Abstract
Multiplexed intracellular detection is desirable in biomedical sciences for its higher efficiency and accuracy compared to the single-analyte detection. However, it is very challenging to construct nanoprobes that possess multiple fluorescent signals to recognize the different intracellular species synchronously. Herein, we proposed a novel dual-excitation/dual-emission upconversion strategy for multiplexed detection through the design of upconversion nanoparticles (UCNP) loaded with two dyes for sensitization and quenching of the upconversion luminescence (UCL), respectively. Based on the two independent energy transfer processes of near-infrared (NIR) dye IR845 to UCNP and UCNP to visible dye PAPS-Zn, ClO− and Zn2+ were simultaneously detected with a limit of detection (LOD) of 41.4 and 10.5 nM, respectively. By utilizing a purpose-built 830/980 nm dual-laser confocal microscope, both intrinsic and exogenous ClO− and Zn2+ in live MCF-7 cells have been accurately quantified. Such dual-excitation/dual-emission ratiometric UCL detection mode enables not only monitoring multiple intracellular analytes but also eliminating the detection deviation caused by inhomogeneous probe distribution in cells. Through modulation of NIR dye and visible dye with other reactive groups, the nanoprobes can be extended to analyze various intracellular species, which provides a promising tool to study the biological activities in live cells and diagnose diseases.
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Kolanowski, J. L.; Liu, F.; New, E. J. Fluorescent probes for the simultaneous detection of multiple analytes in biology. Chem. Soc. Rev.2018, 47, 195–208.
Fan, Y.; Wang, S. F.; Zhang, F. Optical multiplexed bioassays for improved biomedical diagnostics. Angew. Chem., Int. Ed.2019, 58, 13208–13219.
Leng, Y.; Sun, K.; Chen, X. Y.; Li, W. W. Suspension arrays based on nanoparticle-encoded microspheres for high-throughput multiplexed detection. Chem. Soc. Rev.2015, 44, 5552–5595.
Jia, P. P.; Jiang, S. T.; Xu, L. Small-molecule bifunctional fluorescent probes for the differential detection of multiple guests. Curr. Org. Synth.2019, 16, 485–497.
Zhou, W. J.; Liang, W. B.; Li, D. X.; Yuan, R.; Xiang, Y. Dual-color encoded DNAzyme nanostructures for multiplexed detection of intracellular metal ions in living cells. Biosens. Bioelectron.2016, 85, 573–579.
Mimitou, E. P.; Cheng, A.; Montalbano, A.; Hao, S.; Stoeckius, M.; Legut, M.; Roush, T.; Herrera, A.; Papalexi, E.; Ouyang, Z. Q. et al. Multiplexed detection of proteins, transcriptomes, clonotypes and CRISPR perturbations in single cells. Nat. Methods2019, 16, 409–412.
Haase, M.; Schäfer, H. Upconverting nanoparticles. Angew. Chem., Int. Ed.2011, 50, 5808–5829.
Gu, Z. J.; Yan, L.; Tian, G.; Li, S. J.; Chai, Z. F.; Zhao, Y. L. Recent advances in design and fabrication of upconversion nanoparticles and their safe theranostic applications. Adv. Mater.2013, 25, 3758–3779.
Zhao, L. Z.; Peng, J. J.; Huang, Q.; Li, C. Y.; Chen, M.; Sun, Y.; Lin, Q. N.; Zhu, L. Y.; Li, F. Y. Near-infrared photoregulated drug release in living tumor tissue via yolk-shell upconversion nanocages. Adv. Funct. Mater.2014, 24, 363–371.
Zheng, W.; Huang, P.; Tu, D. T.; Ma, E.; Zhu, H. M.; Chen, X. Y. Lanthanide-doped upconversion nano-bioprobes: Electronic structures, optical properties, and biodetection. Chem. Soc. Rev.2015, 44, 1379–1415.
Li, X. M.; Zhang, F.; Zhao, D. Y. Lab on upconversion nanoparticles: Optical properties and applications engineering via designed nanostructure. Chem. Soc. Rev.2015, 44, 1346–1378.
