Single shot, three-dimensional fluorescence microscopy with a spatially rotating point spread function

Zhaojun Wang, Yanan Cai, Yansheng Liang, Xing Zhou, Shaohui Yan, Dan Dan, Piero R. Bianco, Ming Lei, Baoli Yao

Research output: Contribution to journalArticlepeer-review

12 Scopus citations


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.

Original languageEnglish (US)
Article number#307480
Pages (from-to)5493-5506
Number of pages14
JournalBiomedical Optics Express
Issue number12
StatePublished - Dec 1 2017
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Atomic and Molecular Physics, and Optics


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