Mootha V K, French S, Balaban R S
Laboratory of Cardiac Energetics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
Anal Biochem. 1996 May 1;236(2):327-30. doi: 10.1006/abio.1996.0174.
Ca(2+)-selective microelectrodes with Simon's neutral carrier ETH 1001 are commercially available and have been widely used for the measurement of both extra- and intracellular calcium. The electrodes demonstrate high selectivity against other cations such as magnesium, sodium, and potassium. We report, however, that the ETH 1001-based microelectrode is a superior tetraphenylphosphonium (TPP+)-sensitive electrode. The electrode exhibits a Nernstian response for [Ca2+] > 10(-5) M but for [TTP+] > 10(-7) M. Using two different methods, we found that log kTTPCa (selectivity coefficient for TPP+ with respect to Ca2+) is in the range of -3.0 to -5.3. We argue that the ETH 1001 microelectrode can be used as a commercially available TPP+ electrode. We illustrate this application by making membrane potential recordings in respiring mitochondria. The results are identical to those obtained using conventional ion-exchange TPP+ electrodes.
带有西蒙中性载体ETH 1001的钙离子选择性微电极在市场上可以买到,并且已被广泛用于细胞外和细胞内钙的测量。这些电极对其他阳离子如镁离子、钠离子和钾离子表现出高选择性。然而,我们报告称,基于ETH 1001的微电极是一种优异的四苯基鏻离子(TPP+)敏感电极。当[Ca2+] > 10(-5) M时,该电极对[Ca2+]呈现能斯特响应,但当[TPP+] > 10(-7) M时也呈现能斯特响应。使用两种不同方法,我们发现log kTTPCa(TPP+相对于Ca2+的选择性系数)在-3.0至-5.3范围内。我们认为ETH 1001微电极可以用作市售的TPP+电极。我们通过在呼吸的线粒体中记录膜电位来说明这种应用。结果与使用传统离子交换TPP+电极获得的结果相同。