Maldonado, P. E., Wilson, M., McNaughton, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tetrodes+markedly+improve+the+reliability+and+yield+of+multiple+single-unit+isolation+from+multi-unit+recordings+in+cat+striate+cortex.">Tetrodes markedly improve the reliability and yield of multiple single-unit isolation from multi-unit recordings in cat striate cortex.</a> J Neurosci Methods. 63 (1-2), 43-54 (1995).</li><li>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=Large-scale+recording+of+neuronal+ensembles.">Large-scale recording of neuronal ensembles.</a> Nat Neurosci. 7 (5), 446-451 (2004).</li><li>Fyhn, M., Hafting, T., Witter, M. P., Moser, E. I., Moser, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Grid+cells+in+mice.">Grid cells in mice.</a> Hippocampus. 18 (12), 1230-1238 (2008).</li><li>Skaggs, W. E., 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=EEG+sharp+waves+and+sparse+ensemble+unit+activity+in+the+macaque+hippocampus.">EEG sharp waves and sparse ensemble unit activity in the macaque hippocampus.</a> J Neurophysiol. 98 (2), 898-910 (2007).</li><li>Guo, P., Pollack, A. J., Varga, A. G., Martin, J. P., Ritzmann, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracellular+wire+tetrode+recording+in+brain+of+freely+walking+insects.">Extracellular wire tetrode recording in brain of freely walking insects.</a> J Vis Exp. (86), (2014).</li><li>Knierim, J. J., McNaughton, B. L., Poe, G. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+spatial+selectivity+of+hippocampal+neurons+during+space+flight.">Three-dimensional spatial selectivity of hippocampal neurons during space flight.</a> Nat Neurosci. 3 (3), 209-210 (2000).</li><li>Leutgeb, S., 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=Independent+codes+for+spatial+and+episodic+memory+in+hippocampal+neuronal+ensembles.">Independent codes for spatial and episodic memory in hippocampal neuronal ensembles.</a> Science. 309 (5734), 619-623 (2005).</li><li>Lansink, C. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+split+microdrive+for+simultaneous+multi-electrode+recordings+from+two+brain+areas+in+awake+small+animals.">A split microdrive for simultaneous multi-electrode recordings from two brain areas in awake small animals.</a> J Neurosci Methods. 162 (1-2), 129-138 (2007).</li><li>Kloosterman, 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=Micro-drive+array+for+chronic+in+vivo+recording:+drive+fabrication.">Micro-drive array for chronic in vivo recording: drive fabrication.</a> J Vis Exp. (26), (2009).</li><li>. Google Patents Available from: <a target="_blank" href="https://www.google.com/patents/US5928143">https://www.google.com/patents/US5928143</a> (1999)</li><li>Redish, A. D., 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=Independence+of+firing+correlates+of+anatomically+proximate+hippocampal+pyramidal+cells.">Independence of firing correlates of anatomically proximate hippocampal pyramidal cells.</a> J Neurosci. 21 (5), RC134 (2001).</li><li>Schmitzer-Torbert, N., Redish, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+activity+in+the+rodent+dorsal+striatum+in+sequential+navigation:+separation+of+spatial+and+reward+responses+on+the+multiple+T+task.">Neuronal activity in the rodent dorsal striatum in sequential navigation: separation of spatial and reward responses on the multiple T task.</a> J Neurophysiol. 91 (5), 2259-2272 (2004).</li><li>Nguyen, D. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Micro-drive+array+for+chronic+in+vivo+recording:+tetrode+assembly.">Micro-drive array for chronic in vivo recording: tetrode assembly.</a> J Vis Exp. (26), (2009).</li><li>Chang, E. H., Frattini, S. A., Robbiati, S., Huerta, P. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Construction+of+microdrive+arrays+for+chronic+neural+recordings+in+awake+behaving+mice.">Construction of microdrive arrays for chronic neural recordings in awake behaving mice.</a> J Vis Exp. (77), e50470 (2013).</li><li>Vandecasteele, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Large-scale+recording+of+neurons+by+movable+silicon+probes+in+behaving+rodents.">Large-scale recording of neurons by movable silicon probes in behaving rodents.</a> J Vis Exp. (61), e3568 (2012).</li><li>Siegle, J. H., 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=Chronically+implanted+hyperdrive+for+cortical+recording+and+optogenetic+control+in+behaving+mice.">Chronically implanted hyperdrive for cortical recording and optogenetic control in behaving mice.</a> Conf Proc IEEE Eng Med Biol Soc. 2011, 7529-7532 (2011).</li><li>Brunetti, P. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+and+fabrication+of+ultralight+weight,+adjustable+multi-electrode+probes+for+electrophysiological+recordings+in+mice.">Design and fabrication of ultralight weight, adjustable multi-electrode probes for electrophysiological recordings in mice.</a> J Vis Exp. (91), e51675 (2014).</li><li>. Google Patents Available from: <a target="_blank" href="https://www.google.com/patents/US4575330">https://www.google.com/patents/US4575330</a> (1986)</li><li>Ludvig, N., Potter, P. E., Fox, S. 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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...

