Frontiers in Neuroscience (Oct 2022)

Axial resolution and imaging contrast enhancement in inverted light-sheet microscopy by natural illumination modulation

  • Zhi Wang,
  • Zhi Wang,
  • Wei Qiao,
  • Wei Qiao,
  • Tao Jiang,
  • Siqi Chen,
  • Siqi Chen,
  • Bolin Lu,
  • Bolin Lu,
  • Kefu Ning,
  • Kefu Ning,
  • Rui Jin,
  • Rui Jin,
  • Hui Gong,
  • Hui Gong,
  • Hui Gong,
  • Jing Yuan,
  • Jing Yuan,
  • Jing Yuan

DOI
https://doi.org/10.3389/fnins.2022.1032195
Journal volume & issue
Vol. 16

Abstract

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Inverted light-sheet microscopy (ILSM) is widely employed for fast large-volume imaging of biological tissue. However, the scattering especially in an uncleared sample, and the divergent propagation of the illumination beam lead to a trade-off between axial resolution and imaging depth. Herein, we propose naturally modulated ILSM (NM-ILSM) as a technique to improve axial resolution while simultaneously maintaining the wide field-of-view (FOV), and enhancing imaging contrast via background suppression. Theoretical derivations, simulations, and experimental imaging demonstrate 15% axial resolution increases, and fivefold greater image contrast compared with conventional ILSM. Therefore, NM-ILSM allows convenient imaging quality improvement for uncleared tissue and could extend the biological application scope of ILSM.

Keywords