Name: wireless electrophysiological recording of neurons by movable tetrodes in freely swimming fish
Uploaded: 2019-11-26T15:00:00
Description: Watch this Scientific Journal Video about Wireless Electrophysiological Recording of Neurons by Movable Tetrodes in Freely Swimming Fish at JoVE.com

我们感谢纳楚姆·乌拉诺夫斯基和乌拉诺夫斯基实验室的成员提供的所有帮助。此外，我们感谢塔尔·诺普兰斯基-祖尔提供有益的技术援助。我们感谢来自以色列科学基金会-第一计划（第281/15号赠款）和赫尔姆斯利慈善信托基金通过内盖夫本-古里安大学的农业、生物和认知机器人倡议提供的财政支持。

所有手术程序都必须得到当地动物福利伦理委员会（如IACUC）的批准。
1. 微型驱动外壳的建造
<ol><li>要构建外壳，请使用锯片将 1 mm 宽的黄铜板切割成 19 mm x 29 mm x 1 mm 板。在垂直于边缘的两条长边上切割两个 5.5 mm 的狭缝，使每个狭缝距离窄边 6.5 mm（图2A）。</li>
	<li>使用钳子，将长边上长边之间的区域向内折叠，然后将底部向内折叠，将...

<ol>
	<li>Jacobson, M., Gaze, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Types+of+visual+response+from+single+units+in+the+optic+tectum+and+optic+nerve+of+the+goldfish.">Types of visual response from single units in the optic tectum and optic nerve of the goldfish.</a> Quarterly Journal of Experimental Physiology and Cognate Medical Sciences. 49 (2), 199-209 (1964).</li><li>Ben-Tov, M., Donchin, O., Ben-Shahar, O., Segev, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pop-out+in+visual+search+of+moving+targets+in+the+archer+fish.">Pop-out in visual search of moving targets in the archer fish.</a> Nature Communications. 6, 6476 (2015).</li><li>Zottoli, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Correlation+of+the+startle+reflex+and+Mauthner+cell+auditory+responses+in+unrestrained+goldfish.">Correlation of the startle reflex and Mauthner cell auditory responses in unrestrained goldfish.</a> Journal of Experimental Biology. 66 (1), 243-254 (1977).</li><li>Canfield, J. G., Rose, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activation+of+Mauthner+neurons+during+prey+capture.">Activation of Mauthner neurons during prey capture.</a> Journal of Comparative Physiology A. 172 (5), 611-618 (1993).</li><li>Canfield, J. G., Mizumori, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+for+chronic+neural+recording+in+the+telencephalon+of+freely+behaving+fish.">Methods for chronic neural recording in the telencephalon of freely behaving fish.</a> Journal of Neuroscience Methods. 133 (1-2), 127-134 (2004).</li><li>Orger, M. B., de Polavieja, G. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zebrafish+behavior:+opportunities+and+challenges.">Zebrafish behavior: opportunities and challenges.</a> Annual Review of Neuroscience. 40, 125-147 (2017).</li><li>Vanwalleghem, G. C., Ahrens, M. B., Scott, E. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integrative+whole-brain+neuroscience+in+larval+zebrafish.">Integrative whole-brain neuroscience in larval zebrafish.</a> Current Opinion in Neurobiology. 50, 136-145 (2018).</li><li>Vinepinsky, E., Donchin, O., Segev, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wireless+electrophysiology+of+the+brain+of+freely+swimming+goldfish.">Wireless electrophysiology of the brain of freely swimming goldfish.</a> Journal of Neuroscience Methods. 278, 76-86 (2017).</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> JoVE (Journal of Visualized Experiments). (61), e3568 (2012).</li><li>Ferguson, J. E., Boldt, C., 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=Creating+low-impedance+tetrodes+by+electroplating+with+additives.">Creating low-impedance tetrodes by electroplating with additives.</a> Sensors and Actuators A: Physical. 156 (2), 388-393 (2009).</li><li>Arcot Desai, S., Rolston, J. D., Guo, L., Potter, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improving+impedance+of+implantable+microwire+multi-electrode+arrays+by+ultrasonic+electroplating+of+durable+platinum+black.">Improving impedance of implantable microwire multi-electrode arrays by ultrasonic electroplating of durable platinum black.</a> Frontiers in Neuroengineering. 3, 5 (2010).</li><li>Lewicki, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+methods+for+spike+sorting:+the+detection+and+classification+of+neural+action+potentials.">A review of methods for spike sorting: the detection and classification of neural action potentials.</a> Network: Computation in Neural Systems. 9 (4), R53-R78 (1998).</li><li>Teixeira, F. B., Freitas, P., Pessoa, L. M., Campos, R. L., Ricardo, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+IEEE+802.11+underwater+networks+operating+at+700+MHz,+2.4+GHz+and+5+GHz.">Evaluation of IEEE 802.11 underwater networks operating at 700 MHz, 2.4 GHz and 5 GHz.</a> Proceedings of the 10th International Conference on Underwater Networks &amp; Systems. , (2015).</li><li>Sendra, S., Lloret, J., Rodrigues, J. J., Aguiar, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Underwater+wireless+communications+in+freshwater+at+2.4+GHz.">Underwater wireless communications in freshwater at 2.4 GHz.</a> IEEE Communications Letters. 17 (9), 1794-1797 (2013).</li><li>Lloret, J., Sendra, S., Ardid, M., Rodrigues, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Underwater+wireless+sensor+communications+in+the+2.4+GHz+ISM+frequency+band.">Underwater wireless sensor communications in the 2.4 GHz ISM frequency band.</a> Sensors. 12 (4), 4237-4264 (2012).</li><li>Hoogerwerf, A. C., Wise, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+three-dimensional+microelectrode+array+for+chronic+neural+recording.">A three-dimensional microelectrode array for chronic neural recording.</a> IEEE Transactions on Biomedical Engineering. 41 (12), 1136-1146 (1994).</li><li>Harris, K. D., Quiroga, R. Q., Freeman, J., Smith, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improving+data+quality+in+neuronal+population+recordings.">Improving data quality in neuronal population recordings.</a> Nature Neuroscience. 19 (9), 1165 (2016).</li></ol>

