Frontiers in Neuroanatomy (Feb 2019)

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

  • Giuseppe Sancataldo,
  • Giuseppe Sancataldo,
  • Vladislav Gavryusev,
  • Vladislav Gavryusev,
  • Giuseppe de Vito,
  • Giuseppe de Vito,
  • Lapo Turrini,
  • Lapo Turrini,
  • Massimiliano Locatelli,
  • Massimiliano Locatelli,
  • Chiara Fornetto,
  • Chiara Fornetto,
  • Natascia Tiso,
  • Francesco Vanzi,
  • Francesco Vanzi,
  • Ludovico Silvestri,
  • Ludovico Silvestri,
  • Francesco Saverio Pavone,
  • Francesco Saverio Pavone

DOI
https://doi.org/10.3389/fnana.2019.00007
Journal volume & issue
Vol. 13

Abstract

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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 zebrafish 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.

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