Title |
Use of Fluorescence Lifetime Imaging Microscopy (FLIM) as a Timer of Cell Cycle S Phase
|
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Published in |
PLOS ONE, December 2016
|
DOI | 10.1371/journal.pone.0167385 |
Pubmed ID | |
Authors |
Irina A. Okkelman, Ruslan I. Dmitriev, Tara Foley, Dmitri B. Papkovsky |
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. |
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Other | 0 | 0% |
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Demographic breakdown
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Members of the public | 7 | 32% |
Practitioners (doctors, other healthcare professionals) | 1 | 5% |
Science communicators (journalists, bloggers, editors) | 1 | 5% |
Mendeley readers
Geographical breakdown
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Unknown | 52 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Master | 10 | 19% |
Other | 6 | 12% |
Researcher | 6 | 12% |
Student > Postgraduate | 3 | 6% |
Other | 5 | 10% |
Unknown | 11 | 21% |
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Medicine and Dentistry | 2 | 4% |
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