<p>从RW Aldrich公司的实验室解决方案处方萎缩。我们感谢以下资助机构的支持和基础：美国国立卫生研究院，唐纳德B.肌肉萎缩症协会，美国国家科学基金会，美国心脏协会，和迪莉娅E.巴克斯特基金会，Klingenstein基金和神经科学的麦克奈特基金会。 </p>

<p class="jove_step">第1部分：删除卵黄膜</p><ol><li>准备两套镊子。我们更喜欢5号钳，其中一组已略微削尖文件。修整和抛光火7“巴斯德吸管，还要准备一个玻璃移液管。 </li><li> <em>建议：</em>为了防止打滑的卵母细胞，切圈电网（1毫米广场）和胶成一个60毫米培养皿的底部。此菜可重复使用，无限期如果使用之间保持清洁。 </li><li>高渗溶液（配方），在解剖显微镜下填写的培养皿一半。 </li><li>取出1-3爪蟾卵母细胞，...

<ol>
	<li>Neher, E., Sakmann, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single-channel+currents+recorded+from+membrane+of+denervated+frog+muscle+fibres.">Single-channel currents recorded from membrane of denervated frog muscle fibres.</a> <em style='font-style: italic'>Nature</em>. <b>260</b>, 799-802 (1976).</li><li>Sackmann, B., Neher, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> <em style='font-style: italic'>Single-channel Recording</em>. , (1983).</li></ol>

<p>有许多的电生理记录参数，这里不讨论。钻机安装，系统噪声管理，渠道表达，和记录协议都良好的实验结果也很关键。 </p><p>根据我们的经验，这些都是形成密封可靠，最关键的参数：高品质的卵母细胞，迅速取出卵黄膜（又名“剥皮”），使用新拉的移液器防尘，流畅的移液器，积极施加压力，在入浴时解决方案，在洗澡的简短的时间之前密封。话虽如此，经验差别很大，别人有自己的一组最重要的因素。 </p>

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<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>Patch Clamp Amplifier & Software</td>
<td>Instrument</td>
<td>HEKA or Axon</td>
<td>EPC-10</td>
<td>Or other similar device (e.g. HEKA EPC-9 or Axopatch 200B)</td>
</tr>
<tr>
<td>No. 5 forceps (two pairs)</td>
<td>Tool</td>
<td>Fine Science Tools</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Oocyte shrinking solution</td>
<td>Reagent</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Ingredient                            In mM
N-methyl-D-glucamine     220
HEPES                                   10
MgCl2                                       1
EGTA                                       10
Aspartic Acid                        220
KCl                                             2
pH to 7.4 w/ N-methyl-D-glucamine</td>
</tr>
<tr>
<td>woven mesh</td>
<td>800 um</td>
<td>Spectrum Laboratories INC</td>
<td>146481</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>
	

patch clamp recording of ion channels expressed in  xenopus oocytes

<p class="jove_content">由于其发展索克曼和内尔<sup>1，2，</sup>膜片钳已成为一个极为有用的方法，为单个或多个细胞离子通道的电生理测量。这种技术可以应用到离子通道，在其原生环境，并在异源细胞，如从非洲爪蛙，非洲爪蟾收获的卵母细胞，表达的。在这里，我们描述了补丁钳爪蟾卵母细胞的行之有效的技术。这项技术是用来衡量人口（macropatch）或单独（单声道录音）的表达离子通道的特性。我们专注于技术，以最大限度地提高卵母细胞的制备和密封代质量。随着各方面的因素进行了优化，这种技术提供了一个成功的印章代概率在90％以上。基于他或她认为最重要的因素的每一个研究员，这个过程可能是不同的优化，我们目前的方法，导致在我们手中最大的成功。 </p>

Watch this Scientific Journal Video about Patch Clamp Recording of Ion Channels Expressed in  Xenopus Oocytes at JoVE.com

Patch Clamp Recording of Ion Channels Expressed in  Xenopus Oocytes

<p class="jove_content">这是为了介绍，从非洲爪蟾卵母细胞来修补钳记录。它涵盖了卵黄膜去除，形成一个gigaohm密封（gigaseal），和可选的补丁外拓扑的转换。</p>

补丁钳爪蟾卵母细胞中表达的离子通道

Since its development by Sakmann and Neher 1, 2, the patch clamp has become established as an extremely useful technique for electrophysiological ...

