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Wiley, Advanced Functional Materials, 12(27), p. 1605533, 2017

DOI: 10.1002/adfm.201605533

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Two-Color Emitting Colloidal Nanocrystals as Single-Particle Ratiometric Probes of Intracellular pH

This paper is made freely available by the publisher.
This paper is made freely available by the publisher.

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Abstract

Intracellular pH is a key parameter in many biological mechanisms and cell metabolism and is used to detect and monitor cancer formation and brain or heart diseases. pH-sensing is typically performed by fluorescence microscopy using pH-responsive dyes. Accuracy is limited by the need for quantifying the absolute emission intensity in living biological samples. An alternative with a higher sensitivity and precision uses probes with a ratiometric response arising from the different pH-sensitivity of two emission channels of a single emitter. Current ratiometric probes are complex constructs suffering from instability and cross-readout due to their broad emission spectra. Here, we overcome such limitations using a single-particle ratiometric pH probe based on dot-in-bulk CdSe/CdS nanocrystals (NCs). These nanostructures feature two fully-separated narrow emissions with different pH sensitivity arising from radiative recombination of core- and shell-localized excitons. The core emission is nearly independent of the pH, whereas the shell luminescence increases in the 3-11 pH range, resulting in a cross-readout-free ratiometric response as strong as 600%. In vitro microscopy demonstrates that the ratiometric response in biologic media resembles the precalibralation curve obtained through far-field titration experiments. The NCs show good biocompatibility, enabling us to monitor in real-time the pH in living cells