Flexible Multi-Beam Light-Sheet Fluorescence Microscope for Live Imaging Without Striping Artifacts

Year: 2019

Authors: Sancataldo G., Gavryusev V., de Vito G., Turrini L., Locatelli M., Fornetto C., Tiso N., Vanzi F., Silvestri L., Pavone FS.

Autors Affiliation: Univ Florence, Dept Phys & Astron, Sesto Fiorentino, Italy; Univ Florence, European Lab Nonlinear Spect, Sesto Fiorentino, Italy; CNR, Natl Inst Opt, Sesto Fiorentino, Italy; Univ Padua, Dept Biol, Padua, Italy; Univ Florence, Dept Biol, Sesto Fiorentino, Italy

Abstract: The development of light-sheet fluorescence microscopy (LSFM) has greatly expanded the experimental capabilities in many biological and biomedical research fields, enabling for example live studies of murine and zebra fish neural activity or of cell growth and division. The key feature of the method is the selective illumination of a sample single plane, providing an intrinsic optical sectioning and allowing direct 2D image recording. On the other hand, this excitation scheme is more affected by absorption or scattering artifacts in comparison to point scanning methods, leading to un-even illumination. We present here an easily implementable method, based on acousto-optical deflectors (AOD), to overcome this obstacle. We report the advantages provided by flexible and fast AODs in generating simultaneous angled multiple beams from a single laser beam and in fast light sheet pivoting and we demonstrate the suppression of illumination artifacts.

Journal/Review: FRONTIERS IN NEUROANATOMY

Volume: 13      Pages from: 7-1  to: 7-8

More Information: This project has received funding from the European Union´s Horizon 2020 Framework Programme for Research and Innovation under grant agreement No. 654148 (Laserlab-Europe), specific grant agreements No. 720270 (Human Brain Project SGA1), No. 785907 (Human Brain Project SGA2), and No. 793849 (MesoBrainMicr), from the H2020 EXCELLENT SCIENCE-European Research Council (ERC) under grant agreement ID No. 692943 (BrainBIT) and from Eurobioimaging-Advanced Light Microscopy Italian Node.
KeyWords: light-sheet fluorescence microscopy; striping artifacts; fast volumetric imaging; acousto optic deflector; brain imaging; zebrafish
DOI: 10.3389/fnana.2019.00007

Citations: 20
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