Hybrid multifocal structured illumination microscopy with enhanced lateral resolution and axial localization capability

PLUTO / PLUTO-2 Spatial Light Modulators
Microscopy Misc.: Speckle / Characterization / Metrology
Published on:
Authors: Wang, Zhaojun; Cai, Yanan; Qian, Jia; Zhao, Tianyu; Liang, Yansheng; Dan, Dan; Lei, Ming & Yao, Baoli
Abstract:

“Super-resolution (SR) fluorescence microscopy that breaks through the diffraction barrier has drawn great interest in biomedical research. However, obtaining a high precision three-dimensional distribution of the specimen in a short time still remains a challenging task for existing techniques. In this paper, we propose a super-resolution fluorescence microscopy with axial localization capability by combining multifocal structured illumination microscopy with a hybrid detection PSF composed of a Gaussian PSF and a double-helix PSF. A modified reconstruction scheme is presented to accommodate the new hybrid PSF. This method can not only recover the lateral super-resolution image of the specimen but also retain the specimen’s depth map within a range of 600 nm with an axial localization precision of 20.8 nm. The performance of this approach is verified by testing fluorescent beads and tubulin in 293-cells. The developed microscope is well suited for observing the precise 3D distribution of thin specimens.”

Open Access

Publication: Biomedical Optics Express
Issue/Year: Biomedical Optics Express, Volume 11; Number 6; Pages 3058; 2020
DOI: 10.1364/boe.391024
Link: https://doi.org/10.1364/boe.391024

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