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. 2019 Dec 6;10:5610. doi: 10.1038/s41467-019-13535-1

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — SICM feedback-controlled real-time pHe detection of living cells. a Bright filed microscopy image showing a pH-sensitive nanoprobe over a buffered single gastric parietal cell (left top). The cartoon at the bottom illustrates the construction of extracellular pH measurements (left bottom). Feedback-control allows for positioning of the pH nanoprobe to accurately detect 100 µM db-cAMP (cAMP analogue) triggered via rapid HCl secretion from a single gastric parietal cell. The secretion can be inhibited with selective H⁺, K^+-ATPase inhibitor SCH28080 (middle). The db-cAMP-induced HCl secretion from the gastric parietal cells was confirmed with pH-sensitive BCECF fluorescent dye by confocal microscopy (right). Changes in fluorescent intensity of pH-sensitive dye can be mapped out either by x–y or by x–z scanning. The dotted crossline marks the Z position in the XZ image and the Y position in the XY image, respectively. b pHe detection from a single low-buffered Coscinodiscus wailesii cell. Changing light conditions from illumination (yellow stripes represent light illumination) to darkness (grey stripes represent darkness), and vice versa revealed rapid pH changes ~1 µm above the cell surface. Change in pH is almost undetectable when the probe is 100 µm away from the cell surface.