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Triplet-State Investigations of Fluorescent Dyes at Dielectric Interfaces Using Total Internal Reflection Fluorescence Correlation Spectroscopy
KTH, School of Engineering Sciences (SCI), Applied Physics, Experimental Biomolecular Physics.ORCID iD: 0000-0002-5584-9170
KTH, School of Engineering Sciences (SCI), Applied Physics, Experimental Biomolecular Physics.
KTH, School of Engineering Sciences (SCI), Applied Physics, Experimental Biomolecular Physics.
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2009 (English)In: Journal of Physical Chemistry A, ISSN 1089-5639, E-ISSN 1520-5215, Vol. 113, no 19, 5554-5566 p.Article in journal (Refereed) Published
Abstract [en]

The triplet-state kinetics of several fluorescent dyes used in ultrasensitive fluorescence microscopy are investigated using total internal reflection fluorescence correlation spectroscopy (TIR-FCS). A theoretical outline of the correlation analysis and the physical aspects of evanescent excitation and fluorescence emission at dielectric interfaces are given. From this analysis, the rates of intersystem crossing and triplet decay are deduced for fluorescein, ATTO 488, rhodamine 110, rhodamine 123, and rhodamine 6G in aqueous buffer solutions. All investigated dyes show slightly higher triplet rates at the dielectric interface compared to bulk solution measurements. We attribute this enhancement to possible modifications of the dyes’ photophysical properties near a dielectric interface. In the case of rhodamine 6G, the impact of changes in the dye concentration, ionic strength of the solvent, and potassium iodide concentration are also investigated. This leads to a better understanding of the influences of dye−dye, dye−solvent, and dye−surface interactions on the increased triplet intersystem crossing and triplet decay rates. The study shows that analysis of triplet-state kinetics by TIR-FCS not only results in a better understanding of how the photophysical properties of the dyes are affected by the presence of an interface, but also provides a means for probing the microenvironment near dielectric interfaces.

Place, publisher, year, edition, pages
Washington, DC: American Chemical Society , 2009. Vol. 113, no 19, 5554-5566 p.
Keyword [en]
fluorescence correlation spectroscopy, FCS, triplet state, surface effects, TIR, TIR-FCS, total internal reflection
National Category
Physical Chemistry
Identifiers
URN: urn:nbn:se:kth:diva-12338DOI: 10.1021/jp8110088ISI: 000265887700010Scopus ID: 2-s2.0-66149146666OAI: oai:DiVA.org:kth-12338DiVA: diva2:309782
Note
QC 20110208Available 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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Blom, Hans

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