Fast multi-directional DSLM for confocal detection without striping artifacts
Authors: Ricci P., Santacaldo G., Gavryusev V., Franceschini A., Mullenbroich MC., Silvestri L., Pavone FS.
Autors Affiliation: European Lab Nonlinear Spect, I-50019 Sesto Fiorentino, Italy; Univ Florence, Dept Phys & Astron, I-50019 Sesto Fiorentino, Italy; Univ Palermo, Dept Phys & Chem, I-90128 Palermo, Italy; Natl Inst Opt, I-50019 Sesto Fiorentino, Italy; Univ Glasgow, Sch Phys & Astron, Glasgow G12 8QQ, Lanark, Scotland
Abstract: In recent years light-sheet fluorescence microscopy (LSFM) has become a cornerstone technology for neuroscience, improving the quality and capabilities of 3D imaging. By selectively illuminating a single plane, it provides intrinsic optical sectioning and fast image recording, while minimizing out of focus fluorescence background, sample photo-damage and photo-bleaching. However, images acquired with LSFM are often affected by light absorption or scattering effects, leading to un-even illumination and striping artifacts. In this work we present an optical solution to this problem, via fast multi-directional illumination of the sample, based on an acousto-optical deflector (AOD). We demonstrate that this pivoting system is compatible with confocal detection in digital scanned laser light-sheet fluorescence microscopy (DSLM) by using a pivoted elliptical-Gaussian beam. We tested its performance by acquiring signals emitted by specific fluorophores in several mouse brain areas, comparing the pivoting beam illumination and a traditional static one, measuring the point spread function response and quantifying the striping reduction. We observed real-time shadow suppression, while preserving the advantages of confocal detection for image contrast. (C) 2020 Optical Society of America under the terms of the OSA Open Access Publishing Agreement
Journal/Review: BIOMEDICAL OPTICS EXPRESS
Volume: 11 (6) Pages from: 3111 to: 3124
KeyWords: LIGHT-SHEET MICROSCOPY; FLUORESCENCE MICROSCOPY; BRAIN ACTIVITY; BESSEL BEAMS; RECONSTRUCTION; SCALEDOI: 10.1364/BOE.390916Citations: 3data from “WEB OF SCIENCE” (of Thomson Reuters) are update at: 2021-02-28References taken from IsiWeb of Knowledge: (subscribers only)Connecting to view paper tab on IsiWeb: Click hereConnecting to view citations from IsiWeb: Click here