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Super-resolution imaging of non-fluorescent reactions via competition

Abstract

Super-resolved fluorescence microscopy techniques have enabled substantial advances in the chemical and biological sciences. However, they can only interrogate entities that fluoresce, and most chemical or biological processes do not involve fluorescent species. Here we report a competition-enabled imaging technique with super-resolution (COMPEITS) that enables quantitative super-resolution imaging of non-fluorescent processes. It is based on the incorporation of competition into a single-molecule fluorescence-detection scheme. We demonstrate COMPEITS by investigating a photoelectrocatalytic reaction; we map, with nanometre precision, a non-fluorescent surface reaction that is important for water decontamination on single photocatalyst particles. The subparticle-level quantitative information of reactant adsorption affinities unambiguously decouples size- and shape-scaling laws on specific particle facets and uncovers a surprising biphasic shape dependence, leading to catalyst design principles for optimal reactant adsorption efficacy. With its ability to provide spatially resolved information on the behaviours of unlabelled, non-fluorescent entities under operando conditions, COMPEITS could interrogate a variety of surface processes in fields ranging from heterogeneous catalysis and materials engineering to nanotechnology and energy sciences.

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Data availability

The data that support the findings of this study are available from the corresponding author on reasonable request.

Code availability

The MATLAB codes that support the findings of this study are available from the corresponding author on reasonable request.

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Acknowledgements

The research on water pollutant degradation and super-resolution mapping was funded by the Army Research Office (grant no. W911NF-17-1-0590, and in part grant no. W911NF-18-1-0217). The research on BiVO4 synthesis, characterization and photoelectrochemistry was funded by the US Department of Energy, Office of Science, Basic Energy Sciences, Catalysis Science Program (grant no. DE-SC0004911). This research made use of the Cornell Center for Materials Research Shared Facilities, which are supported through the NSF MRSEC program (grant no. DMR-1719875). We thank W. Jung and G. Chen for helpful discussions on instrument construction and data analysis.

Author information

X.M., C.L. and P.C. designed the research. X.M. synthesized the catalysts, constructed the instruments, performed the measurements, coded the software and analysed the data. X.M. and P.C. discussed the results and wrote the manuscript. M.H. and N.Z. contributed to the experiments.

Correspondence to Peng Chen.

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Competing interests

X.M. and P.C. have filed a provisional patent application ‘A competition-enabled imaging technique with super-resolution’ on 27 September 2018 with the US Patent Office (Provision Number 62/737,195).

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Supplementary information

Supplementary Information

Supplementary materials and methods, Supplementary data and discussions, Supplementary Figs. 1–19, Supplementary Tables 1–7

Supplementary Video 1

A representative fluorescence video segment of the BiVO4 particle shown in Fig. 2 in the presence of 50 nM AR without HQ, at a frame rate of 133 frames per second (about twice the actual frame rate). Video frame dimensions: 10.1 × 12.5 μm2 (that is, 38 × 47 pixel2, in which each pixel is 266 × 266 nm2). Video length: 7,000 frames, which corresponds to an actual experimental time of 105 seconds. ImageJ is the preferred program for viewing this video at optimal quality. This video shows that there are stochastic appearances of the fluorescent product molecule resorufin from the auxiliary fluorogenic reaction of AR oxidation (that is, substantially brighter spots than the photoluminescence background of the BiVO4 particle) and those bright spots appear more frequently on the lateral facet of the particle than on its basal facet. A representative single-frame fluorescence image is shown in Supplementary Fig. 6.

Supplementary Video 2

A representative fluorescence video segment of the BiVO4 particle shown in Fig. 2 in the presence of 50 nM AR with 250 µM HQ, at a frame rate of 133 frames per second (about twice the actual frame rate). Video frame dimensions: 10.1 × 12.5 μm2 (that is, 38 × 47 pixel2, in which each pixel is 266 × 266 nm2). Video length: 7,000 frames, which corresponds to an actual experimental time of 105 seconds. ImageJ is the preferred program for viewing this video at optimal quality. This video shows that there are stochastic appearances of the fluorescent product molecule resorufin from the auxiliary fluorogenic reaction of AR oxidation (that is, substantially brighter spots than the photoluminescence background of the BiVO4 particle) and those bright spots appear more frequently on the lateral facet of the particle than on its basal facet. In the presence of the competitor HQ, the overall appearance frequency of the fluorescent product molecule decreases. A representative single-frame fluorescence image is shown in Supplementary Fig. 6.

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Figure 1: A demonstration of COMPEITS using a surface reaction.
Figure 2: COMPEITS imaging of the photoelectrooxidation of HQ on single BiVO4 particles.
Figure 3: Subparticle, facet-specific size and shape dependences of HQ binding affinity.
Figure 4: Rational design of size- and shape-tunable BiVO4 particles for optimal reactant adsorption.