Light sheet fluorescence microscopy using axi-symmetric binary phase filters

SUHO RYU, BAEKCHEON SEONG, CHAN WOOL LEE, MIN YONG AHN, WOO TAEK KIM, KWANG MIN CHOE, CHULMIN JOO

Research output: Contribution to journalArticle

Abstract

Light sheet fluorescence microscopy (LSFM) has become an indispensable tool in biomedical studies owing to its depth-sectioning capability and low photo-bleaching. The axial resolution in LSFM is determined mainly by the thickness of the illumination sheet, and a high numerical-aperture lens is thus preferred in the illumination to increase the axial resolution. However, a rapid divergence of the illumination beam limits the effective field-of-view (FoV), that provides high-resolution images. Several strategies have been demonstrated for FoV enhancement, which involve the use of Bessel or Airy beams, for example. However, the generation of these beams requires complicated optical setup or phase filters with continuous phase distributions, which are difficult to manufacture. In contrast, a binary phase filter (BPF) comprising concentric rings with 0 or π phases produces a response similar to its continuous original and is easy to realize. Here, we present a novel form of LSFM that integrates BPFs derived from two representative axi-symmetric aberrations, including phase axicon and spherical aberrations, to improve the imaging performance. We demonstrate that these BPFs significantly increase the FoV, and those derived from axicon generate self-reconstructing beams, which are highly desirable in imaging through scattering specimens. We validate its high-contrast imaging capability over extended FoV by presenting three-dimensional images of microspheres, imaginal disc of Drosophila larva, and Arabidopsis.

Original languageEnglish
Pages (from-to)3936-3951
Number of pages16
JournalBiomedical Optics Express
Volume11
Issue number7
DOIs
Publication statusPublished - 2020 Jul 1

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Atomic and Molecular Physics, and Optics

Fingerprint Dive into the research topics of 'Light sheet fluorescence microscopy using axi-symmetric binary phase filters'. Together they form a unique fingerprint.

  • Cite this