Research

JoVE Journal

Biology

Methods for Patch Clamp Capacitance Recordings from the Calyx

膜片钳从花萼电容录音方法

We demonstrate the basic techniques for presynaptic patch clamp recording at the calyx of Held, a mammalian central nervous system nerve terminal. ...

科研

生物学

Pressure-polishing Pipettes for Improved Patch-clamp Recording

改进的膜片钳记录的压力抛光移液器

Pressure-polishing is a method for shaping glass pipettes for patch-clamp recording.  We first developed this method for fabricating pipettes suitable for ...

Microinjection of Xenopus Laevis Oocytes

非洲爪蟾卵母细胞的微量注射

Microinjection of Xenopus laevis oocytes followed by thin-sectioning electron microscopy (EM) is an excellent system for studying nucleocytoplasmic ...

Visualizing RNA Localization in <em>Xenopus</em> Oocytes

Visualizing RNA Localization in Xenopus Oocytes

可视化的RNA中的本地化<em>爪蟾</em>卵母细胞

RNA localization is a conserved mechanism of establishing cell polarity.  Vg1 mRNA localizes to the vegetal pole of Xenopus laevis oocytes and acts to ...

Fertilization of <em>Xenopus oocytes</em> using the Host Transfer Method

Fertilization of Xenopus oocytes using the Host Transfer Method

施肥<em>爪蟾卵母细胞</em>使用主机传输方法

Studying the contribution of maternally inherited molecules to vertebrate early development is often hampered by the time and expense necessary to ...

Mutagenesis and Functional Analysis of Ion Channels Heterologously Expressed in Mammalian Cells

异源表达在哺乳动物细胞离子通道的突变和功能分析

We will demonstrate how to study the functional effects of introducing a point mutation in an ion channel. We study G protein-gated inwardly rectifying ...

Patch Clamp and Perfusion Techniques for Studying Ion Channels Expressed in <em>Xenopus</em> oocytes

Patch Clamp and Perfusion Techniques for Studying Ion Channels Expressed in Xenopus oocytes

膜片钳和灌注技术研究离子通道的表达<em>爪蟾</em>卵母细胞

The protocol presented here is designed to study the activation of the large conductance, voltage- and Ca2+-activated K+ (BK) channels. The protocol may ...

Optimized Transfection Strategy for Expression and Electrophysiological Recording of Recombinant Voltage-Gated Ion Channels in HEK-293T Cells

优化转染的HEK - 293T细胞中的表达和重组的电压门控离子通道的电生理记录策略

The in vitro expression and electrophysiological recording of recombinant voltage-gated ion channels in cultured human embryonic kidney cells (HEK-293T) ...

Giant Liposome Preparation for Imaging and Patch-Clamp Electrophysiology

巨脂质体制剂成像和膜片钳电

The reconstitution of ion channels into chemically defined lipid membranes for electrophysiological recording has been a powerful technique to identify ...

The <em>Xenopus</em> Oocyte Cut-open Vaseline Gap Voltage-clamp Technique With Fluorometry

The Xenopus Oocyte Cut-open Vaseline Gap Voltage-clamp Technique With Fluorometry

该<em&gt;爪蟾</em&gt;卵母细胞切割开凡士林隙电压钳技术与荧光

The cut-open oocyte Vaseline gap (COVG) voltage clamp technique allows for analysis of electrophysiological and kinetic properties of heterologous ion ...