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Light-sheet fluorescence expansion microscopy: fast mapping of neural circuits at super resolution.

Jana BürgersIrina PavlovaJuan E Rodriguez-GaticaChristian HennebergerMarc OellerJan A RulandJan P SiebrasseUlrich KubitscheckMartin K Schwarz
Published in: Neurophotonics (2019)
The goal of understanding the architecture of neural circuits at the synapse level with a brain-wide perspective has powered the interest in high-speed and large field-of-view volumetric imaging at subcellular resolution. Here, we developed a method combining tissue expansion and light-sheet fluorescence microscopy to allow extended volumetric super resolution high-speed imaging of large mouse brain samples. We demonstrate the capabilities of this method by performing two color fast volumetric super resolution imaging of mouse CA1 and dentate gyrus molecular-, granule cell-, and polymorphic layers. Our method enables an exact evaluation of granule cell and neurite morphology within the context of large cell ensembles spanning several orders of magnitude in resolution. We found that imaging a brain region of 1    mm 3 in super resolution using light-sheet fluorescence expansion microscopy is about 17-fold faster than imaging the same region by a current state-of-the-art high-resolution confocal laser scanning microscope.
Keyphrases
  • high resolution
  • high speed
  • single molecule
  • atomic force microscopy
  • mass spectrometry
  • single cell
  • cell therapy
  • stem cells
  • optical coherence tomography
  • white matter
  • multiple sclerosis
  • resting state