STED nanoscopy with time-gated detection: theoretical and experimental aspects.
In a stimulated emission depletion (STED) microscope the region in which fluorescence markers can emit spontaneously shrinks with continued STED beam action after a singular excitation event. This fact has been recently used to substantially improve the effective spatial resolution in STED nanoscopy...
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doaj-42ae1630089d412aa3fe200f431737012021-03-03T20:25:17ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0181e5442110.1371/journal.pone.0054421STED nanoscopy with time-gated detection: theoretical and experimental aspects.Giuseppe VicidominiAndreas SchönleHaisen TaKyu Young HanGael MoneronChristian EggelingStefan W HellIn a stimulated emission depletion (STED) microscope the region in which fluorescence markers can emit spontaneously shrinks with continued STED beam action after a singular excitation event. This fact has been recently used to substantially improve the effective spatial resolution in STED nanoscopy using time-gated detection, pulsed excitation and continuous wave (CW) STED beams. We present a theoretical framework and experimental data that characterize the time evolution of the effective point-spread-function of a STED microscope and illustrate the physical basis, the benefits, and the limitations of time-gated detection both for CW and pulsed STED lasers. While gating hardly improves the effective resolution in the all-pulsed modality, in the CW-STED modality gating strongly suppresses low spatial frequencies in the image. Gated CW-STED nanoscopy is in essence limited (only) by the reduction of the signal that is associated with gating. Time-gated detection also reduces/suppresses the influence of local variations of the fluorescence lifetime on STED microscopy resolution.https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23349884/pdf/?tool=EBI |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Giuseppe Vicidomini Andreas Schönle Haisen Ta Kyu Young Han Gael Moneron Christian Eggeling Stefan W Hell |
spellingShingle |
Giuseppe Vicidomini Andreas Schönle Haisen Ta Kyu Young Han Gael Moneron Christian Eggeling Stefan W Hell STED nanoscopy with time-gated detection: theoretical and experimental aspects. PLoS ONE |
author_facet |
Giuseppe Vicidomini Andreas Schönle Haisen Ta Kyu Young Han Gael Moneron Christian Eggeling Stefan W Hell |
author_sort |
Giuseppe Vicidomini |
title |
STED nanoscopy with time-gated detection: theoretical and experimental aspects. |
title_short |
STED nanoscopy with time-gated detection: theoretical and experimental aspects. |
title_full |
STED nanoscopy with time-gated detection: theoretical and experimental aspects. |
title_fullStr |
STED nanoscopy with time-gated detection: theoretical and experimental aspects. |
title_full_unstemmed |
STED nanoscopy with time-gated detection: theoretical and experimental aspects. |
title_sort |
sted nanoscopy with time-gated detection: theoretical and experimental aspects. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2013-01-01 |
description |
In a stimulated emission depletion (STED) microscope the region in which fluorescence markers can emit spontaneously shrinks with continued STED beam action after a singular excitation event. This fact has been recently used to substantially improve the effective spatial resolution in STED nanoscopy using time-gated detection, pulsed excitation and continuous wave (CW) STED beams. We present a theoretical framework and experimental data that characterize the time evolution of the effective point-spread-function of a STED microscope and illustrate the physical basis, the benefits, and the limitations of time-gated detection both for CW and pulsed STED lasers. While gating hardly improves the effective resolution in the all-pulsed modality, in the CW-STED modality gating strongly suppresses low spatial frequencies in the image. Gated CW-STED nanoscopy is in essence limited (only) by the reduction of the signal that is associated with gating. Time-gated detection also reduces/suppresses the influence of local variations of the fluorescence lifetime on STED microscopy resolution. |
url |
https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23349884/pdf/?tool=EBI |
work_keys_str_mv |
AT giuseppevicidomini stednanoscopywithtimegateddetectiontheoreticalandexperimentalaspects AT andreasschonle stednanoscopywithtimegateddetectiontheoreticalandexperimentalaspects AT haisenta stednanoscopywithtimegateddetectiontheoreticalandexperimentalaspects AT kyuyounghan stednanoscopywithtimegateddetectiontheoreticalandexperimentalaspects AT gaelmoneron stednanoscopywithtimegateddetectiontheoreticalandexperimentalaspects AT christianeggeling stednanoscopywithtimegateddetectiontheoreticalandexperimentalaspects AT stefanwhell stednanoscopywithtimegateddetectiontheoreticalandexperimentalaspects |
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