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Electrostatic Interactions of Fluorescent Molecules with Dielectric Interfaces Studied by 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.
KTH, School of Engineering Sciences (SCI), Applied Physics, Experimental Biomolecular Physics.ORCID iD: 0000-0003-3200-0374
2010 (English)In: International Journal of Molecular Sciences, ISSN 1422-0067, Vol. 11, no 2, 368-406 p.Article in journal (Refereed) Published
Abstract [en]

Electrostatic interactions between dielectric surfaces and different fluorophoresused in ultrasensitive fluorescence microscopy are investigated using objective-based TotalInternal Reflection Fluorescence Correlation Spectroscopy (TIR-FCS). The interfacialdynamics of cationic rhodamine 123 and rhodamine 6G, anionic/dianionic fluorescein,zwitterionic rhodamine 110 and neutral ATTO 488 are monitored at various ionic strengthsat physiological pH. As analyzed by means of the amplitude and time-evolution of theautocorrelation function, the fluorescent molecules experience electrostatic attraction orrepulsion at the glass surface depending on their charges. Influences of the electrostaticinteractions are also monitored through the triplet-state population and triplet relaxationtime, including the amount of detected fluorescence or the count-rate-per-moleculeparameter. These TIR-FCS results provide an increased understanding of how fluorophoresare influenced by the microenvironment of a glass surface, and show a promising approachfor characterizing electrostatic interactions at interfaces.

Place, publisher, year, edition, pages
MDPI , 2010. Vol. 11, no 2, 368-406 p.
Keyword [en]
electrostatic interactions, fluorescent molecules, glass interfaces, Total Internal Reflection, Fluorescence Correlation Spectroscopy
National Category
Physical Chemistry
Identifiers
URN: urn:nbn:se:kth:diva-12340DOI: 10.3390/ijms11020386ISI: 000274973500001Scopus ID: 2-s2.0-77649207561OAI: oai:DiVA.org:kth-12340DiVA: diva2:309801
Funder
Swedish Research Council, VR-621-2006-3197Knut and Alice Wallenberg Foundation
Note
QC 20100617Available from: 2010-04-12 Created: 2010-04-08 Last updated: 2011-01-20Bibliographically 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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