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

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 straightfor...

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Bibliographic Details
Published inarXiv.org
Main Authors Wang, Yao, Liu, Baolei, Ding, Lei, Chen, Chaohao, Shan, Xuchen, Wang, Dajing, Tian, Menghan, Song, Jiaqi, Zheng, Ze, Xu, Xiaoxue, Zhong, Xiaolan, Wang, Fan
Format Paper
LanguageEnglish
Published Ithaca Cornell University Library, arXiv.org 20.03.2024
Subjects
Continuous radiation
Excitation
Image enhancement
Image resolution
Infrared lasers
Microscopy
Nanoparticles
Photons
Scanning microscopy
Second harmonic generation
Upconversion
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Summary: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.
ISSN:2331-8422