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Characterization of new fluorescent labels for ultra-high resolution microscopy
KTH, School of Engineering Sciences (SCI), Applied Physics, Experimental Biomolecular Physics.
ATTO-TEC GmbH, Germany.
University of Siegen, Germany.
University of Siegen, Germany.
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2008 (English)In: Photochemical and Photobiological Sciences, ISSN 1474-905X, E-ISSN 1474-9092, Vol. 7, no 11, 1378-1385 p.Article in journal (Refereed) Published
Abstract [en]

Photo-induced switching of dyes into dark, long-lived states, such as a triplet state, has recently gained increasing interest, as a means to achieve ultra-high optical resolution. Additionally, these long lived states are often highly environment-sensitive and their photodynamics can thus offer additional independent fluorescence-based information. However, although providing a useful mechanism for photo-induced switching, the triplet state often appears as a precursor state for photobleaching, which potentially can limit its usefulness. In this work, a set of rhodamine and pyronin dyes, modified by substitution of heavy atoms and nitrogen within or close to the central xanthene unit of the dyes, were investigated with respect to their triplet state dynamics and photostabilities, under conditions relevant for ultra-high resolution microscopy. Out of the dyes investigated, in particular the rhodamine and pyronin dyes with a sulfur atom replacing the central oxygen atom in the xanthene unit were found to meet the requirements for ultra-high resolution microscopy, combining a prominent triplet state yield with reasonable photostability.

Place, publisher, year, edition, pages
2008. Vol. 7, no 11, 1378-1385 p.
Keyword [en]
fluorescence correlation spectroscopy, super resolution, tripet state, GSD
National Category
Physical Chemistry
Identifiers
URN: urn:nbn:se:kth:diva-12336DOI: 10.1039/b810991pISI: 000261733700003PubMedID: 18958325Scopus ID: 2-s2.0-55149095492OAI: oai:DiVA.org:kth-12336DiVA: diva2:309777
Note
QC 20100617Available from: 2010-04-12 Created: 2010-04-08 Last updated: 2017-12-12Bibliographically approved
In thesis
1. Photo-induced dark states influorescence spectroscopy – investigations & applications
Open this publication in new window or tab >>Photo-induced dark states influorescence spectroscopy – investigations & applications
2010 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

This thesis focuses on investigations of transient dark states of fluorescentmolecules using spectroscopic techniques. The main purpose is to show andconvince the reader that transient dark states are not always a nuisance, butalso represent an additional source of information. Several studies with fluorescencecorrelation spectroscopy were performed, all related to non-fluorescentstates such as triplet state or isomerized states.Photobleaching is one of the main problems in virtually all of the fluorescencetechniques. In this thesis, mechanisms that retard photobleaching arecharacterized. Several compounds, antioxidants and triplet state quenchers,which decrease photobleaching, are studied, and guidelines for achieving optimalfluorescence brightness using these compounds are presented.Triplet state quenching by several compounds was studied. Detailed investigationsof the fluorescence quencher potassium iodide demonstratedthat for some of fluorophores, except of quenching, there is fluorescence enhancementmechanism present. In agreement with the first publication inthis thesis, antioxidative properties were found to play an important role inthe fluorescence enhancement. Quenching of the triplet state is proposedas a tool for monitoring diffusion mediated reactions over a wide range offrequencies.Specially designed fluorophores combining high triplet yields with reasonablefluorescence brightness and photostability were characterized forpossible applications in novel super-resolution imaging techniques based onfluorescence photoswitching. Except of benefits for imaging techniques, photoinducedswitching to non-fluorescent states could be used for monitoringmolecular diffusion, which was also demonstrated in this thesis.Studies of the triplet state kinetics of fluorophores close to dielectric interfaceswere performed using fluorescence spectroscopy. The analysis of thetriplet state kinetic can provide information about the local microenvironmentand electrostatic interactions near dielectric interfaces.

Place, publisher, year, edition, pages
Stockholm: KTH, 2010. vii, 111 p.
Series
Trita-FYS, ISSN 0280-316X ; 2010:20
Keyword
fluorescence correlation spectroscopy, triplet state, isomerisation, photobleaching, quenching, diffusion, total internal reflection, interface
National Category
Physical Chemistry
Identifiers
urn:nbn:se:kth:diva-12372 (URN)978-91-628-8108-5 (ISBN)
Public defence
2010-04-29, FA31, Albanova University Center, Roslagstullsbacken 21, Stockholm, 13:00 (English)
Opponent
Supervisors
Note
QC 20100414Available from: 2010-04-14 Created: 2010-04-12 Last updated: 2011-01-20

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