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Journal Article

Multicolour multilevel STED nanoscopy of actin/spectrin organization at synapses.

MPS-Authors
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Sidenstein,  S.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Böhm,  M. J.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Danzl,  J. G.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Belov,  V. N.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

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Hell,  S. W.
Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society;

External Ressource
Fulltext (public)

2305864.pdf
(Publisher version), 2MB

Supplementary Material (public)

2305864_Suppl.pdf
(Supplementary material), 11MB

Citation

Sidenstein, S., D’Este, E., Böhm, M. J., Danzl, J. G., Belov, V. N., & Hell, S. W. (2016). Multicolour multilevel STED nanoscopy of actin/spectrin organization at synapses. Scientific Reports, 6: 26725. doi:10.1038/srep26725.


Cite as: http://hdl.handle.net/11858/00-001M-0000-002A-E3E6-0
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.