Achieving Low-Power Single-Wavelength-pair Nanoscopy with NIR-II Continuous-Wave Laser for Multi-Chromatic Probes

Stimulated emission depletion (STED) microscopy is a powerful diffraction-unlimited technique for fluorescence imaging. Despite its rapid evolution, STED fundamentally suffers from high-intensity light illumination, sophisticated probe-defined laser schemes, and limited photon budget of the probes. Here, we demonstrate a versatile strategy, stimulated-emission induced excitation depletion (STExD), to deplete the emission of multi-chromatic probes using a single pair of low-power, near-infrared (NIR), continuous-wave (CW) lasers with fixed wavelengths. With the effect of cascade amplified depletion in lanthanide upconversion systems, we achieve emission inhibition for a wide range of emitters ( e.g ., Nd 3+ , Yb 3+ , Er 3+ , Ho 3+ , Pr 3+ , Eu 3+ , Tm 3+ , Gd 3+ , and Tb 3+ ) by manipulating their common sensitizer, i.e., Nd 3+ ions, using a 1064-nm laser. With NaYF 4 :Nd nanoparticles, we demonstrate an ultrahigh depletion efficiency of 99.3 ± 0.3% for the 450 nm emission with a low saturation intensity of 23.8 ± 0.4 kW cm −2 . We further demonstrate nanoscopic imaging with a series of multi-chromatic nanoprobes with a lateral resolution down to 34 nm, two-color STExD imaging, and subcellular imaging of the immunolabelled actin filaments. The strategy expounded here promotes single wavelength-pair nanoscopy for multi-chromatic probes and for multi-color imaging under low-intensity-level NIR-II CW laser depletion.