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  STED nanoscopy with time-gated detection: Theoretical and experimental aspects.

Vicidomini, G., Schönle, A., Ta, H., Han, K. Y., Moneron, G., Eggeling, C., et al. (2013). STED nanoscopy with time-gated detection: Theoretical and experimental aspects. PLoS One, 8(1): e54421. doi:10.1371/journal.pone.0054421.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-000E-7743-A Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0027-C856-8
Genre: Journal Article

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Vicidomini, G.1, Author              
Schönle, A.1, Author              
Ta, H.1, Author              
Han, K. Y.1, Author              
Moneron, G.1, Author              
Eggeling, C.1, Author              
Hell, S. W.1, Author              
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1Department of NanoBiophotonics, MPI for biophysical chemistry, Max Planck Society, ou_578627              

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 Abstract: 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.

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Language(s): eng - English
 Dates: 2013-01-18
 Publication Status: Published online
 Pages: 12
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 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1371/journal.pone.0054421
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Title: PLoS One
Source Genre: Journal
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Pages: - Volume / Issue: 8 (1) Sequence Number: e54421 Start / End Page: - Identifier: ISSN: 1932-6203