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用于测量单个大鼠心肌细胞快速内向电流的膜片钳电压钳方法。

Patch-voltage-clamp method for measuring fast inward current in single rat heart muscle cells.

作者信息

Zilberter Y I, Timin E N, Bendukidze Z A, Burnashev N A

出版信息

Pflugers Arch. 1982 Aug;394(2):150-5. doi: 10.1007/BF00582917.

Abstract

A patch-voltage-clamp method was used to measure fast inward ionic currents in single heart muscle cells. 1. Theoretical analysis including computer simulation has shown that the method provides fast settling of membrane potential (within 10 microseconds) and reliable voltage clamp on the tested membrane patch when the area of the patch is 200-300 times smaller than the area of the whole cell membrane. 2. The transient time at the I-V converter output is increased up to 70-75 microseconds due to the stray capacity in the I-V converter feedback. When fast-response operational amplifiers are used in the set up this transient time may be decreased to 30 microseconds by partial restoration of the high-frequency components of the signal. 3. Experimental data have shown that the ionic channel population in the membrane patch of about 5 micrometers in diameter is on the one hand large enough to directly observe integral ionic current and on the other hand small enough for the fluctuations of ionic current to be appreciable. This permits the method to be applied to ionic current investigations both by classical methods and by statistical analysis.

摘要

采用膜片电压钳方法测量单个心肌细胞中的快速内向离子电流。1. 包括计算机模拟在内的理论分析表明,当膜片面积比整个细胞膜面积小200 - 300倍时,该方法能使膜电位快速稳定(在10微秒内),并在被测膜片上实现可靠的电压钳制。2. 由于I - V转换器反馈中的杂散电容,I - V转换器输出端的瞬态时间增加到70 - 75微秒。当在装置中使用快速响应运算放大器时,通过部分恢复信号的高频成分,该瞬态时间可降至30微秒。3. 实验数据表明,直径约5微米的膜片中的离子通道数量一方面足够大,能够直接观察到整体离子电流,另一方面又足够小,使得离子电流的波动明显。这使得该方法既可以应用于经典方法的离子电流研究,也可以应用于统计分析。

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