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Lattice light sheet microscopy and photo-stimulation in brain slices

  • Mathieu Ducros
  • , Angela Getz
  • , Misa Arizono
  • , Valeria Pecoraro
  • , Monica Fernandez-Monreal
  • , Mathieu Letellier
  • , U. Valentin Nägerl
  • , Daniel Choquet

Research output: Chapter in Book / Report / Conference proceedingConference contributionAcademicpeer-review

Abstract

Lattice light sheet (LLS) fluorescence microscopy is a powerful recent technique for in vivo imaging of single and multicellular samples at very high spatiooral resolutions. We built a LLS microscope in which we added a photostimulation path to perform all-optical neurophysiological studies in rodent hippocampal brain slices. Thanks to the photo-stimulation path we could achieve fluorescence recovery after photobleaching (FRAP) or glutamate uncaging at spatially and temporally controlled regions of interest. Several fluorescence labelling protocols were employed depending on the imaged structure. Sub-micrometric neuronal elements such as spines or dendritic vesicles could be imaged down to ∼20 μm below the surface. We demonstrate the performances of LLS in several ongoing studies: measurement of AMPA receptor surface diffusion at single spines, vesicular transport in dendrites, spontaneous and stimulated local calcium activity in neurons and astrocytes.
Original languageEnglish
Title of host publicationNeural Imaging and Sensing 2019
EditorsL. Fu, Q. Luo, J. Ding
PublisherSPIE
ISBN (Electronic)9781510623729
DOIs
Publication statusPublished - 2019
Externally publishedYes
EventNeural Imaging and Sensing 2019 - San Francisco, United States
Duration: 4 Feb 20195 Feb 2019

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
ISSN (Print)1605-7422

Conference

ConferenceNeural Imaging and Sensing 2019
Country/TerritoryUnited States
CitySan Francisco
Period4/02/195/02/19

Funding

We thank the Howard Hughes Medical Institute, Janelia Research Campus for providing all the technical information to reproduce the lattice light sheet microscope upon signature of a research license agreement. Among the HHMI staff, Westley R. Legant and Daniel E Milkie provided very precious help to build the setup and handle the controlling software. We thanks all the Bordeaux Imaging photonic team for technical support. We thank Julie Angibaud for preparing organotypic slices with the rolling tube technique. This work was financially supported by France Bioimaging, the GIS IBiSA and DynSynMem ERC grant.

Funders
DynSynMem ERC

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