Adaptive optics in single objective inclined light sheet microscopy enables three-dimensional localization microscopy in adult Drosophila brains

S. Hung, Arnau Llobet Rosell, Daphne Jurriens, O.A. Soloviev, Lukas C. Kapitein, K.S. Grußmayer, Lukas J. Neukomm, M.H.G. Verhaegen, C.S. Smith*

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

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Abstract

Single-molecule localization microscopy (SMLM) enables the high-resolution visualization of organelle structures and the precise localization of individual proteins. However, the expected resolution is not achieved in tissue as the imaging conditions deteriorate. Sample-induced aberrations distort the point spread function (PSF), and high background fluorescence decreases the localization precision. Here, we synergistically combine sensorless adaptive optics (AO), in-situ 3D-PSF calibration, and a single-objective lens inclined light sheet microscope (SOLEIL), termed (AO-SOLEIL), to mitigate deep tissue-induced deteriorations. We apply AO-SOLEIL on several dSTORM samples including brains of adult Drosophila. We observed a 2x improvement in the estimated axial localization precision with respect to widefield without aberration correction while we used synergistic solution. AO-SOLEIL enhances the overall imaging resolution and further facilitates the visualization of sub-cellular structures in tissue.
Original languageEnglish
Article number954949
Number of pages21
JournalFrontiers in Neuroscience
Volume16
DOIs
Publication statusPublished - 2022

Keywords

  • Super-resolution Microscopy
  • localization microscopy
  • adaptive optics
  • Drosophila
  • brain

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