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bioRxiv · 10.1101/2024.09.02.610873

Scattering-Based Super-Resolution Optical Fluctuation Imaging

Yudovich, S.·Posnjak, G.·Shani, L.·Teblum, E.·Liedl, T.·Enderlein, J.·Weiss, S.
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Abstract

Super-resolution optical imaging has become a prominent tool in life and material sciences, allowing one to decipher structures at increasingly greater spatial detail. Among the utilized techniques in this field, super-resolution optical fluctuation imaging (SOFI) has proved to be a valuable approach. A major advantage of SOFI is its less restrictive requirements for generating super-resolved images of neighboring nanostructures or molecules, as it only assumes that the detected fluctuating light from neighboring emitters is statistically uncorrelated, but not necessarily separated in time. While most optical super-resolution microscopies depend on signals obtained from fluorescence, they are limited by photobleaching and phototoxicity. An alternative source for optical signals can be acquired by detecting the light scattered from molecules or nanoparticles. However, the application of coherent scattering-based imaging modalities for super-resolution imaging has been considerably limited compared to fluorescence-based modalities. Here, we develop scattering-based super-resolution optical fluctuation imaging (sSOFI), where we utilize the rotation of anisotropic particles as a source of fluctuating optical signals. We discuss the differences in the application of SOFI algorithms for coherent and incoherent imaging modalities, and utilize interference microscopy to demonstrate super-resolution imaging of rotating nanoparticle dimers. We present a theoretical analysis of the relevant model systems, and discuss the possible effects of cusp artifacts and electrodynamic coupling between nearby nano-scatterers. Finally, we apply sSOFI as a label-free novelty filter that highlights regions with higher activity of biomolecules and demonstrate its use by imaging membrane protrusions of live cells. Overall, the development of optical super-resolution approaches for coherent scattering-based imaging modalities, as described here, could potentially allow for the investigation of biological processes at temporal resolutions and acquisition durations previously inaccessible in fluorescence-based imaging.

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Yudovich, S., Posnjak, G., Shani, L., Teblum, E., Liedl, T., Enderlein, J., Weiss, S.. 2024-09-03. Scattering-Based Super-Resolution Optical Fluctuation Imaging. https://doi.org/10.1101/2024.09.02.610873

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