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

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dc.contributor.author Papkovsky, Dmitri B.
dc.contributor.author Okkelman, Irina A.
dc.contributor.author Dmitriev, Ruslan I.
dc.contributor.author Foley, Tara
dc.contributor.author Papkovsky, Dmitri B.
dc.date.accessioned 2017-01-06T15:32:49Z
dc.date.available 2017-01-06T15:32:49Z
dc.date.issued 2016-12-14
dc.identifier.citation Okkelman, 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.0167385 en
dc.identifier.volume 11 en
dc.identifier.issued 12 en
dc.identifier.startpage e0167385-1 en
dc.identifier.endpage e0167385-18 en
dc.identifier.isbn 10.1371/journal.pone.0167385
dc.identifier.issn 1932-6203
dc.identifier.uri http://hdl.handle.net/10468/3444
dc.description.abstract Incorporation 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.sponsorship Science Foundation of Ireland (SFI grants 13/SIRG/2144 (RID) and 12/RC/2276 (DBP, IAO)) en
dc.format.mimetype application/pdf en
dc.language.iso en en
dc.publisher Public Library of Science en
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.uri https://creativecommons.org/licenses/by/4.0/ en
dc.subject Organoids en
dc.subject Cell cycle and cell division en
dc.subject Cell staining en
dc.subject Fluorescence imaging en
dc.subject Synthesis phase en
dc.subject Gastrointestinal tract en
dc.subject Cell proliferation en
dc.subject Fluorescence microscopy en
dc.subject.lcsh Science Foundation Ireland en
dc.title Use of fluorescence lifetime imaging microscopy (FLIM) as a timer of cell cycle S phase en
dc.type Article (peer-reviewed) en
dc.internal.authorcontactother Ruslan Dmitriev, Biochemistry , University College Cork, Cork, Ireland. +353-21-490-3000 Email: r.dmitriev@ucc.ie en
dc.internal.availability Full text available en
dc.date.updated 2017-01-06T15:20:59Z
dc.description.version Published Version en
dc.internal.rssid 378454955
dc.description.status Peer reviewed en
dc.identifier.journaltitle Plos One en
dc.internal.copyrightchecked No !!CORA!! en
dc.internal.licenseacceptance Yes en
dc.internal.IRISemailaddress r.dmitriev@ucc.ie en


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© 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. Except where otherwise noted, this item's license is described as © 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.
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