该协议详细说明了在自由游泳的金鱼的端脑中植入ttrode阵列所涉及的步骤。该技术实现了一个神经记录器，放大和记录从多达16个通道获得的信号，以及一个可以调整大脑中的ttrode位置的微驱。微驱可以调整大脑中的位置以优化录制。
该协议可以很容易地修改，以便从其他大脑区域进行记录（参见Vinepinsky等人8，以便使用类似的技术从光学技术中记录）或任?...

鱼类是最大、最多样化的脊椎动物群，与其他脊椎动物一样，它们表现出复杂的认知能力，如导航、社交、睡眠、狩猎等。然而，控制鱼类行为的神经机制在很大程度上仍不为人所知。
在过去的几十年里，固定鱼的细胞外记录主要用于研究行为1、2的神经基础的不同方面。虽然这种技术适用于某些感官系统，但研究行为的神经基础的全部谱是很难的，如果不是不可能在固定动物。第一个进展涉及记录从毛斯纳细胞的系绳游泳鱼3，4。然而，毛斯纳细胞不成比例地大，记录的动作潜力振幅，可以高达几mV，便于记录。后来，Canfield等人描述了一个概念证明，当使用系绳动物记录从鱼的telencephon5。记录鱼类神经活动的另一项最新技术是钙成像（见奥尔格和德波拉维娅6和Vanwalleghem等人的评论）。这项技术是开发用于斑马鱼幼虫，因为皮肤和头骨是透明的，在幼虫阶段。然而，这种技术不能用于研究后期开发阶段的复杂行为。
在这里，我们提出了一种新技术，用于记录自由游泳鱼大脑中的细胞外神经活动。这是Vinepinsky等人8中描述的协议的修改版本。主要的创新是增加了一个微驱，使得手术后能够控制电极的位置。该技术设计用于使用一组通过微驱动器连接到神经数据记录器的四叶鱼记录金鱼的端管。整个设置是无线的，并锚定在鱼的头骨。系统的特定重量通过添加一个小浮子，使鱼可以自由游泳，从而与水特定重量相等。
该技术基于使用神经数据记录器，该记录器放大、数字化并将信号存储在板载存储设备中。记录器遥测系统用于启动和停止录像，以及与摄像机同步。在此协议中，使用 16 通道神经记录器，与微型驱动器一起嵌入防水盒中。
微驱组件由两个主要部件制成：微驱本身和微驱外壳（图1A，B）。外壳装有微驱动器和三头龙，还充当头骨和记录器箱之间的锚点（图1C）。PVC记录器盒采用机器工艺制造，使用 O 形环进行密封（图1E-G，另见补充图 1、补充图 2和辅助图 3，用于三维 [3D] 图）。在一端，一块聚苯乙烯泡沫被连接到记录器盒上，以补偿植入物的重量，并为鱼提供浮力中性植入物。协议中描述的微驱的构造遵循 Vandecasteele 等人9提出的程序，对微驱连接到外壳进行了修改（图 1A）。介绍了所有主要步骤。
议定书中描述的鱼头骨制备程序与Vinepinsky等人8中所述的程序类似，并在协议中作了简要说明。手术后一天，鱼通常从麻醉效果中完全恢复，并准备进行行为实验。请注意，可以通过转动微驱螺钉来调整 ttrode 位置。螺钉每次完全旋转的间距为 300 μm，建议在达到目标大脑位置之前提高 75 μm。应参考适当的大脑图集，以针对感兴趣的特定大脑区域。建议每次对鱼进行麻醉以更换电池或存储卡时测试电极阻抗。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.7 mm round drill bits</td>
			<td></td>
			<td></td>
			<td>Compatible with the drill.</td>
		</tr>
		<tr>
			<td>15-blade Scalpel</td>
			<td>Sigma-Aldrich</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>16 channel PCB board</td>
			<td>Neurlynx</td>
			<td>EIB-16</td>
			<td></td>
		</tr>
		<tr>
			<td>1X3M phillips flat head screws</td>
			<td></td>
			<td></td>
			<td>Stainless steel. Any type.</td>
		</tr>
		<tr>
			<td>1X3M phillips round head screws</td>
			<td></td>
			<td></td>
			<td>Stainless steel. Any type.</td>
		</tr>
		<tr>
			<td>27 cm X 19 cm X 1 mm brass plate</td>
			<td></td>
			<td></td>
			<td>See Figure 2</td>
		</tr>
		<tr>
			<td>2X6M phillips flat head screws</td>
			<td></td>
			<td></td>
			<td>Stainless steel. Any type.</td>
		</tr>
		<tr>
			<td>3140 RTV coating</td>
			<td>Dow Crowning</td>
			<td>2767996</td>
			<td></td>
		</tr>
		<tr>
			<td>75 &#181;m Silver wire</td>
			<td>A-M Systems</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Brass machine screws #00-90</td>
			<td></td>
			<td>947-1006</td>
			<td></td>
		</tr>
		<tr>
			<td>Brass plates 7.5mm X 2.5mm X 0.6mm</td>
			<td></td>
			<td></td>
			<td>A 3D drawing is provided. See supplementary 1</td>
		</tr>
		<tr>
			<td>Coated Tungsten wire 25&#181;m</td>
			<td>California Fine Wire Company</td>
			<td>5000160</td>
			<td>Depending on the appication the tetrodes can be fabricated from any type of wire. Popular wires are nicrome wires that can be found with lower diameters (eg. A-M systems, 762000)</td>
		</tr>
		<tr>
			<td>Coated Tungsten wire 50&#181;m</td>
			<td>A-M Systems</td>
			<td>795500</td>
			<td>Can be replaced with any other wire with low impedance</td>
