Fast multi-directional DSLM for confocal detection without striping artifacts

Ricci, P., Sancataldo, G., Gavryusev, V., Franceschini, A., Müllenbroich, M. C. , Silvestri, L. and Pavone, F. S. (2020) Fast multi-directional DSLM for confocal detection without striping artifacts. Biomedical Optics Express, 11(6), pp. 3111-3124. (doi: 10.1364/BOE.390916) (PMID:32637245) (PMCID:PMC7316030)

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.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Muellenbroich, Dr Caroline
• Authors: Ricci, P., Sancataldo, G., Gavryusev, V., Franceschini, A., Müllenbroich, M. C., Silvestri, L., and Pavone, F. S.
• College/School: College of Science and Engineering > School of Physics and Astronomy
• Journal Name: Biomedical Optics Express
• ISSN: 2156-7085
• Published Online: 18 May 2020
• Copyright Holders: Copyright © 2020 Optical Society of America
• First Published: First published in Biomedical Optics Express 11(6): 3111-3124
• Publisher Policy: Reproduced in accordance with the publisher copyright policy