Neher E
Methods Enzymol. 1992;207:123-31. doi: 10.1016/0076-6879(92)07008-c.
This chapter describes corrections that have to be applied to measured membrane potentials in patch clamp experiments. Some of them [Eqs. (1)-(3)] are required regardless of the nature of the reference electrode (in the Ringer's solution bath) whenever the pipette-filling solution is different from the bath solution. They represent the liquid junction potentials that are present at the pipette tip before patch formation. In addition, corrections have to be applied when the bath solution is being changed during a measurement (i.e., after seal formation). In that case the following rules apply. (1) The new solution should never get into contact with the bare silver/silver chloride wire of the reference electrode. This requirement is best met by using a salt bridge. (2) The "best" salt bridge is a 3 M KCl bridge with an abrupt KCl-bath fluid boundary at its tip (see above). This bridge does not require any additional potential corrections, but it may lead to KCl poisoning of the bath or become contaminated by solutions used previously. (3) Local solution changes (microperfusion by puffer pipette, U tool or sewer pipe arrangements) as well as recessed KCl bridges require additional corrections, which (together with the simple liquid junction potential correction) are approximately given by Eqs. (6)-(8). It should be stressed that all equations given here represent approximate corrections, since liquid junction potentials are thermodynamically ill-defined. This is particularly relevant for Eqs. (6) and (7) where the sum of two liquid junction potentials appears.
本章描述了在膜片钳实验中对测量的膜电位必须进行的校正。无论参比电极(在林格氏液浴中)的性质如何,只要移液器填充溶液与浴溶液不同,其中一些校正 [式(1)-(3)] 都是必需的。它们代表在形成膜片之前移液器尖端存在的液体接界电位。此外,在测量过程中(即封接形成后)更换浴溶液时也必须进行校正。在这种情况下,适用以下规则。(1) 新溶液绝不应与参比电极的裸银/氯化银导线接触。使用盐桥最能满足这一要求。(2) “最佳”盐桥是一种3 M KCl桥,其尖端有一个陡峭的KCl-浴液边界(见上文)。这种桥不需要任何额外的电位校正,但可能会导致浴液的KCl中毒或被先前使用的溶液污染。(3) 局部溶液变化(通过微量移液器、U形工具或下水道管装置进行微量灌注)以及凹陷的KCl桥需要额外的校正,这些校正(连同简单的液体接界电位校正)大约由式(6)-(8)给出。应该强调的是,这里给出的所有方程都代表近似校正,因为液体接界电位在热力学上定义不明确。这对于式(6)和(7)尤为相关,其中出现了两个液体接界电位的总和。