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&#34; in diameter</td>
		</tr>
		<tr>
			<td>Screw</td>
			<td>McMaster</td>
			<td>91771A060</td>
			<td>Stainless steel, flat head, 0-80&#160;thread,&#160;5/8&#34;&#160;in length</td>
		</tr>
		<tr>
			<td>Screw</td>
			<td>McMaster</td>
			<td>91772A051</td>
			<td>Stainless steel, pan head, 0-80&#160;thread,&#160;5/32&#34;&#160;in length</td>
		</tr>
		<tr>
			<td>Screw</td>
			<td>McMaster</td>
			<td>92196A056</td>
			<td>Stainless steel, socket head, 0-80&#160;thread,&#160;5/16&#34;&#160;in length</td>
		</tr>
		<tr>
			<td>Screw</td>
			<td>McMaster</td>
			<td>92196A055</td>
			<td>Stainless steel, socket head, 0-80&#160;thread,&#160;1/4&#34;&#160;in length</td>
		</tr>
		<tr>
			<td>Screw</td>
			<td>McMaster</td>
			<td>95868A131</td>
			<td>Nylon,&#160; socket head, 2-56&#160;thread,&#160;3/16&#34;&#160;in length, black</td>
		</tr>
		<tr>
			<td>Screw nut</td>
			<td>McMaster</td>
			<td>90730A001</td>
			<td>Stainless steel, narrow hex,&#160; 0-80&#160;thread</td>
		</tr>
		<tr>
			<td>Shoulder screw</td>
			<td>McMaster</td>
			<td>90298A213</td>
			<td>Stainless steel, 8-32 thread, 3/16&#34; in diameter, 1/4&#34; in length</td>
		</tr>
		<tr>
			<td>Cup screw</td>
			<td>McMaster</td>
			<td>92313A105</td>
			<td>Stainless steel, 4-40 thread, 3/16&#34; in length</td>
		</tr>
		<tr>
			<td>Thumb screw</td>
			<td>McMaster</td>
			<td>94323A592</td>
			<td>Nylon, 8-32&#160;thread,&#160;3/8&#34;&#160;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&#34; OD, 0.013&#34; ID, 0.006&#34; 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&#34; OD, 0.007/0.008&#34; ID, full hard</td>
		</tr>
		<tr>
			<td>Heat-shrink tubing</td>
			<td>McMaster</td>
			<td>7856K72</td>
			<td>0.09&#34; ID before shrinking, blue</td>
		</tr>
		<tr>
			<td>Silicone tubing</td>
			<td>A-M Systems</td>
			<td>807300</td>
			<td>0.040&#34; ID, 0.085&#34; OD</td>
		</tr>
		<tr>
			<td>Polyimide tubing</td>
			<td>A-M Systems</td>
			<td>823400</td>
			<td>0.0045&#34; ID, 0.0005&#34; wall</td>
		</tr>
		<tr>
			<td>Ground wire</td>
			<td>A-M Systems</td>
			<td>791500</td>
			<td>0.005&#34; bare, 0.008&#34; coated, half hard</td>
		</tr>
		<tr>
			<td>Tetrode wire</td>
			<td>California Fine Wire</td>
			<td>Custom-made</td>
			<td>0.0007&#34; 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&#34;-24&#160;thread size, bottoming</td>
		</tr>
		<tr>
			<td>Tap</td>
			<td>McMaster</td>
			<td>2522A713</td>
			<td>2-56&#160;thread size</td>
		</tr>
		<tr>
			<td>Tap</td>
			<td>McMaster</td>
			<td>2522A715</td>
			<td>4-40&#160;thread size</td>
		</tr>
		<tr>
			<td>Tap</td>
			<td>McMaster</td>
			<td>2522A718</td>
			<td>8-32&#160;thread size</td>
		</tr>
		<tr>
			<td>Die</td>
			<td>McMaster</td>
			<td>2576A457</td>
			<td>3/8&#34;-24&#160;thread size, 1&#34; OD</td>
		</tr>
		<tr>
			<td>Drill bit</td>
			<td>McMaster</td>
			<td>30585A82</td>
			<td>Wire gauge&#160;65,&#160;0.035&#34; in diameter</td>
		</tr>
		<tr>
			<td>Drill bit</td>
			<td>McMaster</td>
			<td>30585A83</td>
			<td>Wire gauge&#160;66,&#160;0.033&#34; in diameter</td>
		</tr>
		<tr>
			<td>Drill bit</td>
			<td>McMaster</td>
			<td>30585A87</td>
			<td>Wire gauge&#160;70,&#160;0.028&#34; in diameter</td>
		</tr>
		<tr>
			<td>Drill bit</td>
			<td>McMaster</td>
			<td>30585A88</td>
			<td>Wire gauge&#160;71,&#160;0.026&#34; in diameter</td>
		</tr>
		<tr>
			<td>Drill bit</td>
			<td>McMaster</td>
			<td>30585A91</td>
			<td>Wire gauge&#160;73,&#160;0.024&#34; in diameter</td>
		</tr>
		<tr>
			<td>Drill bit</td>
			<td>McMaster</td>
			<td>8870A23</td>
			<td>3/16&#34;&#160;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&#34; in diameter, full hard</td>
		</tr>
		<tr>
			<td>Steel wire</td>
			<td>Precision Brand</td>
			<td>21007</td>
			<td>0.007&#34; in diameter, full hard</td>
		</tr>
		<tr>
			<td>Steel wire</td>
			<td>A-M Systems</td>
			<td>792700</td>
			<td>0.003&#34; 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&#160;</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?...

