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Super-resolution imaging of the extracellular space in living brain tissue - March 2018

Super-resolution imaging of the extracellular space in living brain tissue - March 2018

JanTønnesen, V.V.G. KrishnaInavalli, Valentin Nägerl
Cell 2018 Feb 22;172(5):1108-1121.e15. doi: 10.1016/j.cell.2018.02.007 - Free access here

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.

Fig 1: Super-resolution shadow imaging (SUSHI) renders all brain cells visible ‘in one shot’. The image on the left shows an overview of neurons in the hippocampus, which is the archetypical memory center of the mammalian brain. At higher zoom, the image on the right reveals the ‘cable tangle’ of brain tissue with nanoscale spatial resolution. The yellow neuron was labeled with a fluorescent protein and thus stands out from the rest of the inversely labeled tissue.

Contact: Valentin Nagerl

See more details in the French CNRS press release and on Bordeaux Neurocampus website