		</tr>
		<tr>
			<td>Cyanoacrilic glue</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dental Burnisher</td>
			<td>ComDent UK</td>
			<td></td>
			<td>Any small sterille stainless-still tool will do.</td>
		</tr>
		<tr>
			<td>Dental cement - GCFujiPLUS</td>
			<td>GC</td>
			<td>431011</td>
			<td>Other dental cements would probably will work as well although we have never tried any other.</td>
		</tr>
		<tr>
			<td>Dental drill or nail polish drill</td>
			<td></td>
			<td></td>
			<td>Dental drills are expensive, a nail polish drill can be a cheap replacement.</td>
		</tr>
		<tr>
			<td>Drill bit #65</td>
			<td></td>
			<td>947-65</td>
			<td></td>
		</tr>
		<tr>
			<td>Fast curing epoxy</td>
			<td></td>
			<td></td>
			<td>Any 5 minutes curing epoxy can be used here.</td>
		</tr>
		<tr>
			<td>Logger box with O-ring sealing</td>
			<td></td>
			<td></td>
			<td>A 3D drawing is provided. See supplementary 1-3. The box should be machine fabricated (do not use 3D printers). Use transperant material, to be able to see the indicator LEDs on the logger.</td>
		</tr>
		<tr>
			<td>Motorized turning device</td>
			<td></td>
			<td></td>
			<td>Custom made as described in &#34;open ephys&#34; website. Can also be purchusaed from neurolynx (&#34;Tetrode Spinner 2.0&#34;) or bulit by other means.</td>
		</tr>
		<tr>
			<td>Mouselog-16 Neural logger</td>
			<td>Deuteron Technologies Ltd</td>
			<td></td>
			<td>There are several neural loggers available on the market, including: SpikeGadget (UH32 32channels) and Neurologger 2/2A/2B of Alexei Vyssotski. It should be noted that weight is not a major contraint since it can be counterbalanced with floating Styrofoam</td>
		</tr>
		<tr>
			<td>MS-222</td>
			<td>Sigma Aldrich</td>
			<td>E10521</td>
			<td>Ethtl 3-aminobenzoate methanesulfonate 98%</td>
		</tr>
		<tr>
			<td>Nano-Z plating</td>
			<td>White Matter LLC</td>
			<td></td>
			<td>The nano-Z can be bought from several supllieres. Any impedance meter can be used, e.g. IMP-1 / 6662 / 2788, BAK Electronics.</td>
		</tr>
		<tr>
			<td>PCB pins</td>
			<td>Neurlynx</td>
			<td>Neuralynx EIB Pins</td>
			<td></td>
		</tr>
		<tr>
			<td>Polymide tubing 250&#181;m</td>
			<td>A-M Systems</td>
			<td>822000</td>
			<td></td>
		</tr>
		<tr>
			<td>Rechargable battery</td>
			<td></td>
			<td></td>
			<td>3.7 Lipo battery, 370 mAh. Holds about 6 hours of recording. Smaller or larger battries can be used to reduce the weight or extend recording time.</td>
		</tr>
		<tr>
			<td>Silicone tubing 0.64 mm</td>
			<td>A-M Systems</td>
			<td>806100</td>
			<td></td>
		</tr>
		<tr>
			<td>Stainless steel 1.5 mm</td>
			<td>A-M Systems</td>
			<td>846000</td>
			<td></td>
		</tr>
		<tr>
			<td>Sudium Bicarbonate</td>
			<td>Sigma Aldrich</td>
			<td>S9625</td>
			<td></td>
		</tr>
		<tr>
			<td>Tap #00-90</td>
			<td></td>
			<td>947-1301</td>
			<td></td>
		</tr>
		<tr>
			<td>Vaseline</td>
			<td></td>
			<td></td>
			<td>Any type of soft petroleum skin protectant can be used here.</td>
		</tr>
	</tbody>
</table>