Yu, S. H.; Tu, D. T.; Lian, W.; Xu, J.; Chen, X. Y. Lanthanide-doped near-infrared II luminescent nanoprobes for bioapplications. Sci. China Mater.2019, 62, 1071–1086.
Xu, J. T.; Yang, P. P.; Sun, M. D.; Bi, H. T.; Liu, B.; Yang, D.; Gai, S. L.; He, F.; Lin, J. Highly emissive dye-sensitized upconversion nanostructure for dual-photosensitizer photodynamic therapy and bioimaging. ACS Nano2017, 11, 4133–4144.
Liu, J.; Bu, W. B.; Pan, L. M.; Shi, J. L. NIR-triggered anticancer drug delivery by upconverting nanoparticles with integrated azobenzene-modified mesoporous silica. Angew. Chem., Int. Ed.2013, 52, 4375–4379.
Gu, B.; Zhang, Q. C. Recent advances on functionalized upconversion nanoparticles for detection of small molecules and ions in biosystems. Adv. Sci.2018, 5, 1700609.
Li, Z. H.; Yuan, H.; Yuan, W.; Su, Q. Q.; Li, F. Y. Upconversion nanoprobes for biodetections. Coord. Chem. Rev.2018, 354, 155–168.
Peng, J. J.; Xu, W.; Teoh, C. L.; Han, S. Y.; Kim, B.; Samanta, A.; Er, J. C.; Wang, L.; Yuan, L.; Liu, X. G. et al. High-efficiency in vitro and in vivo detection of Zn2+ by dye-assembled upconversion nanoparticles. J. Am. Chem. Soc.2015, 137, 2336–2342.
Zhou, Y.; Chen, W. Q.; Zhu, J. X.; Pei, W. B.; Wang, C. Y.; Huang, L.; Yao, C.; Yan, Q. Y.; Huang, W.; Loo, J. S. C. et al. Inorganic-organic hybrid nanoprobe for NIR-excited imaging of hydrogen sulfide in cell cultures and inflammation in a mouse model. Small2014, 10, 4874–4885.
Hu, J. S.; Wang, R. N.; Fan, R. R.; Huang, Z. H.; Liu, Y. X.; Guo, G.; Fu, H. Enhanced luminescence in Yb3+ doped core-shell upconversion nanoparticles for sensitive doxorubicin detection. J. Lumin.2020, 217, 116812.
Mou, X. M.; Wang, J. X.; Meng, X. F.; Liu, J. L.; Shi, L. Y.; Sun, L. N. Multifunctional nanoprobe based on upconversion nanoparticles for luminescent sensing and magnetic resonance imaging. J. Lumin.2017, 190, 16–22.
Ai, X. Z.; Wang, Z. M.; Cheong, H.; Wang, Y.; Zhang, R. C.; Lin, J.; Zheng, Y. J.; Gao, M. Y.; Xing, B. G. Multispectral optoacoustic imaging of dynamic redox correlation and pathophysiological progression utilizing upconversion nanoprobes. Nat. Commun.2019, 10, 1087.
Zheng, J. D.; Wu, Y. X.; Xing, D.; Zhang, T. Synchronous detection of glutathione/hydrogen peroxide for monitoring redox status in vivo with a ratiometric upconverting nanoprobe. Nano Res.2019, 12, 931–938.
Ke, J. X.; Lu, S.; Shang, X. Y.; Liu, Y.; Guo, H. H.; You, W. W.; Li, X. J.; Xu, J.; Li, R. F.; Chen, Z. et al. A strategy of NIR dual-excitation upconversion for ratiometric intracellular detection. Adv. Sci.2019, 6, 1901874.
Zhou, Y.; Pei, W. B.; Wang, C. Y.; Zhu, J. X.; Wu, J. S.; Yan, Q. Y.; Huang, L.; Huang, W.; Yao, C.; Loo, J. S. C. et al. Rhodaminemodified upconversion nanophosphors for ratiometric detection of hypochlorous acid in aqueous solution and living cells. Small2014, 10, 3560–3567.
Chang, C. J.; Jaworski, J.; Nolan, E. M.; Sheng, M.; Lippard, S. J. A tautomeric zinc sensor for ratiometric fluorescence imaging: Application to nitric oxide-induced release of intracellular zinc. Proc. Natl. Acad. Sci. USA2004, 101, 1129–1134.
