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Continuous-wave near-infrared stimulated-emission depletion microscopy using downshifting lanthanide nanoparticles
Nature Nanotechnology  (IF39.213),  Pub Date : 2021-06-14, DOI: 10.1038/s41565-021-00927-y
Liangliang Liang, Ziwei Feng, Qiming Zhang, Thang Do Cong, Yu Wang, Xian Qin, Zhigao Yi, Melgious Jin Yan Ang, Lei Zhou, Han Feng, Bengang Xing, Min Gu, Xiangping Li, Xiaogang Liu

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.