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Transient state imaging of live cells using single plane illumination and arbitrary duty cycle excitation pulse trains
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
2015 (English)In: Journal of Biophotonics, ISSN 1864-063X, E-ISSN 1864-0648, Vol. 8, no 5, 392-400 p.Article in journal (Refereed) Published
Abstract [en]

We demonstrate the applicability of Single Plane Illumination Microscopy to Transient State Imaging (TRAST), offering sensitive microenvironmental information together with optical sectioning and reduced overall excitation light exposure of the specimen. The concept is verified by showing that transition rates can be determined accurately for free dye in solution and that fluorophore transition rates can be resolved pixel-wise in live cells. Furthermore, we derive a new theoretical framework for analyzing TRAST data acquired with arbitrary duty cycle pulse trains. By this analysis it is possible to reduce the overall measurement time and thereby enhance the frame rates in TRAST imaging.

Place, publisher, year, edition, pages
Wiley-VCH Verlagsgesellschaft, 2015. Vol. 8, no 5, 392-400 p.
Keyword [en]
fluorescence, single plane illumination microscopy, transient state imaging, triplet state, redox state, live cell imaging
National Category
Biophysics
Research subject
Biological Physics
Identifiers
URN: urn:nbn:se:kth:diva-170699DOI: 10.1002/jbio.201400015ISI: 000355983800004PubMedID: 24706633Scopus ID: 2-s2.0-84929510753OAI: oai:DiVA.org:kth-170699DiVA: diva2:840133
Funder
EU, European Research Council, FLUODIAMON 201 837Swedish Research Council, VRNT 2012-3045Knut and Alice Wallenberg Foundation, KAW 2011.0218
Note

QC 20150707

Available from: 2015-07-07 Created: 2015-07-03 Last updated: 2017-12-04Bibliographically approved

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