Multi-photon near-infrared emission saturation nanoscopy using upconversion nanoparticles

Chaohao Chen, Fan Wang (), Shihui Wen, Qian Peter Su, Mike C. L. Wu, Yongtao Liu, Baoming Wang, Du Li, Xuchen Shan, Mehran Kianinia, Igor Aharonovich, Milos Toth, Shaun P. Jackson, Peng Xi and Dayong Jin ()
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Chaohao Chen: University of Technology Sydney
Fan Wang: University of Technology Sydney
Shihui Wen: University of Technology Sydney
Qian Peter Su: University of Technology Sydney
Mike C. L. Wu: University of Sydney
Yongtao Liu: University of Technology Sydney
Baoming Wang: University of Technology Sydney
Du Li: University of Technology Sydney
Xuchen Shan: University of Technology Sydney
Mehran Kianinia: University of Technology Sydney
Igor Aharonovich: University of Technology Sydney
Milos Toth: University of Technology Sydney
Shaun P. Jackson: University of Sydney
Peng Xi: Peking University
Dayong Jin: University of Technology Sydney

Nature Communications, 2018, vol. 9, issue 1, 1-6

Abstract: Abstract Multiphoton fluorescence microscopy (MPM), using near infrared excitation light, provides increased penetration depth, decreased detection background, and reduced phototoxicity. Using stimulated emission depletion (STED) approach, MPM can bypass the diffraction limitation, but it requires both spatial alignment and temporal synchronization of high power (femtosecond) lasers, which is limited by the inefficiency of the probes. Here, we report that upconversion nanoparticles (UCNPs) can unlock a new mode of near-infrared emission saturation (NIRES) nanoscopy for deep tissue super-resolution imaging with excitation intensity several orders of magnitude lower than that required by conventional MPM dyes. Using a doughnut beam excitation from a 980 nm diode laser and detecting at 800 nm, we achieve a resolution of sub 50 nm, 1/20th of the excitation wavelength, in imaging of single UCNP through 93 μm thick liver tissue. This method offers a simple solution for deep tissue super resolution imaging and single molecule tracking.

Date: 2018
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DOI: 10.1038/s41467-018-05842-w

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