Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Modulated or alternating excitation in fluorescence correlation spectroscopy
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.
KTH, School of Engineering Sciences (SCI), Applied Physics, Experimental Biomolecular Physics.ORCID iD: 0000-0003-3200-0374
2009 (English)In: SINGLE MOLECULE SPECTROSCOPY AND IMAGING / [ed] Enderlein J; Gryczynski ZK; Erdmann R, 2009, Vol. 7185Conference paper, Published paper (Refereed)
Abstract [en]

We have previously shown that formation of triplet states and other photo-induced states can be controlled by modulating the excitation with pulse widths and periods in the range of the transition times of the involved states. However, modulating the excitation in fluorescence correlation spectroscopy (FCS) measurements normally destroys correlation information and induces ringing in the correlation curve. We have introduced and experimentally verified a method to retrieve the full correlation curves from FCS measurements with modulated excitation and arbitrarily low fraction of active excitation. Modulated excitation applied to FCS experiments was shown to suppress the triplet build-up more efficiently than reducing excitation power with continuous wave excitation. The usefulness of the method was demonstrated by measurements done on fluorescein at different pH, where suppression of the triplet significantly facilitates the analysis of the protonation kinetics. Using a fluorophore where the protonation-coupled fluorescence intensity fluctuations are due to spectral shifts, introduction of two-color alternating excitation and spectral cross-correlation can turn the protonation component of the correlation curve into an anti-correlation and further facilitate the distinction of this component from those of other processes.

Place, publisher, year, edition, pages
2009. Vol. 7185
Series
Proceedings of SPIE-The International Society for Optical Engineering, ISSN 0277-786X
Keyword [en]
fluorescence correlation spectroscopy, fluorescence cross-correlation spectroscopy, modulated excitation, alternating excitation, photobleaching, triplet state, protonation
National Category
Biophysics
Identifiers
URN: urn:nbn:se:kth:diva-30563DOI: 10.1117/12.808912ISI: 000285712600007Scopus ID: 2-s2.0-66649103005ISBN: 978-0-8194-7431-5 (print)OAI: oai:DiVA.org:kth-30563DiVA: diva2:401031
Conference
Conference on Single Molecule Spectroscopy and Imaging II San Jose, CA, JAN 24-25, 2009
Note
QC 20110301Available from: 2011-03-01 Created: 2011-02-28 Last updated: 2011-03-01Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full textScopus

Search in DiVA

By author/editor
Persson, GustavSandén, TorThyberg, PerWidengren, Jerker
By organisation
Experimental Biomolecular Physics
Biophysics

Search outside of DiVA

GoogleGoogle Scholar

doi
isbn
urn-nbn

Altmetric score

doi
isbn
urn-nbn
Total: 139 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf