4.8 Article

Yeast Replicator: A High-Throughput Multiplexed Microfluidics Platform for Automated Measurements of Single-Cell Aging

Journal

CELL REPORTS
Volume 13, Issue 3, Pages 634-644

Publisher

CELL PRESS
DOI: 10.1016/j.celrep.2015.09.012

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Funding

  1. New Scholar in Aging Award (Ellison Medical Foundation) [AG-NS-1015-13]
  2. NIH Director's New Innovator Award (NIH Director's Office)
  3. Gruber Science Fellowship
  4. NIH Genetics Predoctoral Research Training Program [T32 GM007499]
  5. NIH Director's New Innovator Award (National Institute on Aging) [1DP2AG050461-01]

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The yeast Saccharomyces cerevisiae is a model organism for replicative aging studies; however, conventional lifespan measurement platforms have several limitations. Here, we present a microfluidics platform that facilitates simultaneous lifespan and gene expression measurements of aging yeast cells. Our multiplexed high-throughput platform offers the capability to perform independent lifespan experiments using different yeast strains or growth media. Using this platform in minimal media environments containing glucose, we measured the full lifespan of individual yeast cells in wild-type and canonical gene deletion backgrounds. Compared to glucose, in galactose we observed a 16.8% decrease in replicative lifespan accompanied by an similar to 2-fold increase in single-cell oxidative stress levels reported by P-SOD1-mCherry. Using P-GAL1-YFP to measure the activity of the bistable galactose network, we saw that OFF and ON cells are similar in their lifespan. Our work shows that aging cells are committed to a single phenotypic state throughout their lifespan.

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