Extracellular pH can indicate the variation in organelle function and cell state. It is important to measure extracellular pH (pHe) with a controllable distance. In this work, a potentiometric SECM dual-microelectrode was developed to monitor the pHe of MCF-7 cells under electrical stimulation. The distance between the dual-microelectrode and the cells was determined first with a gold microelectrode by recording the approaching curve, and the pH was determined using an open-circuit potential (OCP) technique with a polyaniline-modified Pt microelectrode. The pH microelectrode showed a response slope of 53.0 ± 0.4 mV/pH and good reversibility from pH 4 to pH 8, fast response within 10 s, and a potential drift of 1.13% for 3 h, and thus was employed to monitor the pHe of stimulated cells. The value of pHe decreased with the decrease in the distance to cells, likely due to the release of H+. With an increase in the stimulation potential or time, the pHe value decreased, as the cell membrane became more permeable, which was verified by fluorescence staining of calcein-AM/PI (propidium iodide). Based on these results, this method can be widely applied for determining the species released by biosystems at a controllable position.
Keywords: Dual-microelectrode; Extracellular pH; Potentiometric sensing; Real-time monitoring; Scanning electrochemical microscopy.