wireless electrophysiological recording of neurons by movable tetrodes in freely swimming fish

控制鱼类行为的神经机制仍然鲜为人知，尽管鱼类占所有脊椎动物的大多数。记录自由移动鱼类的大脑活动的能力将大大促进对鱼类行为的神经基础的研究。此外，精确控制大脑中的记录位置对于研究鱼脑中跨区域的协调神经活动至关重要。在这里，我们介绍一种技术，从自由游泳的鱼的大脑无线记录，同时控制记录位置的深度。该系统基于与新型水兼容植入物相关的神经记录器，该植入物可以通过微驱控制三轮驱动来调整记录位置。该系统的功能通过金鱼的端管记录来说明。

在记录过程中，金鱼在方形水箱中自由游动，而其端脑的神经活动被记录下来。这些实验的目的是研究单个细胞的神经活动如何决定鱼的行为。为此，需要在记录的数据中确定尖峰活动。大脑活动在被记录时，被数字化为31，250赫兹，高通过滤在300赫兹的数据记录器。然后，离线，对信号应用了带通滤波器（300~5，000 Hz），并将预排序的原始数据分离到每个ttrode的通道和参...

Watch this Scientific Journal Video about Wireless Electrophysiological Recording of Neurons by Movable Tetrodes in Freely Swimming Fish at JoVE.com

Wireless Electrophysiological Recording of Neurons by Movable Tetrodes in Freely Swimming Fish

介绍了一种记录自由游泳金鱼大脑细胞外神经信号的新型无线技术。记录装置由两个三元仪组成，一个微型驱动器，一个神经数据记录器和一个防水外壳。除数据记录器及其连接器外，所有部件都是定制的。

自由游泳鱼中可移动的特龙对神经元的无线电生理记录

The neural mechanisms governing fish behavior remain mostly unknown, although fish constitute the majority of all vertebrates. The ability to record brain ...

我们开发了一种从自由游泳的鱼的大脑记录细胞外神经信号的新方法。这为研究一天末鱼类行为的神经元机制打开了大门。这种方法是有效的，防水，因此允许记录，而鱼在水箱中自由游泳。此外，记录位置可由内置微驱动器控制。我们的实验室技术员塔尔·祖尔-诺沃普兰斯基将帮助我演示这个程序。要建造外壳，请使用尺寸为 19 毫米、29 毫米和 1 毫米的黄铜板。长边有两个 5.5 毫米的狭缝，垂直于边缘。狭缝应远离狭窄的一侧 6.5 毫米。使用钳子将长边上的狭缝之间的区域向内折叠。然后向内折叠底部和上侧向外，以获得外壳。接下来，使用三毫米迪尔位在微驱动壳体上为螺钉打孔，并焊接壳体两侧。使用精细的圆形文件在外壳底部创建半径为 1.5 毫米的小半圆形狭缝。使用一毫米钻头在壳体背面为四孔打一个洞。首先，使用刀具从单排公针头条上分离出三根针片。使用钳子拉出中间销。接下来，使用刀具将剩余引脚切割至 10 毫米的长度。使用数字 65 钻头在中间销孔中钻孔，并使用双零 99 水龙头钻孔。组装微型驱动器和黄铜板。通过第一个黄铜板插入螺钉，通过销头螺纹，然后通过第二个黄铜板。最后，在螺钉上放置螺母，轻轻拧紧组装的微型驱动器。将销与黄铜板一起焊接，将螺母与螺钉尖端一起焊接。在此之后，将微型驱动器焊接到微型驱动器黄铜板两侧的四个点上的微驱动器壳体中。