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Supercharging: a method for improving patch-clamp performance.

机译:增压:一种改善膜片钳性能的方法。

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摘要

Patch-clamp performance can be improved without altering the normal headstage configuration described by (Hamill, O. P., A. Marty, E. Neher, B. Sakmann, and F. J. Sigworth, 1981, Pfluegers Arch. Eur. J. Physiol., 391:85-100). The "supercharging" method permits resolution of such fast events as calcium and sodium tail currents. Digital computer modeling and analog electronic simulation were used to identify appropriate shapes for the command voltage and the voltage applied to a capacitor tied to the input of the headstage. The voltage command pulse consists of a step with a brief (5-15 microseconds) rectangular spike on its leading edge. Spike amplitude is a function of the membrane capacitance and the access resistance. The spike drives current through the access resistance and speeds charging of the membrane capacitance, making it possible to complete a voltage step within 5-15 microseconds. Clamping speed is independent of the electrode and feedback resistance over a wide range. The second function of the patch clamp amplifier is current measurement, and good time resolution requires suppression of the capacity transient. This can be accomplished by applying an appropriately shaped voltage to the small capacitor tied to the input of the headstage. Series resistance compensation for ionic current transients does not interfere with supercharging. Although the focus of this paper is on whole cell recording, the supercharging concept may prove useful for single channel and bilayer recording techniques.
机译:膜片钳性能可在不改变(Hamill,OP,A.Marty,E.Neher,B.Sakmann,and FJ Sigworth,1981,Pfluegers Arch.Eur.J.Physiol。,391: 85-100)。 “增压”方法可以解决诸如钙和钠尾电流之类的快速事件。使用数字计算机建模和模拟电子仿真来识别命令电压的适当形状,以及施加到与前置输入相连的电容器上的电压。电压指令脉冲由一个台阶组成,该台阶在其前沿具有一个短暂的(5-15微秒)矩形尖峰。尖峰幅度是膜电容和访问电阻的函数。尖峰驱动电流通过访问电阻并加快膜电容的充电速度,从而有可能在5到15微秒内完成电压阶跃。在很宽的范围内,夹紧速度与电极和反馈电阻无关。膜片钳放大器的第二个功能是电流测量,良好的时间分辨率要求抑制电容瞬变。这可以通过将适当形状的电压施加到与前置输入相连的小电容器上来实现。离子电流瞬变的串联电阻补偿不会干扰增压。尽管本文的重点是全单元记录,但是增压概念可能对单通道和双层记录技术很有用。

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