Voltage-Clamp and Patch-Clamp Techniques
Voltage-Clamp and Patch-Clamp Techniques
复制标题
电压钳和膜片钳技术
DOI：
10.1007/3-540-26574-0_16
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发表时间：
2005
影响因子：
3.4
通讯作者：
R. Meyer
中科院分区：
文献类型：
--
作者：
H. Polder;M. Weskamp;K. Linz;R. Meyer
本章描述了使用微电极分析可兴奋细胞电行为的各种方法。自20世纪20年代引入以来，微电极已成为电生理学的“主力”，并且存在关于该主题的大量出版物。在这里，我们给出了在单细胞水平上最重要的研究方法的综合概述，然而，高通量方法和自动记录程序没有讨论。本章末尾有一个参考书目（进一步阅读），详细介绍了近几十年来出版的一些最重要的书籍。自世纪以来，人们已经熟知可兴奋细胞能够产生电信号。然而，直到1949年Ling和Gerard发明了第一个玻璃微电极，这些电信号的起源才得以证明。这些玻璃微量移液器首次允许检测细胞的膜电位，因为它们的尖端足够小，可以穿透细胞膜而不破坏细胞，并且它们的高电阻避免了膜电位的分流（参见电极原理一章）。这些微电极连接到电压放大器的高电阻输入端。1951年，发表了第一个使用这些电极在哺乳动物心肌组织上进行的实验（德雷珀和魏德曼1951）。在他们的实验中，可以记录到约-80 mV的负膜电位和心脏动作电位（AP），其典型形状和真实的振幅。电压钳技术（科尔，1949; Hodgkin等，1952）适用于心脏组织（Deck等，1964），揭示了心脏动作电位的不同电流分量，在动作电位期间的膜电流章节中进行了描述。在20世纪70年代后期，分离单个活心肌细胞的技术的发展使心脏生理学的进展又有了一个飞跃（Dow等，1981）。与此同时，发明了膜片钳技术（Hamill et al. 1981）。电极的几何形状的一些小的变化，以及去-
This chapter describes various methods of analysis of the electrical behaviour of excitable cells using microelectrodes. Since their introduction in the 1920’s, microelectrodes have become the “workhorses” of electrophysiology, and a comprehensive number of publications exist about this topic. Here, we give a consolidated overview of the most important methods of investigation at single cell level, however, high throughput methods and automated recording procedures are not discussed. There is a reference list (Further Reading) at the end of this chapter, detailing some of the most important books published in recent decades. Since the nineteenth century, it has been well known that excitable cells are able to produce electrical signals. However, until the invention of the first glass microelectrodes, by Ling and Gerard in 1949, the origin of these electrical signals could not be proven. For the first time, these glass micropipettes allowed the detection of the membrane potential of a cell, as their tips were small enough to penetrate the cell membrane without destroying the cells, and their high electrical resistance avoided shunting the membrane potential (see chapter Principles of Electrodes). These microelectrodes were connected to the high resistance input of a voltage amplifier. The monitoring of the membrane potential, delivered the base of our actual understanding of electrical events inside the heart.In 1951, the first experiments on mammalian heart muscle tissue using these electrodes were published (Draper and Weidmann 1951). During their experiments, the negative membrane potential of around–80 mV and the cardiac action potential (AP), in its typical shape and with its real amplitude, could be recorded. Application of the voltage clamp technique (Cole 1949; Hodgkin et al. 1952), adapted to cardiac tissue (Deck et al. 1964), revealed the different current components of the cardiac action potential, which are described in chapter Membrane Currents During the Action Potential. A further leap in progress for cardiac physiology was gained by the development of techniques to isolate single living cardiac myocytes in the late 1970’s (Dow et al. 1981). At the same time, the patch clamp technique was invented (Hamill et al. 1981). Some small changes in the geometry of the electrodes, as well as the de-