Light-sheet microscopy by confocal line scanning of dual-Bessel beams

Zhang, Pengfei; Phipps, Mary Elizabeth; Goodwin, Peter Marvin; Werner, James H.

doi:10.1117/1.JBO.21.10.100502

Light-sheet microscopy by confocal line scanning of dual-Bessel beams

Journal Article · Tue Oct 25 00:00:00 EDT 2016 · Journal of Biomedical Optics

DOI:https://doi.org/10.1117/1.JBO.21.10.100502· OSTI ID:1356125

Zhang, Pengfei ^[1]; Phipps, Mary Elizabeth ^[2]; ^[2]; ^[2]

Washington Univ., St. Louis, MO (United States)
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

Here, we have developed a light-sheet microscope that uses confocal scanning of dual-Bessel beams for illumination. A digital micromirror device (DMD) is placed in the intermediate image plane of the objective used to collect fluorescence and is programmed with two lines of pixels in the “on” state such that the DMD functions as a spatial filter to reject the out-of-focus background generated by the side-lobes of the Bessel beams. The optical sectioning and out-of-focus background rejection capabilities of this microscope were demonstrated by imaging of fluorescently stained actin in human A431 cells. The dual-Bessel beam system enables twice as many photons to be detected per imaging scan, which is useful for low light applications (e.g., single-molecule localization) or imaging at high speed with a superior signal to noise. While demonstrated for two Bessel beams, this approach is scalable to a larger number of beams.

View Accepted Manuscript (DOE)

Research Organization:: Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

Sponsoring Organization:: LDRD; USDOE

Grant/Contract Number:: AC52-06NA25396

OSTI ID:: 1356125

Report Number(s):: LA-UR--15-24283

Journal Information:: Journal of Biomedical Optics, Journal Name: Journal of Biomedical Optics Journal Issue: 10 Vol. 21; ISSN 1083-3688

Publisher:: SPIECopyright Statement

Country of Publication:: United States

Language:: English

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Cited By (3)

Technical implementations of light sheet microscopy Elisa, Zagato; Toon, Brans; De Smedt, Stefaan C. Microscopy Research and Technique, Vol. 81, Issue 9 https://doi.org/10.1002/jemt.22981	journal	January 2018
Minutes-timescale 3D isotropic imaging of entire organs at subcellular resolution by content-aware compressed-sensing light-sheet microscopy Fang, Chunyu; Chu, Tingting; Yu, Tingting Nature Communications https://doi.org/10.1101/825901	journal	April 2020
Light sheet approaches for improved precision in 3D localization-based super-resolution imaging in mammalian cells [Invited] Gustavsson, Anna-Karin; Petrov, Petar N.; Moerner, W. E. Optics Express, Vol. 26, Issue 10 https://doi.org/10.1364/oe.26.013122	journal	January 2018

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