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arXiv · 2403.13436

Multi-photon super-linear image scanning microscopy using upconversion nanoparticles

Yao Wang·Baolei Liu·Lei Ding·Chaohao Chen·Xuchen Shan·Dajing Wang·Menghan Tian·Jiaqi Song·Ze Zheng·Xiaoxue Xu·Xiaolan Zhong·Fan Wang
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Abstract

Super-resolution fluorescence microscopy is of great interest in life science studies for visualizing subcellular structures at the nanometer scale. Among various kinds of super-resolution approaches, image scanning microscopy (ISM) offers a doubled resolution enhancement in a simple and straightforward manner, based on the commonly used confocal microscopes. ISM is also suitable to be integrated with multi-photon microscopy techniques, such as two-photon excitation and second-harmonic generation imaging, for deep tissue imaging, but it remains the twofold limited resolution enhancement and requires expensive femtosecond lasers. Here, we present and experimentally demonstrate the super-linear ISM (SL-ISM) to push the resolution enhancement beyond the factor of two, with a single low-power, continuous-wave, and near-infrared laser, by harnessing the emission nonlinearity within the multiphoton excitation process of lanthanide-doped upconversion nanoparticles (UCNPs). Based on a modified confocal microscope, we achieve a resolution of about 120 nm, 1/8th of the excitation wavelength. Furthermore, we demonstrate a parallel detection strategy of SL-ISM with the multifocal structured excitation pattern, to speed up the acquisition frame rate. This method suggests a new perspective for super-resolution imaging or sensing, multi-photon imaging, and deep-tissue imaging with simple, low-cost, and straightforward implementations.

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BibTeXRIS

Yao Wang, Baolei Liu, Lei Ding, Chaohao Chen, Xuchen Shan, Dajing Wang, Menghan Tian, Jiaqi Song, Ze Zheng, Xiaoxue Xu, Xiaolan Zhong, Fan Wang. 2024-03-20. Multi-photon super-linear image scanning microscopy using upconversion nanoparticles. https://doi.org/10.1002/lpor.202400746

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