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Publication : Dense 4D nanoscale reconstruction of living brain tissue.

First Author  Velicky P Year  2023
Journal  Nat Methods Volume  20
Issue  8 Pages  1256-1265
PubMed ID  37429995 Mgi Jnum  J:353164
Mgi Id  MGI:7663236 Doi  10.1038/s41592-023-01936-6
Citation  Velicky P, et al. (2023) Dense 4D nanoscale reconstruction of living brain tissue. Nat Methods 20(8):1256-1265
abstractText  Three-dimensional (3D) reconstruction of living brain tissue down to an individual synapse level would create opportunities for decoding the dynamics and structure-function relationships of the brain's complex and dense information processing network; however, this has been hindered by insufficient 3D resolution, inadequate signal-to-noise ratio and prohibitive light burden in optical imaging, whereas electron microscopy is inherently static. Here we solved these challenges by developing an integrated optical/machine-learning technology, LIONESS (live information-optimized nanoscopy enabling saturated segmentation). This leverages optical modifications to stimulated emission depletion microscopy in comprehensively, extracellularly labeled tissue and previous information on sample structure via machine learning to simultaneously achieve isotropic super-resolution, high signal-to-noise ratio and compatibility with living tissue. This allows dense deep-learning-based instance segmentation and 3D reconstruction at a synapse level, incorporating molecular, activity and morphodynamic information. LIONESS opens up avenues for studying the dynamic functional (nano-)architecture of living brain tissue.
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