Fast multi-directional DSLM for confocal detection without striping artifacts

Year: 2020

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

More Information: Horizon 2020 Framework Programme (654148, 720270, 785907); H2020 Excellent Science (692943); Ministero dellŽIstruzione, dellŽUniversita e della Ricerca Flagship Project NanoMax; Eurobioimaging Italian Nodes (ESFRI); Progetto ordinario di Ricerca Finalizzata del Ministero della salute (RF-2013-02355240); Ente Cassa di Risparmio di Firenze; H2020 Marie SklodowskaCurie Actions (793849, 842893).
KeyWords: LIGHT-SHEET MICROSCOPY; FLUORESCENCE MICROSCOPY; BRAIN ACTIVITY; BESSEL BEAMS; RECONSTRUCTION; SCALE
DOI: 10.1364/BOE.390916

Citations: 15
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