Matsui, H.; Oyama, T. M.; Okano, Y.; Hashimoto, E.; Kawanai, T.; Oyama, Y. Low micromolar zinc exerts cytotoxic action under H2O2-induced oxidative stress: Excessive increase in intracellular Zn2+ concentration. Toxicology2010, 276, 27–32.
Hu, P.; Wang, R.; Zhou, L.; Chen, L.; Wu, Q. S.; Han, M. Y.; El-Toni, A. M.; Zhao, D. Y.; Zhang, F. Near-infrared-activated upconversion nanoprobes for sensitive endogenous Zn2+ detection and selective on-demand photodynamic therapy. Anal. Chem.2017, 89, 3492–3500.
You, W. W.; Tu, D. T.; Zheng, W.; Shang, X. Y.; Song, X. R.; Zhou, S. Y.; Liu, Y.; Li, R. F.; Chen, X. Y. Large-scale synthesis of uniform lanthanide-doped NaREF4 upconversion/downshifting nanoprobes for bioapplications. Nanoscale2018, 10, 11477–11484.
Wu, X.; Lee, H.; Bilsel, O.; Zhang, Y. W.; Li, Z. J.; Chen, T.; Liu, Y.; Duan, C. Y.; Shen, J.; Punjabi, A. et al. Tailoring dye-sensitized upconversion nanoparticle excitation bands towards excitation wavelength selective imaging. Nanoscale2015, 7, 18424–18428.
Bogdan, N.; Vetrone, F.; Ozin, G. A.; Capobianco, J. A. Synthesis of ligand-free colloidally stable water dispersible brightly luminescent lanthanide-doped upconverting nanoparticles. Nano Lett.2011, 11, 835–840.
Li, H. H.; Guan, L. M.; Zhang, X. J.; Yu, H.; Huang, D. J.; Sun, M. T.; Wang, S. H. A cyanine-based near-infrared fluorescent probe for highly sensitive and selective detection of hypochlorous acid and bioimaging. Talanta2016, 161, 592–598.
Wang, J. Y.; Niu, Y. M.; Zhang, C.; Chen, Y. Q. A micro-plate colorimetric assay for rapid determination of trace zinc in animal feed, pet food and drinking water by ion masking and statistical partitioning correction. Food Chem.2018, 245, 337–345.
Chen, G. Y.; Damasco, J.; Qiu, H. L.; Shao, W.; Ohulchanskyy, T. Y.; Valiev, R. R.; Wu, X.; Han, G.; Wang, Y.; Yang, C. H. et al. Energy-cascaded upconversion in an organic dye-sensitized core/shell fluoride nanocrystal. Nano Lett.2015, 15, 7400–7407.
Wu, X.; Zhang, Y. W.; Takle, K.; Bilsel, O.; Li, Z. J.; Lee, H.; Zhang, Z. J.; Li, D. S.; Fan, W.; Duan, C. Y. et al. Dye-sensitized core/active shell upconversion nanoparticles for optogenetics and bioimaging applications. ACS Nano2016, 10, 1060–1066.
Dong, H.; Du, S. R.; Zheng, X. Y.; Lyu, G. M.; Sun, L. D.; Li, L. D.; Zhang, P. Z.; Zhang, C.; Yan, C. H. Lanthanide nanoparticles: From design toward bioimaging and therapy. Chem. Rev.2015, 115, 10725–10815.
Acknowledgements
This work was supported by the Science and Technology Cooperation Fund between Chinese and Australian Governments (No. 2017YFE0132300), the Strategic Priority Research Program of the CAS (No. XDB20000000), the National Natural Science Foundation of China (No. 51672272, 21771185, 21771178, and 21975257), Youth Innovation Promotion Association of CAS (No. 2017347), and the CAS/SAFEA International Partnership Program for Creative Research Teams.
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Ke, J., Lu, S., Li, Z. et al. Multiplexed intracellular detection based on dual-excitation/dual-emission upconversion nanoprobes. Nano Res. 13, 1955–1961 (2020). https://doi.org/10.1007/s12274-020-2837-2
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DOI: https://doi.org/10.1007/s12274-020-2837-2