Fast multi-directional DSLM for confocal detection without striping artifacts

Giuseppe Sancataldo, Francesco Saverio Pavone, Marie Caroline Müllenbroich, Ludovico Silvestri, Giuseppe Sancataldo, Vladislav Gavryusev, Alessandra Franceschini, Pietro Ricci

Risultato della ricerca: Articlepeer review

3 Citazioni (Scopus)

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.
Lingua originaleEnglish
pagine (da-a)3111-3124
Numero di pagine14
RivistaBiomedical Optics Express
Volume11
Stato di pubblicazionePublished - 2020

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Atomic and Molecular Physics, and Optics

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