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Continuous-wave near-infrared stimulated-emission depletion microscopy using downshifting lanthanide nanoparticles

Nature Nanotechnology volume 16, pages 975–980 (2021)Cite this article

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Abstract

Stimulated-emission depletion (STED) microscopy has profoundly extended our horizons to the subcellular level1,2,3. However, it remains challenging to perform hours-long, autofluorescence-free super-resolution imaging in near-infrared (NIR) optical windows under facile continuous-wave laser depletion at low power4,5. Here we report downshifting lanthanide nanoparticles that enable background-suppressed STED imaging in all-NIR spectral bands (λexcitation = 808 nm, λdepletion = 1,064 nm and λemission = 850–900 nm), with a lateral resolution of below 20 nm and zero photobleaching. With a quasi-four-level configuration and long-lived (τ > 100 μs) metastable states, these nanoparticles support near-unity (98.8%) luminescence suppression under 19 kW cm−2 saturation intensity. The all-NIR regime enables high-contrast deep-tissue (~50 μm) imaging with approximately 70 nm spatial resolution. These lanthanide nanoprobes promise to expand the application realm of STED microscopy and pave the way towards high-resolution time-lapse investigations of cellular processes at superior spatial and temporal dimensions.

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Fig. 1: All-NIR quasi-four-level CW STED microscopy.
Fig. 2: Optical switching features of neodymium-activated NIR STED nanoprobes.
Fig. 3: Mechanistic investigation of the neodymium-activated nanocrystals for superior STED.
Fig. 4: Low-power full-NIR CW STED imaging of subcellular structures and deep-tissue super-resolution imaging.

Data availability

Source data are provided with this paper. The data that support the findings of this study are available within the article and its Supplementary Information. Additional data are available from the corresponding authors upon request.

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Acknowledgements

This work is supported by the National Key R&D Program of China (no. 2018YFA0902600 and no. 2018YFB1107200), the National Natural Science Foundation of China (21635002, 21771135, 21871071 and 61975123), the Ministry of Education, Singapore (MOE2017-T2-2-110), the Agency for Science, Technology and Research (A*STAR) (grant no. A1883c0011 and no. A1983c0038), National Research Foundation, the Prime Minister’s Office of Singapore under its NRF Investigatorship Programme (award no. NRF-NRFI05-2019-0003), the National Basic Research Program of China (973 Program, grant no. 2015CB932200), Zhangjiang National Innovation Demonstration Zone (ZJ-2019-ZD-005) and the Guangdong Provincial Innovation and Entrepreneurship Project (grant no. 2016ZT06D081). We thank X. Zhao, J. Chen and Z. Mu for technical assistance with the TEM imaging and NIR spectral measurements. We also thank J. Hong for absorption spectra measurements.

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Author notes

  1. These authors contributed equally: Liangliang Liang, Ziwei Feng, Qiming Zhang.

Authors and Affiliations

  1. Department of Chemistry, National University of Singapore, Singapore, Singapore

    Liangliang Liang, Xian Qin, Zhigao Yi, Melgious Jin Yan Ang, Lei Zhou & Xiaogang Liu

  2. Guangdong Provincial Key Laboratory of Optical Fiber Sensing and Communications, Institute of Photonics Technology, Jinan University, Guangzhou, China

    Ziwei Feng & Xiangping Li

  3. Centre for Artificial-Intelligence Nanophotonics, School of Optical-Electrical and Computer Engineering, University of Shanghai for Science and Technology, Shanghai, China

    Qiming Zhang & Min Gu

  4. Division of Chemistry and Biological Chemistry, School of Physical & Mathematical Sciences, Nanyang Technological University, Singapore, Singapore

    Thang Do Cong & Bengang Xing

  5. SZU-NUS Collaborative Innovation Center for Optoelectronic Science & Technology, Institute of Microscale Optoelectronics, Shenzhen University, Shenzhen, China

    Yu Wang & Xiaogang Liu

  6. Nanyang Environment and Water Research Institute, Interdisciplinary Graduate Programme, Nanyang Technological University, Singapore, Singapore

    Han Feng

  7. Joint School of National University of Singapore and Tianjin University, International Campus of Tianjin University, Fuzhou, China

    Xiaogang Liu

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Contributions

L.L. and X. Liu conceived and designed the project. X. Liu, X. Li, M.G. and B.X. supervised the project and led the collaboration efforts. L.L. synthesized the nanocrystals and conducted the numerical simulations with contribution from L.Z., Q.Z. and Z.F. Optical experiments and super-resolution imaging were conducted by L.L., Z.F. and Y.W. The preparation of mouse-brain slices and cell labelling was the responsibility of Z.Y., M.J.Y.A., T.D.C. and H.F. The density functional theory calculations were conducted by X.Q. The manuscript was written by L.L., Z.F. and X. Liu. All authors participated in the discussion and analysis of the manuscript.

Corresponding authors

Correspondence to Min Gu, Xiangping Li or Xiaogang Liu.

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Liang, L., Feng, Z., Zhang, Q. et al. Continuous-wave near-infrared stimulated-emission depletion microscopy using downshifting lanthanide nanoparticles. Nat. Nanotechnol. 16, 975–980 (2021). https://doi.org/10.1038/s41565-021-00927-y

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