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High-resolution three-photon biomedical imaging using doped ZnS nanocrystals

Nature Materials volume 12, pages 359–366 (2013)Cite this article

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Abstract

Three-photon excitation is a process that occurs when three photons are simultaneously absorbed within a luminophore for photo-excitation through virtual states. Although the imaging application of this process was proposed decades ago, three-photon biomedical imaging has not been realized yet owing to its intrinsic low quantum efficiency. We herein report on high-resolution in vitro and in vivo imaging by combining three-photon excitation of ZnS nanocrystals and visible emission from Mn2+ dopants. The large three-photon cross-section of the nanocrystals enabled targeted cellular imaging under high spatial resolution, approaching the theoretical limit of three-photon excitation. Owing to the enhanced Stokes shift achieved through nanocrystal doping, the three-photon process was successfully applied to high-resolution in vivo tumour-targeted imaging. Furthermore, the biocompatibility of ZnS nanocrystals offers great potential for clinical applications of three-photon imaging.

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Figure 1: Synthesis and characterization of ZnS:Mn NCs.
Figure 2: Three-photon luminescence characteristics of ZnS:Mn NCs.
Figure 3: In vitro three-photon imaging of MDA-MB-435 cells targeted by ZnS:Mn NC–Lyp-1-conjugates.
Figure 4: In vivo three-photon imaging of ZnS:Mn NC–RGD conjugates targeted to tumour.
Figure 5: In vivo toxicity evaluation of ZnS:Mn NCs.

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Acknowledgements

We thank A. Lutich (Ludwig-Maximilians-Universtität München) for a preliminary study on multi-photon spectroscopy and W. Ji (The National University of Singapore) for helpful discussions on the three-photon excitation mechanism. We thank M-S. Won at the Korea Basic Science Institute (KBSI) for the electron paramagnetic resonance characterization, and S. Her at KBSI for the review and suggestion of animal experiment design. We acknowledge financial support by the Research Center Program of Institute for Basic Science (IBS) in Korea.

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Author notes

  1. Zdeněk Petrášek & Petra Schwille

    Present address: Present address: Department of Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, Martinsried 82152, Germany,

  2. Jung Ho Yu and Seung-Hae Kwon: These authors contributed equally to this work

Authors and Affiliations

  1. Center for Nanoparticle Research, Institute for Basic Science (IBS), and School of Chemical and Biological Engineering, Seoul National University, Seoul 151-742, Korea

    Jung Ho Yu, Samuel Woojoo Jun, Kwangsoo Shin, Moonkee Choi, Yong Il Park, Hyon Bin Na, Nohyun Lee, Dong Won Lee, Jeong Hyun Kim & Taeghwan Hyeon

  2. Division of Bio-imaging, Korea Basic Science Institute, Chun-Cheon 200-701, Korea

    Seung-Hae Kwon, Ok Kyu Park & Kyeongsoon Park

  3. Biotechnologisches Zentrum der TU Dresden, Dresden 01307, Germany

    Zdeněk Petrášek & Petra Schwille

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Contributions

J.H.Y., S-H.K. and T.H. designed and carried out the experiments, analysed the data and wrote the manuscript. Z.P. and P.S. carried out the FCS study and interpreted the data. S-H.K., O.K.P., J.H.Y. and J.H.K. designed and carried out the multiphoton imaging experiments. J.H.Y., S.W.J., K.S. and D.W.L. carried out the synthesis of the materials. J.H.Y., K.S., M.C. and Y.I.P. carried out the bioconjugation of the NCs. S-H.K., O.K.P. and K.P. carried out the animal experiments. S-H.K., O.K.P., H.B.N. and N.L. carried out the in vitro toxicity evaluation. All authors have reviewed, discussed and approved the results and conclusions of this Article.

Corresponding authors

Correspondence to Jung Ho Yu, Seung-Hae Kwon, Zdeněk Petrášek, Petra Schwille or Taeghwan Hyeon.

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Yu, J., Kwon, SH., Petrášek, Z. et al. High-resolution three-photon biomedical imaging using doped ZnS nanocrystals. Nature Mater 12, 359–366 (2013). https://doi.org/10.1038/nmat3565

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