title: Light-sheet microscopy by confocal line scanning of dual-Bessel beams.
abstract: 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.
authors: Washington University in St. Louis, Department of Biomedical Engineering, Optical Imaging Laboratory, One Brookings Drive, St. Louis, Missouri 63130-4899, United States.
authors: Pengfei
authors: P
authors: Zhang
authors: Center for Integrated Nanotechnologies, Los Alamos National Laboratory, Mail Stop G755, Los Alamos, New Mexico 87545, United States.
authors: Mary E
authors: ME
authors: Phipps
authors: Center for Integrated Nanotechnologies, Los Alamos National Laboratory, Mail Stop G755, Los Alamos, New Mexico 87545, United States.
authors: Peter M
authors: PM
authors: Goodwin
authors: Center for Integrated Nanotechnologies, Los Alamos National Laboratory, Mail Stop G755, Los Alamos, New Mexico 87545, United States.
authors: James H
authors: JH
authors: Werner
mesh_terms: D002460:Cell Line; D004867:Equipment Design; D006651:Histocytochemistry; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D017785:Photons