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The effect of photoswitching kinetics and labeling densities on super-resolution fluorescence imaging
Author(s):
Sebastian van de Linde
,
Steve Wolter
,
Mike Heilemann
,
Markus Sauer
Publication date
Created:
September 15 2010
Publication date
(Print):
September 15 2010
Journal:
Journal of Biotechnology
Publisher:
Elsevier BV
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Abstract
Super-resolution fluorescence imaging methods based on reversible photoswitching of fluorophores with subsequent localization currently develop to promising tools for cellular imaging. Since most of these methods rely on the transfer of the majority of fluorophores to a non-fluorescent dark state and precise localization of separated fluorescent fluorophores, the photophysical properties of photoswitchable fluorophores have to be carefully controlled. The achievable resolution and herewith the ability to resolve a structural feature depends not only on the brightness of the fluorophores, but also on the labeling density and on the stability or lifetime of the non-fluorescent dark state. Here, we discuss how the ratio of off- and on-switching of a fluorophore affects resolution. We compare experimental data with theoretical simulations and present a strategy to customize photoswitching characteristics to achieve optimal optical resolution. Copyright © 2010 Elsevier B.V. All rights reserved.
Related collections
Journal of Applied Biology and Biotechnology
Author and article information
Journal
Title:
Journal of Biotechnology
Abbreviated Title:
Journal of Biotechnology
Publisher:
Elsevier BV
ISSN (Print):
01681656
Publication date Created:
September 15 2010
Publication date (Print):
September 15 2010
Volume
: 149
Issue
: 4
Pages
: 260-266
Article
DOI:
10.1016/j.jbiotec.2010.02.010
SO-VID:
cd0a0dde-d789-4e72-b3a5-0d5d11d51356
Copyright ©
© 2010
License:
https://www.elsevier.com/tdm/userlicense/1.0/
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