使用环氧胶水六毫米长的不锈钢管到微型驱动器外壳底部的小半圆形狭缝。将三毫米长的不锈钢管粘在销头上，使固定壳体上与六毫米长的管子排成一排。将准备好的硅胶和聚酰亚胺管插入两个不锈钢管，并使用氰酸酯胶水将它们粘在连接到针头的不锈钢偷管上。然后，将微型驱动器一直拧上，从两根钢管的顶部和底部切断多余的管子。接下来，切割两根直径为 75 微米、长 12 厘米的裸银线，并将它们焊接到电极接口板中的接地连接和所有未使用的通道上。将其中一根准备好的钨丝推入 16 通道电极接口板的孔中。将针放在孔中，用一对钳子按压。测量导线的引脚和未编码端之间的电阻，以检查连接性。对所有准备好的导线（包括一根 50 微米导线）重复此过程，用于参考电极。然后将电极线分成两组，每组四根导线，然后用每根导线的端端的胶带将它们缠绕在一起，使参考导线单独使用。将电极接口板放在电动车削装置上方，然后将胶带端从一组四根导线放在设备上。顺时针应用 130 轮，然后顺时针旋转 20 轮。然后，涂抹氰丙烯酸酯胶水以覆盖四边形。胶水固化后，将四甲靠近胶带切开。将旋转、粘合和切割过程重复到其他四边体。首先，用两毫米螺钉将微型驱动器壳体连接到记录箱。通过微驱动壳体背面的孔螺纹四头和参考电极。通过两个硅胶管螺纹四管，通过聚酰亚胺管螺纹 50 微米钨丝。拧微潜水一路下来，并应用氰丙烯酸酯胶水到管的顶部，以粘合的四边形和电线到他们的管子，并确保运动与微驱动器一致。将微型驱动器一直拧到顶部。将软石油涂抹在微型驱动器壳体内的外露四头和电线上，以防止运动。将预切培养皿底部窗口连接到带环氧树脂的微型驱动器壳体前部，同时确保将接地线放在车窗外。接下来，在记录箱盖和微驱动器外壳之间的外露四头板和电线上涂上室温硫化涂层。使用锋利的剪刀将四端和参考线切割到所需长度。在此之后，将标记的、散发的聚苯乙烯泡沫连接到包装盒中。调整其大小，使浮力在浸入水浴中时保持平衡。麻醉鱼后，把鱼从水里拿出来，放在鱼架上。使用无菌铲子在指定手术地点上方的皮肤上涂抹 5% 的利多卡因膏 10 分钟。然后取出利多卡因。使用无菌编号 15 刀片手术刀去除为植入物指定的区域中头骨上方的皮肤。然后使用带 0.7 毫米钻头的牙科钻头在头骨上钻四个孔。将氰丙烯酸酯胶涂抹到其中一个孔上，并立即插入一毫米螺钉。对剩余的钻孔重复此过程。在此之后，使用牙科烧制剂在螺钉和暴露的头骨的外围涂抹牙科水泥。使用牙科钻头打一个直径超过大脑区域五毫米的孔。使用细钳子和软纸巾去除头骨和大脑之间的脂肪组织，并暴露大脑区域目标小心不要损坏头骨下面的大血管。首先，降低植入物，使电极插入大脑，而微驱动器壳体的底部靠近头骨。在壳体和最近的头骨螺钉之间涂抹少量牙科水泥，开始将植入物连接到头骨上。牙科水泥的第一部分固化后，添加牙科水泥，并关闭头骨上方的孔和整个暴露的头骨。用淡水冲洗鱼的刺来唤醒鱼的刺后，将鱼从鱼架上取出，放回家中的水箱中。确保鱼能够自由游泳与植入物。这些实验的目标是研究单细胞的神经活动如何编码鱼的行为。为此，将四边甲阵列植入自由游泳金鱼的电头，从而可以记录神经活动。记录时的大脑活动被数字化为31，250赫兹和高通过滤在300赫兹的数据记录器。然后离线将禁止通滤波器应用于信号，将预排序的原始数据分离到每个四通的通道和参考通道中。下一个常见的尖峰排序算法用于描述单个单元活动。首先，每个通道按最小峰值振幅阈值手动筛选。在多个四分中或参考通道中出现的尖峰也会被过滤。然后，通过形状、长度、尖峰间间隔和原理分量分析手动聚类和筛选检测到的峰值。在手术前检查植入物的完整性非常重要。可以使用标准阻抗计检查四管一，并且应检查箱体在水下有无泄漏。这种方法可以修改，以工作与许多水生动物，只要他们足够大，以容纳系统。这使得研究其行为背后的神经机制。

