Abstract:
:Unraveling the genetic and epigenetic determinants of phenotypes is critical for understanding and re-engineering biology and would benefit from improved methods to separate cells based on phenotypes. Here, we report SPOTlight, a versatile high-throughput technique to isolate individual yeast or human cells with unique spatiotemporal profiles from heterogeneous populations. SPOTlight relies on imaging visual phenotypes by microscopy, precise optical tagging of single target cells, and retrieval of tagged cells by fluorescence-activated cell sorting. To illustrate SPOTlight's ability to screen cells based on temporal properties, we chose to develop a photostable yellow fluorescent protein for extended imaging experiments. We screened 3 million cells expressing mutagenesis libraries and identified a bright new variant, mGold, that is the most photostable yellow fluorescent protein reported to date. We anticipate that the versatility of SPOTlight will facilitate its deployment to decipher the rules of life, understand diseases, and engineer new molecules and cells.
journal_name
Sci Advjournal_title
Science advancesauthors
Lee J,Liu Z,Suzuki PH,Ahrens JF,Lai S,Lu X,Guan S,St-Pierre Fdoi
10.1126/sciadv.abb7438subject
Has Abstractpub_date
2020-10-23 00:00:00issue
43issn
2375-2548pii
6/43/eabb7438journal_volume
6pub_type
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