Versatile Phenotype-Activated Cell Sorting

Authors

Jihwan Lee
Zhuohe Liu
Peter H Suzuki
John F Ahrens
Shujuan Lai
Xiaoyu Lu
Sihui Guan
François St-Pierre

Publication Date

10-1-2020

Journal

Science Advances

DOI

10.1126/sciadv.abb7438

PMID

33097540

PMCID

PMC7608836

PubMedCentral® Posted Date

10-23-2020

PubMedCentral® Full Text Version

Post-print

Published Open-Access

no

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.

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