Research

JoVE Journal

Neuroscience

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

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 ...

科研

神经科学

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

逆行荧光染料标记允许有针对性的外单细胞记录技术鉴定神经元<em&gt;在体内</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

Forebrain Electrophysiological Recording in Larval Zebrafish

斑马鱼幼虫的前脑的电生理记录

Epilepsy affects nearly 3 million people in the United States and up to 50 million people worldwide. Defined as the occurrence of spontaneous unprovoked ...

Recording Electrical Activity from Identified Neurons in the Intact Brain of Transgenic Fish

鉴定转基因鱼的完整的大脑中的神经元的电活动记录

Understanding the cell physiology of neural circuits that regulate complex behaviors is greatly enhanced by using model systems in which this work can be ...

Simultaneous Electrophysiological Recording and Calcium Imaging of Suprachiasmatic Nucleus Neurons

Simultaneous electrophysiological and fluorescent imaging recording methods were used to study the role of changes of membrane potential or current in ...

Long-term Behavioral Tracking of Freely Swimming Weakly Electric Fish

自由游泳弱电鱼的长期行为跟踪

Long-term behavioral tracking can capture and quantify natural animal behaviors, including those occurring infrequently. Behaviors such as exploration and ...

Electrophysiological Recording in the Brain of Intact Adult Zebrafish

在完整的成年斑马鱼大脑电生理记录

Previously, electrophysiological studies in adult zebrafish have been limited to slice preparations or to eye cup preparations and electrorentinogram ...

Recording Single Neurons' Action Potentials from Freely Moving Pigeons Across Three Stages of Learning

自由移动鸽子穿越三个阶段学习记录单个神经元&#39;动作电位

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed ...

Extracellular Wire Tetrode Recording in Brain of Freely Walking Insects

外导线四极管记录在行走自如昆虫大脑

Increasing interest in the role of brain activity in insect motor control requires that we be able to monitor neural activity while insects perform ...