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Single europium-doped nanoparticles measure temporal pattern of reactive oxygen species production inside cells

Nature Nanotechnology volume 4, pages 581–585 (2009)Cite this article

Abstract

Low concentrations of reactive oxygen species, notably hydrogen peroxide (H2O2), mediate various signalling processes in the cell1,2. Production of these signals is highly regulated3 and a suitable probe is needed to measure these events. Here, we show that a probe based on a single nanoparticle can quantitatively measure transient H2O2 generation in living cells. The Y0.6Eu0.4VO4 nanoparticles undergo photoreduction under laser irradiation but re-oxidize in the presence of oxidants, leading to a recovery in luminescence. Our probe can be regenerated and reliably detects intracellular H2O2 with a 30-s temporal resolution and a dynamic range of 1–45 µM. The differences in the timing of intracellular H2O2 production triggered by different signals were also measured using these nanoparticles. Although the probe is not selective towards H2O2, in many signalling processes H2O2 is, however, the dominant oxidant3,4,5,6. In conjunction with appropriate controls, this probe is a powerful tool for unravelling pathways that involve reactive oxygen species.

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Figure 1: Luminescence properties of silica-coated Y0.6Eu0.4VO4 nanoparticles spin-coated on silica coverslips.
Figure 2: Quantitative detection of H2O2.
Figure 3: Intracellular H2O2 produced by VSMCs after ET-1 stimulation.
Figure 4: Intracellular H2O2 produced after PDGF stimulation.

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Acknowledgements

We thank G. Mialon and M. Moreau for nanoparticle synthesis.

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Author notes

  1. Didier Casanova, Cédric Bouzigues and Thanh-Liêm Nguyên: These authors contributed equally to this work

Authors and Affiliations

  1. Laboratoire d'Optique et Biosciences, Ecole Polytechnique, CNRS, INSERM U696, Palaiseau, 91128, France

    Didier Casanova, Cédric Bouzigues, Thanh-Liêm Nguyên, Rivo O. Ramodiharilafy, Latifa Bouzhir-Sima & Antigoni Alexandrou

  2. Laboratoire de Physique de la Matière Condensée, Ecole Polytechnique, CNRS, Palaiseau, 91128, France

    Thierry Gacoin & Jean-Pierre Boilot

  3. Paris-Cardiovascular Research Centre, INSERM U970, 56 rue Leblanc, Paris, 75015, France

    Pierre-Louis Tharaux

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  1. Didier Casanova
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Contributions

D.C., C.B. and T.-L.N. contributed equally. D.C., C.B., P.-L.T. and A.A. conceived and designed the experiments. D.C., C.B. and T.-L.N. performed the experiments. D.C., C.B., T.-L.N. and A.A. analysed the data. R.O.R. and L.B.-S. contributed the cell cultures and data on nanoparticle toxicity. T.G. and J.-P.B. contributed the nanoparticles. All authors discussed the results. C.B. and A.A. co-wrote the paper.

Corresponding author

Correspondence to Antigoni Alexandrou.

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Casanova, D., Bouzigues, C., Nguyên, TL. et al. Single europium-doped nanoparticles measure temporal pattern of reactive oxygen species production inside cells. Nature Nanotech 4, 581–585 (2009). https://doi.org/10.1038/nnano.2009.200

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