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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

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

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 ...

科研

神经科学

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 ...

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 ...

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 ...

Preparation of an Awake Mouse for Recording Neural Responses and Injecting Tracers

It is well known that anesthesia alters neural response properties in various regions of the brain.13. In the auditory system, fundamental response properties of brainstem neurons including threshold, frequency specificity, and inhibitory sidebands are altered in significant ways under anesthesia1-2. These observations prompted physiologists to seek ways to record from single neurons without the contaminating effects of anesthesia. One result was a decerebrate preparation, where the brainstem was completely transected at the level of the midbrain4. The drawbacks of this preparation are a formidable surgery, the elimination of descending projections from the forebrain, and an inability to use sensory stimulation to examine structures above the midbrain. A different strategy has been to implant electrode arrays chronically to record from single neurons and multiunit clusters while the animal is awake and/or behaving5,6. These techniques however are not compatible with injecting tracer dyes after first electrophysiologically characterizing a brain structure. To avoid altering neural response properties with anesthetics while recording electrophysiological response properties from single neurons, we have adapted a head restraint technique long used in bats7-9 to mouse10-12. Using this method, we are able to conduct electrophysiological recordings over several days in the unanesthetized mouse. At the end of the recording sessions, we can then inject a dye to reconstruct electrode positions and recording sites or inject a tracer so that pathways to and from the recording loci can be determined. This method allows for well isolated single neuron recordings over multiple days without the use anesthetics.

Electrophysiological characterization of neuronal responses is important for understanding brain function and for guiding the placement of dyes for pathway tracing. However, many studies are performed in anesthetized animals. To understand brain function without anesthetics, we developed a method to record neuronal response properties and inject dyes in awake mouse.

记录神经反应和注射示踪剂一个清醒小鼠的制备

It is well known that anesthesia alters neural response properties in various regions of the brain.13.  In the auditory system, fundamental response ...

Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

Detection and interpretation of olfactory cues are critical for the survival of many organisms. Remarkably, species across phyla have strikingly similar olfactory systems suggesting that the biological approach to chemical sensing has been optimized over evolutionary time1. In the insect olfactory system, odorants are transduced by olfactory receptor neurons (ORN) in the antenna, which convert chemical stimuli into trains of action potentials. Sensory input from the ORNs is then relayed to the antennal lobe (AL; a structure analogous to the vertebrate olfactory bulb). In the AL, neural representations for odors take the form of spatiotemporal firing patterns distributed across ensembles of principal neurons (PNs; also referred to as projection neurons)2,3. The AL output is subsequently processed by Kenyon cells (KCs) in the downstream mushroom body (MB), a structure associated with olfactory memory and learning4,5. Here, we present electrophysiological recording techniques to monitor odor-evoked neural responses in these olfactory circuits.
First, we present a single sensillum recording method to study odor-evoked responses at the level of populations of ORNs6,7. We discuss the use of saline filled sharpened glass pipettes as electrodes to extracellularly monitor ORN responses. Next, we present a method to extracellularly monitor PN responses using a commercial 16-channel electrode3. A similar approach using a custom-made 8-channel twisted wire tetrode is demonstrated for Kenyon cell recordings8. We provide details of our experimental setup and present representative recording traces for each of these techniques.

Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

We demonstrate variations of the extracellular multi-unit recording technique to characterize odor-evoked responses in the first three stages of the invertebrate olfactory pathway. These techniques can easily be adapted to examine ensemble activity in other neural systems as well.

多单位记录方法来描述蝗虫的神经活动（<em&gt; Schistocerca美洲</em&gt;）嗅觉电路

Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

Neural Circuit Recording from an Intact Cockroach Nervous System

The cockroach ventral nerve cord preparation is a tractable system for neuroethology experiments, neural network modeling, and testing the physiological effects of insecticides. This article describes the scope of cockroach sensory modalities that can be used to assay how an insect nervous system responds to environmental perturbations. Emphasis here is on the escape behavior mediated by cerci to giant fiber transmission in Periplaneta americana. This in situ preparation requires only moderate dissecting skill and electrophysiological expertise to generate reproducible recordings of neuronal activity. Peptides or other chemical reagents can then be applied directly to the nervous system in solution with the physiological saline. Insecticides could also be administered prior to dissection and the escape circuit can serve as a proxy for the excitable state of the central nervous system. In this context the assays described herein would also be useful to researchers interested in limb regeneration and the evolution of nervous system development for which P. americana is an established model organism.

This article describes the cockroach ventral nerve cord dissection and extracellular recordings from the cercal nerve and connectives. Evoked responses are generated by electrical stimulation of the cercal nerve or direct mechanical stimulation of the cerci.

神经电路记录从一个完整的蟑螂神经系统

The cockroach ventral nerve cord preparation is a tractable system for neuroethology experiments, neural network modeling, and testing the physiological ...

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method described here enables the investigator to measure long-lasting (from minutes to hours) high-quality intracellular responses from single Drosophila R1-R6 photoreceptors and Large Monopolar Cells (LMCs) to light stimuli. Because the recording system has low noise, it can be used to study variability among individual cells in the fly eye, and how their outputs reflect the physical properties of the visual environment. We outline all key steps in performing this technique. The basic steps in constructing an appropriate electrophysiology set-up for recording, such as design and selection of the experimental equipment are described. We also explain how to prepare for recording by making appropriate (sharp) recording and (blunt) reference electrodes. Details are given on how to fix an intact fly in a bespoke fly-holder, prepare a small window in its eye and insert a recording electrode through this hole with minimal damage. We explain how to localize the center of a cell's receptive field, dark- or light-adapt the studied cell, and to record its voltage responses to dynamic light stimuli. Finally, we describe the criteria for stable normal recordings, show characteristic high-quality voltage responses of individual cells to different light stimuli, and briefly define how to quantify their signaling performance. Many aspects of the method are technically challenging and require practice and patience to master. But once learned and optimized for the investigator's experimental objectives, it grants outstanding in vivo neurophysiological data.

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Sharp microelectrodes enable accurate electrophysiological characterization of photoreceptor and visual interneuron output in living Drosophila. Here we show how to use this method to record high-quality voltage responses of individual cells to controlled light stimulation. This method is ideal for studying neural information processing in insect compound eyes.

为记录细胞内电压响应电法<em&gt;果蝇</em&gt;光感受器和中间对光刺激<em&gt;在体内</em

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method ...

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 ...

Large-scale Three-dimensional Imaging of Cellular Organization in the Mouse Neocortex

The mammalian neocortex is composed of many types of excitatory and inhibitory neurons, each with specific electrophysiological and biochemical properties, synaptic connections, and in vivo functions, but their basic functional and anatomical organization from cellular to network scale is poorly understood. Here we describe a method for the three-dimensional imaging of fluorescently-labeled neurons across large areas of the brain for the investigation of the cortical cellular organization. Specific types of neurons are labeled by the injection of fluorescent retrograde neuronal tracers or expression of fluorescent proteins in transgenic mice. Block brain samples, e.g., a hemisphere, are prepared after fixation, made transparent with tissue clearing methods, and subjected to fluorescent immunolabeling of the specific cell types. Large areas are scanned using confocal or two-photon microscopes equipped with large working distance objectives and motorized stages. This method can resolve the periodic organization of the cell type-specific microcolumn functional modules in the mouse neocortex. The procedure can be useful for the study of three-dimensional cellular architecture in the diverse brain areas and other complex tissues.

Here we describe a procedure for tissue clearing, fluorescent labeling, and large-scale imaging of mouse brain tissue which, thereby, enables visualization of the three-dimensional organization of cell types in the neocortex.