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dc.contributor.authorLuro, Scott
dc.contributor.authorPotvin-Trottier, Laurent
dc.contributor.authorOkumus, Burak
dc.contributor.authorPaulsson, Johan
dc.date.accessioned2022-08-25T16:54:00Z
dc.date.issued2019-11-25
dc.identifier.citationLuro, Scott, Laurent Potvin-Trottier, Burak Okumus, and Johan Paulsson. 2020. Isolating Live Cells after High-throughput, Long-term, Time-lapse Microscopy. Nature Methods 17, no. 1: 93-100.en_US
dc.identifier.issn1548-7091en_US
dc.identifier.issn1548-7105en_US
dc.identifier.urihttps://nrs.harvard.edu/URN-3:HUL.INSTREPOS:37373059*
dc.description.abstractSingle-cell genetic screens can be incredibly powerful, but current high-throughput platforms do not track dynamics, and even for non-dynamic properties they struggle to separate mutants of interest from phenotypic outliers of the wild-type population. Here we introduce SIFT, Single-cell Isolation Following Time-lapse imaging, to address these limitations. After imaging and tracking individual bacteria for tens of consecutive generations under tightly controlled growth conditions, cells of interest are isolated and propagated for downstream analysis, free of contamination and without genetic or physiological perturbations. The initial platform can characterize tens of thousands of cell lineages per day, making it possible to accurately screen complex phenotypes without the need for barcoding or genetic modifications. We applied SIFT to identify a set of ultraprecise synthetic gene oscillators, with mutants spanning a 30-fold range of average periods. This revealed novel design principles in synthetic biology and demonstrated the power of SIFT to reliably screen diverse dynamic phenotypes.en_US
dc.language.isoen_USen_US
dc.publisherSpringer Science and Business Media LLCen_US
dc.relationNature Methodsen_US
dash.licenseMETA_ONLY
dc.subjectBiotechnologyen_US
dc.subjectCell Biologyen_US
dc.subjectBiochemistryen_US
dc.subjectMolecular Biologyen_US
dc.titleIsolating Live Cells After High-Throughput, Long-Term, Time-Lapse Microscopyen_US
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden_US
dc.relation.journalNature Methodsen_US
dash.waiver2019-09-17
dc.date.available2022-08-25T16:54:00Z
dash.affiliation.otherHarvard Medical Schoolen_US
dc.identifier.doi10.1038/s41592-019-0620-7
dc.source.journalNat Methods
dash.source.volume17;1en_US
dash.source.page93-100en_US
dash.contributor.affiliatedLuro, Scott
dash.contributor.affiliatedPaulsson, Johan


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