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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Autores principales:	Giuseppe Vicidomini, Andreas Schönle, Haisen Ta, Kyu Young Han, Gael Moneron, Christian Eggeling, Stefan W Hell
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/42ae1630089d412aa3fe200f43173701
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id	oai:doaj.org-article:42ae1630089d412aa3fe200f43173701
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spelling	oai:doaj.org-article:42ae1630089d412aa3fe200f431737012021-11-18T08:00:51ZSTED nanoscopy with time-gated detection: theoretical and experimental aspects.1932-620310.1371/journal.pone.0054421https://doaj.org/article/42ae1630089d412aa3fe200f431737012013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23349884/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203In 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.Giuseppe VicidominiAndreas SchönleHaisen TaKyu Young HanGael MoneronChristian EggelingStefan W HellPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 1, p e54421 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q 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.
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.
format	article
author	Giuseppe Vicidomini Andreas Schönle Haisen Ta Kyu Young Han Gael Moneron Christian Eggeling Stefan W Hell
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)
publishDate	2013
url	https://doaj.org/article/42ae1630089d412aa3fe200f43173701
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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STED nanoscopy with time-gated detection: theoretical and experimental aspects.

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