Open AccessHybrid multifocal structured illumination microscopy with enhanced lateral resolution and axial localization capability
Author(s) -
Zhaojun Wang,
Yanan Cai,
Jia Qian,
Tianyu Zhao,
Yansheng Liang,
Dan Dan,
Ming Lei,
Baoli Yao
Publication year - 2020
Publication title -
biomedical optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.391024
Subject(s) - microscopy , optics , materials science , super resolution microscopy , fluorescence microscope , resolution (logic) , microscope , light sheet fluorescence microscopy , optical sectioning , image resolution , point spread function , optical microscope , computer science , fluorescence , artificial intelligence , scanning confocal electron microscopy , physics , scanning electron microscope
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.