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Lattice light-sheet microscopy and photo-stimulation in brain slices

Abstract : Lattice light sheet (LLS) fluorescence microscopy is a powerful recent technique for in vivo imaging of single and multi-cellular samples at very high spatio-temporal resolutions. We built a LLS microscope in which we added a photo-stimulation 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.
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https://www.hal.inserm.fr/inserm-03016227
Contributor : Mathieu Ducros <>
Submitted on : Friday, November 20, 2020 - 12:09:37 PM
Last modification on : Tuesday, November 24, 2020 - 10:00:06 PM

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Mathieu Ducros, Angela Getz, Misa Arizono, Valeria Pecoraro, Monica Fernandez Monreal, et al.. Lattice light-sheet microscopy and photo-stimulation in brain slices. Neural Imaging and Sensing 2019, Feb 2019, San Francisco, France. pp.8, ⟨10.1117/12.2509467⟩. ⟨inserm-03016227⟩

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