Use of fluorescence lifetime imaging microscopy (FLIM) as a timer of cell cycle S phase

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dc.contributor.authorPapkovsky, Dmitri B.
dc.contributor.authorOkkelman, Irina A.
dc.contributor.authorDmitriev, Ruslan I.
dc.contributor.authorFoley, Tara
dc.contributor.authorPapkovsky, Dmitri B.
dc.date.accessioned2017-01-06T15:32:49Z
dc.date.available2017-01-06T15:32:49Z
dc.date.issued2016-12-14
dc.date.updated2017-01-06T15:20:59Z
dc.description.abstractIncorporation of thymidine analogues in replicating DNA, coupled with antibody and fluorophore staining, allows analysis of cell proliferation, but is currently limited to monolayer cultures, fixed cells and end-point assays. We describe a simple microscopy imaging method for live real-time analysis of cell proliferation, S phase progression over several division cycles, effects of anti-proliferative drugs and other applications. It is based on the prominent (~ 1.7-fold) quenching of fluorescence lifetime of a common cell-permeable nuclear stain, Hoechst 33342 upon the incorporation of 5-bromo-2’-deoxyuridine (BrdU) in genomic DNA and detection by fluorescence lifetime imaging microscopy (FLIM). We show that quantitative and accurate FLIM technique allows high-content, multi-parametric dynamic analyses, far superior to the intensity-based imaging. We demonstrate its uses with monolayer cell cultures, complex 3D tissue models of tumor cell spheroids and intestinal organoids, and in physiological study with metformin treatment.en
dc.description.sponsorshipScience Foundation of Ireland (SFI grants 13/SIRG/2144 (RID) and 12/RC/2276 (DBP, IAO))en
dc.description.statusPeer revieweden
dc.description.versionPublished Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationOkkelman, I. A., Dmitriev, R. I., Foley, T. and Papkovsky, D. B. (2016) 'Use of Fluorescence Lifetime Imaging Microscopy (FLIM) as a Timer of Cell Cycle S Phase', PLOS ONE, 11(12), pp. e0167385. doi:10.1371/journal.pone.0167385en
dc.identifier.endpagee0167385-18en
dc.identifier.isbn10.1371/journal.pone.0167385
dc.identifier.issn1932-6203
dc.identifier.issued12en
dc.identifier.journaltitlePlos Oneen
dc.identifier.startpagee0167385-1en
dc.identifier.urihttps://hdl.handle.net/10468/3444
dc.identifier.volume11en
dc.language.isoenen
dc.publisherPublic Library of Scienceen
dc.rights© 2016 Okkelman et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en
dc.subjectOrganoidsen
dc.subjectCell cycle and cell divisionen
dc.subjectCell stainingen
dc.subjectFluorescence imagingen
dc.subjectSynthesis phaseen
dc.subjectGastrointestinal tracten
dc.subjectCell proliferationen
dc.subjectFluorescence microscopyen
dc.subject.lcshScience Foundation Irelanden
dc.titleUse of fluorescence lifetime imaging microscopy (FLIM) as a timer of cell cycle S phaseen
dc.typeArticle (peer-reviewed)en
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