Apr 29, 2020

Fast multi-directional DSLM for confocaldetection without striping artifacts

BioRxiv : the Preprint Server for Biology
Pietro RicciF. S. Pavone

Abstract

In recent years Light-Sheet Fluorescence Microscopy (LSFM) has become a cornerstone technology for neuroscience, improving quality and capabilities of 3D imaging. By illuminating selectively 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, in terms of Point Spread Function response, striping reduction and contrast improvement. We observed real-time shadow suppression while preserving confocal detection advantages for image cont...Continue Reading

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