Name: fabrication of amperometric electrodes
Uploaded: 2009-05-04T00:00:00
Description: Watch this Scientific Journal Video about Fabrication of Amperometric Electrodes at JoVE.com

<p class="jove_content">我们感谢阿龙体育福克斯博士在芝加哥大学，在他的实验室，这一协议最初是设计，和威廉罗登（圣路易斯大学）协议的进一步发展。 </p><p class="jove_content">我们感谢白厅基金会和美国国家科学基金会的支持。 </p>

<p class="jove_title">第1部分：碳纤维的制备</p><ol><li>切割长度〜1.25 X的毛细吸管长度的碳纤维束（使用7微米直径）。 </li><li>煮沸删除上浆〜100毫升的容器中的丙酮，并添加纤维热，煮沸30分钟或更长的时间丙酮。 </li><li>删除从温暖丙酮的纤维，并转移到一个干净的烧杯50-100毫升新鲜丙酮。 </li><li>放置在一个开放的150毫米培养皿的捆绑和airdry通宵干纤维。 </li></ol><p class="jove_title">第2部分：消防抛光玻璃毛细管</p><ol><l...

<ol>
	<li>Mundorf, M. L., Wightman, 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=Amperometry+and+cyclic+voltammetry+with+carbon+fiber+microelectrodes+at+single+cells.">Amperometry and cyclic voltammetry with carbon fiber microelectrodes at single cells.</a> <em style='font-style: italic'>Curr Protocols in Neurosci</em>. <b>6</b> (14), 1-6 (2002).</li><li>Machado, D. J., Montesinos, M. S., Borges, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Good+practices+in+single-cell+amperometry.">Good practices in single-cell amperometry.</a> <em style='font-style: italic'>Methods in Molec. Biol</em>. <b>440</b>, 297-313 (2008).</li></ol>

<p class="jove_content">本协议描述了如何生成和减少安培的电极。安培录音期间，碳纤维电极放在对一个分泌细胞的表面。 Exocytotic活动是观察安培的峰值，这表明的儿茶酚胺的氧化后的电子转移所造成的电化学电流。与实践和切割安培电极，可大大改善噪音水平超过许多市售电极。 </p>

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Epoxy Hardener 1,3-phenylene-diamene flakes</td>
<td>Reagent</td>
<td>Sigma Aldrich</td>
<td>P23954-100G04 716AJ</td>
<td>Aliquots are stored in a dessicator and should not be exposed to light.</td>
</tr>
<tr>
<td>Carbon Fibers</td>
<td>Material</td>
<td>Goodfellow</td>
<td>C005725</td>
<td>&nbsp;</td>
<tr>
<td>Micro-Hematocrit Capillary Tubes</td>
<td>Material</td>
<td>Fisher</td>
<td>NC9836768</td>
<td>&nbsp;</td>
<tr>
<td>Epoxy Resin</td>
<td>Reagent</td>
<td>Miller-Stephenson</td>
<td>Epon Resin 828</td>
<td>&nbsp;</td>
<tr>
<td>Dissecting Microscope</td>
<td>Instrument</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Electrode Puller</td>
<td>Instrument</td>
<td>Narishige Japan</td>
<td>PP-830</td>
<td>1st pull = 24.8 degrees C; 2nd pull = 62.7 degrees C</td>
</tr>
<tr>
<td>Oven</td>
<td>Instrument</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>set to 100 degrees F.</td>
</tr>
<tr>
<td>Hot plate</td>
<td>Instrument</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>set to 55 degrees C</td>
</tr>		</tbody>
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fabrication of amperometric electrodes

<p class="jove_content">碳纤维电极安培测量单细胞囊泡检测儿茶酚胺释放的关键。在这里，我们描述产生低噪声（&lt;0.5 PA）电极所需的技术。从公布的描述，该技术已被修改以前的研究（1,2）。电极是由准备碳纤维和线程到每个毛细管单独使用一个过滤器，真空吸纤维。下一步，与纤维的毛细管被拉到一个电极拉马，创建两半，每一个细尖。电极浸在热，液体环氧树脂与固化剂混合，以创建一个环氧玻璃密封。最后，电极置于烘箱中固化环氧树脂。电极小心处理，以确保它们是一致和无损伤的关键。此协议显示了如何从单细胞记录安培的电极制造和切割。</p>

Watch this Scientific Journal Video about Fabrication of Amperometric Electrodes at JoVE.com

Fabrication of Amperometric Electrodes

<p class="jove_content">本协议描述了如何生成的碳纤维电极。随后，电极用于检测儿茶酚胺释放囊泡与碳纤维安培。</p>

安培电极的制备

Carbon fiber electrodes are crucial for the detection of catecholamine release from vesicles in single cells for amperometry measurements.  Here, we ...

Research

JoVE Journal

Biology

Application of Light-cured Dental Adhesive Resin for Mounting Electrodes or Microdialysis Probes in Chronic Experiments

光固化牙科粘合树脂安装电极或微透析探针在慢性实验中的应用

In chronic recording experiments, self-curing dental acrylic resins have been used as a mounting base of electrodes or microdialysis-probes. Since these ...

科研

生物学

Patterning Cells on Optically Transparent Indium Tin Oxide Electrodes

在光学透明的铟锡氧化物电极的图案细胞

The ability to exercise precise spatial and temporal control over cell-surface interactions is an important prerequisite to the assembly of multi-cellular ...

Fabrication of a Microfluidic Device for the Compartmentalization of Neuron Soma and Axons

条块索玛的神经元和轴突的一个微流体设备制造

In this video, we demonstrate the technique of soft lithography with polydimethyl siloxane (PDMS) which we use to fabricate a microfluidic device for ...

PDMS Device Fabrication and Surface Modification

PDMS的设备制造和表面改性

Non-plasma Bonding of PDMS for Inexpensive Fabrication of Microfluidic Devices

PDMS微流体装置的廉价制造的非等离子键合

In this video, we demonstrate how to use the neuron microfluidic device without plasma bonding.  In some cases it may be desirable to reversibly bond ...

Fabrication of the Thermoplastic Microfluidic Channels

制备热塑性微流体通道

In our lab, we have successfully isolated nucleic acids directly from microliter and submicroliter volumes of human blood, urine and stool using ...

Making Patch-pipettes and Sharp Electrodes with a Programmable Puller

膜片吸液管和夏普电极与一个可编程的拖轮

Glass microelectrodes (also called pipettes) have been a workhorse of electrophysiology for decades.  Today, such pipettes are made from glass capillaries ...

Micro-drive Array for Chronic <em>in vivo</em> Recording: Drive Fabrication

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

慢性的微型驱动器阵列<em>在体内</em>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. ...

Fabrication of Myogenic Engineered Tissue Constructs

制作生肌工程的组织结构

Despite the fact that electronic pacemakers are life-saving medical devices, their long-term performance in pediatric patients can be problematic owing to ...

Fabrication and Operation of an Oxygen Insert for Adherent Cellular Cultures 

为贴壁细胞培养的氧气插入的制备及操作

Oxygen is a key modulator of many cellular pathways, but current devices permitting in vitro oxygen modulation fail to meet the needs of biomedical ...