---
_id: '6808'
abstract:
- lang: eng
text: Super-resolution fluorescence microscopy has become an important catalyst
for discovery in the life sciences. In STimulated Emission Depletion (STED) microscopy,
a pattern of light drives fluorophores from a signal-emitting on-state to a non-signalling
off-state. Only emitters residing in a sub-diffraction volume around an intensity
minimum are allowed to fluoresce, rendering them distinguishable from the nearby,
but dark fluorophores. STED routinely achieves resolution in the few tens of nanometers
range in biological samples and is suitable for live imaging. Here, we review
the working principle of STED and provide general guidelines for successful STED
imaging. The strive for ever higher resolution comes at the cost of increased
light burden. We discuss techniques to reduce light exposure and mitigate its
detrimental effects on the specimen. These include specialized illumination strategies
as well as protecting fluorophores from photobleaching mediated by high-intensity
STED light. This opens up the prospect of volumetric imaging in living cells and
tissues with diffraction-unlimited resolution in all three spatial dimensions.
article_processing_charge: No
article_type: original
author:
- first_name: Wiebke
full_name: Jahr, Wiebke
id: 425C1CE8-F248-11E8-B48F-1D18A9856A87
last_name: Jahr
- first_name: Philipp
full_name: Velicky, Philipp
id: 39BDC62C-F248-11E8-B48F-1D18A9856A87
last_name: Velicky
orcid: 0000-0002-2340-7431
- first_name: Johann G
full_name: Danzl, Johann G
id: 42EFD3B6-F248-11E8-B48F-1D18A9856A87
last_name: Danzl
orcid: 0000-0001-8559-3973
citation:
ama: Jahr W, Velicky P, Danzl JG. Strategies to maximize performance in STimulated
Emission Depletion (STED) nanoscopy of biological specimens. Methods. 2020;174(3):27-41.
doi:10.1016/j.ymeth.2019.07.019
apa: Jahr, W., Velicky, P., & Danzl, J. G. (2020). Strategies to maximize performance
in STimulated Emission Depletion (STED) nanoscopy of biological specimens. Methods.
Elsevier. https://doi.org/10.1016/j.ymeth.2019.07.019
chicago: Jahr, Wiebke, Philipp Velicky, and Johann G Danzl. “Strategies to Maximize
Performance in STimulated Emission Depletion (STED) Nanoscopy of Biological Specimens.”
Methods. Elsevier, 2020. https://doi.org/10.1016/j.ymeth.2019.07.019.
ieee: W. Jahr, P. Velicky, and J. G. Danzl, “Strategies to maximize performance
in STimulated Emission Depletion (STED) nanoscopy of biological specimens,” Methods,
vol. 174, no. 3. Elsevier, pp. 27–41, 2020.
ista: Jahr W, Velicky P, Danzl JG. 2020. Strategies to maximize performance in STimulated
Emission Depletion (STED) nanoscopy of biological specimens. Methods. 174(3),
27–41.
mla: Jahr, Wiebke, et al. “Strategies to Maximize Performance in STimulated Emission
Depletion (STED) Nanoscopy of Biological Specimens.” Methods, vol. 174,
no. 3, Elsevier, 2020, pp. 27–41, doi:10.1016/j.ymeth.2019.07.019.
short: W. Jahr, P. Velicky, J.G. Danzl, Methods 174 (2020) 27–41.
date_created: 2019-08-12T16:36:32Z
date_published: 2020-03-01T00:00:00Z
date_updated: 2023-08-17T13:59:57Z
day: '01'
department:
- _id: JoDa
doi: 10.1016/j.ymeth.2019.07.019
external_id:
isi:
- '000525860400005'
pmid:
- '31344404'
intvolume: ' 174'
isi: 1
issue: '3'
language:
- iso: eng
main_file_link:
- open_access: '1'
url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7100895/
month: '03'
oa: 1
oa_version: Submitted Version
page: 27-41
pmid: 1
project:
- _id: 265CB4D0-B435-11E9-9278-68D0E5697425
call_identifier: FWF
grant_number: I03600
name: Optical control of synaptic function via adhesion molecules
- _id: 2668BFA0-B435-11E9-9278-68D0E5697425
grant_number: LT00057
name: High-speed 3D-nanoscopy to study the role of adhesion during 3D cell migration
publication: Methods
publication_identifier:
issn:
- 1046-2023
publication_status: published
publisher: Elsevier
quality_controlled: '1'
scopus_import: '1'
status: public
title: Strategies to maximize performance in STimulated Emission Depletion (STED)
nanoscopy of biological specimens
type: journal_article
user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8
volume: 174
year: '2020'
...