Open AccessSingle shot, three-dimensional fluorescence microscopy with a spatially rotating point spread function
Author(s) -
Zhaojun Wang,
Yanan Cai,
Yansheng Liang,
Xing Zhou,
Shaohui Yan,
Dan Dan,
Piero R. Bianco,
Ming Lei,
Baoli Yao
Publication year - 2017
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.8.005493
Subject(s) - optics , point spread function , microscope , light sheet fluorescence microscopy , microscopy , depth of field , optical microscope , materials science , optical sectioning , numerical aperture , fluorescence microscope , holography , physics , fluorescence , scanning electron microscope , scanning confocal electron microscopy , wavelength
A wide-field fluorescence microscope with a double-helix point spread function (PSF) is constructed to obtain the specimen's three-dimensional distribution with a single snapshot. Spiral-phase-based computer-generated holograms (CGHs) are adopted to make the depth-of-field of the microscope adjustable. The impact of system aberrations on the double-helix PSF at high numerical aperture is analyzed to reveal the necessity of the aberration correction. A modified cepstrum-based reconstruction scheme is promoted in accordance with properties of the new double-helix PSF. The extended depth-of-field images and the corresponding depth maps for both a simulated sample and a tilted section slice of bovine pulmonary artery endothelial (BPAE) cells are recovered, respectively, verifying that the depth-of-field is properly extended and the depth of the specimen can be estimated at a precision of 23.4nm. This three-dimensional fluorescence microscope with a framerate-rank time resolution is suitable for studying the fast developing process of thin and sparsely distributed micron-scale cells in extended depth-of-field.