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Studying ion exchange in solution and at biological membranes by FCS
KTH, School of Engineering Sciences (SCI), Applied Physics, Experimental Biomolecular Physics.
2013 (English)In: Fluorescence Fluctuation Spectroscopy (FFS), Part B / [ed] Sergei Y Tetin, Elsevier, 2013, 231-252 p.Chapter in book (Refereed)
Abstract [en]

By FCS, a wide range of processes can be studied, covering time ranges from subnanoseconds to seconds. In principle, any process at equilibrium conditions can be measured, which reflects itself by a change in the detected fluorescence intensity. In this review, it is described how FCS and variants thereof can be used to monitor ion exchange, in solution and along biological membranes. Analyzing fluorescence fluctuations of ion-sensitive fluorophores by FCS offers selective advantages over other techniques for measuring local ion concentrations, and, in particular, for studying exchange kinetics of ions on a very local scale. This opens for several areas of application. The FCS approach was used to investigate fundamental aspects of proton exchange at and along biological membranes. The protonation relaxation rate, as measured by FCS for a pH-sensitive dye, can also provide information about local accessibility/ interaction of a particular labeling site and conformational states of biomolecules, in a similar fashion as in a fluorescence quenching experiment. The same FCS concept can also be applied to ion exchange studies using other ion-sensitive fluorophores, and by use of dyes sensitive to other ambient conditions the concept can be extended also beyond ion exchange studies.

Place, publisher, year, edition, pages
Elsevier, 2013. 231-252 p.
, Methods in Enzymology, ISSN 0076-6879 ; 519
Keyword [en]
Biomembrane, Buffer, Fluorescence correlation spectroscopy, pH, Protonation
National Category
Biochemistry and Molecular Biology Biophysics
Research subject
Biological Physics
URN: urn:nbn:se:kth:diva-118216DOI: 10.1016/B978-0-12-405539-1.00008-7ISI: 000318528200008ScopusID: 2-s2.0-84872008594ISBN: 978-012405539-1OAI: diva2:605058

QC 20130213

Available from: 2013-02-13 Created: 2013-02-13 Last updated: 2016-03-10Bibliographically approved

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Widengren Meth Enzymol 2013(639 kB)14 downloads
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