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Multicolour Multilevel STED nanoscopy of Actin/Spectrin Organization at Synapses.

ABSTRACT: Superresolution fluorescence microscopy of multiple fluorophores still requires development. Here we present simultaneous three-colour stimulated emission depletion (STED) nanoscopy relying on a single STED beam at 620?nm. Toggling the STED beam between two or more power levels ("multilevelSTED") optimizes resolution and contrast in all colour channels, which are intrinsically co-aligned and well separated. Three-colour recording is demonstrated by imaging the nanoscale cytoskeletal organization in cultured hippocampal neurons. The down to ~35?nm resolution identified periodic actin/betaII spectrin lattices along dendrites and spines; however, at presynaptic and postsynaptic sites, these patterns were found to be absent. Both our multicolour scheme and the 620?nm STED line should be attractive for routine STED microscopy applications.

SUBMITTER: Sidenstein SC

PROVIDER: S-EPMC4879624 | biostudies-literature | 2016 May

REPOSITORIES: biostudies-literature

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Multicolour Multilevel STED nanoscopy of Actin/Spectrin Organization at Synapses.

Sidenstein Sven C SC D'Este Elisa E Böhm Marvin J MJ Danzl Johann G JG Belov Vladimir N VN Hell Stefan W SW

Scientific reports 20160525

Superresolution fluorescence microscopy of multiple fluorophores still requires development. Here we present simultaneous three-colour stimulated emission depletion (STED) nanoscopy relying on a single STED beam at 620 nm. Toggling the STED beam between two or more power levels ("multilevelSTED") optimizes resolution and contrast in all colour channels, which are intrinsically co-aligned and well separated. Three-colour recording is demonstrated by imaging the nanoscale cytoskeletal organization ...[more]

PMID: 27220554

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