Open AccessFast multi-directional DSLM for confocal detection without striping artifacts
Author(s) -
Pietro Ricci,
Giuseppe Sancataldo,
Vladislav Gavryusev,
Alessandra Franceschini,
Marie Caroline Müllenbroich,
Ludovico Silvestri,
Francesco S. Pavone
Publication year - 2020
Publication title -
biomedical optics express
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.390916
Subject(s) - light sheet fluorescence microscopy , optics , optical sectioning , confocal , digital micromirror device , point spread function , confocal microscopy , data striping , materials science , microscopy , light scattering , image quality , cardinal point , microscope , computer science , scattering , artificial intelligence , physics , scanning confocal electron microscopy , image (mathematics) , operating system
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.