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

Publication date: 22 February 2018 Source:Cell, Volume 172, Issue 5 Author(s): Jan Tønnesen, V.V.G. Krishna Inavalli, U. Valentin Nägerl 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. Graphical abstract Teaser Live tissue super-resolution 3D-STED microscopy combined with fluorescence labeling of the interstitial fluid reveals the complex spatial organization of the extracellular space of the brain.

Source: Cell - February 24, 2018 Category: Cytology Source Type: research