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Cell. 2018 Feb 22;172(5):1108-1121.e15. doi: 10.1016/j.cell.2018.02.007.

Super-Resolution Imaging of the Extracellular Space in Living Brain Tissue.

Author information

1
University of Bordeaux, 33077 Bordeaux, France; Interdisciplinary Institute for Neuroscience, CNRS UMR 5297, 33077 Bordeaux, France; Department of Neuroscience, University of the Basque Country UPV/EHU, 48940 Leioa, Spain; Achucarro Basque Center for Neuroscience, 48940 Leioa, Spain.
2
University of Bordeaux, 33077 Bordeaux, France; Interdisciplinary Institute for Neuroscience, CNRS UMR 5297, 33077 Bordeaux, France.
3
University of Bordeaux, 33077 Bordeaux, France; Interdisciplinary Institute for Neuroscience, CNRS UMR 5297, 33077 Bordeaux, France. Electronic address: valentin.nagerl@u-bordeaux.fr.

Abstract

The extracellular space (ECS) of the brain has an extremely complex spatial organization, which has defied conventional light microscopy. Consequently, despite a marked interest in the physiological roles of brain ECS, its structure and dynamics remain largely inaccessible for experimenters. We combined 3D-STED microscopy and fluorescent labeling of the extracellular fluid to develop super-resolution shadow imaging (SUSHI) of brain ECS in living organotypic brain slices. SUSHI enables quantitative analysis of ECS structure and reveals dynamics on multiple scales in response to a variety of physiological stimuli. Because SUSHI produces sharp negative images of all cellular structures, it also enables unbiased imaging of unlabeled brain cells with respect to their anatomical context. Moreover, the extracellular labeling strategy greatly alleviates problems of photobleaching and phototoxicity associated with traditional imaging approaches. As a straightforward variant of STED microscopy, SUSHI provides unprecedented access to the structure and dynamics of live brain ECS and neuropil.

KEYWORDS:

STED microscopy; anatomical reconstructions; connectomics; extracellular labelling; extracellular space; interstitial fluid; shadow imaging; structural plasticity; super-resolution imaging; synaptic morphology

PMID:
29474910
DOI:
10.1016/j.cell.2018.02.007
[Indexed for MEDLINE]

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