title: Selectable light-sheet uniformity using tuned axial scanning.
abstract: Light-sheet fluorescence microscopy (LSFM) is an optical sectioning technique capable of rapid three-dimensional (3D) imaging of a wide range of specimens with reduced phototoxicity and superior background rejection. However, traditional light-sheet generation approaches based on elliptical or circular Gaussian beams suffer an inherent trade-off between light-sheet thickness and area over which this thickness is preserved. Recently, an increase in light-sheet uniformity was demonstrated using rapid biaxial Gaussian beam scanning along the lateral and beam propagation directions. Here we apply a similar scanning concept to an elliptical beam generated by a cylindrical lens. In this case, only z-scanning of the elliptical beam is required and hence experimental implementation of the setup can be simplified. We introduce a simple dimensionless uniformity statistic to better characterize scanned light-sheets and experimentally demonstrate custom tailored uniformities up to a factor of 5 higher than those of unscanned elliptical beams. This technique offers a straightforward way to generate and characterize a custom illumination profile that provides enhanced utilization of the detector dynamic range and field of view, opening the door to faster and more efficient 2D and 3D imaging.
authors.affiliation: Department of Nanophysics, Istituto Italiano di Tecnologia, Genoa, Via Morego 30, 16163, Italy.
authors.forename: Martí
authors.initials: M
authors.lastname: Duocastella
authors.affiliation: Department of Mechanical and Aerospace Engineering, Princeton University, Olden St, Princeton, NJ, 08544, USA.
authors.forename: Craig B
authors.initials: CB
authors.lastname: Arnold
authors.affiliation: Department of Physics, Princeton University, Washington Avenue, Princeton, NJ, 08544, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Puchalla
keywords: (180.2520) fluorescence microscopy; (180.6900) three-dimensional microscopy; OCIS codes: (110.1080) active or adaptive optics