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Measuring Single-Cell Phenotypic Growth Heterogeneity Using a Microfluidic Cell Volume Sensor.

Wenyang Jing1, Brendan Camellato2, Ian J Roney2

  • 1Department of Physics, University of Ottawa, Ottawa, Ontario, Canada.

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|December 15, 2018
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Summary
This summary is machine-generated.

Single yeast cells were studied using a microfluidic sensor to measure growth. High PDR5 gene expression improved survival in the antibiotic cycloheximide, while low expression was better in rich media.

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Microfluidics

Background:

  • The PDR5 gene in Saccharomyces cerevisiae encodes a membrane transporter conferring multidrug resistance.
  • Phenotypic variation in PDR5 expression impacts cellular fitness and response to cytotoxic compounds.
  • Understanding single-cell responses is crucial for heterogeneous population studies.

Purpose of the Study:

  • To evaluate the volumetric growth rate and fitness of single yeast cells with differing PDR5 expression levels.
  • To assess cellular response to rich media versus media containing the antibiotic cycloheximide.
  • To demonstrate the utility of an imaging-integrated microfluidic sensor for high-resolution single-cell analysis.

Main Methods:

  • Utilized an imaging-integrated microfluidic cell volume sensor for real-time growth rate measurement.
  • Isolated and trapped single Saccharomyces cerevisiae cells using fluorescent markers.
  • Measured single-cell growth rates in two distinct on-chip environments: rich media and cycloheximide-dosed media.

Main Results:

  • Cells with low PDR5 expression exhibited higher fitness in rich media.
  • Cells with high PDR5 expression demonstrated superior fitness in the presence of cycloheximide.
  • Cycloheximide significantly reduced the fitness of low PDR5-expressing cells, with minimal impact on high PDR5-expressing cells.

Conclusions:

  • The imaging-integrated microfluidic sensor enables high-resolution, single-cell analysis of growth and fitness.
  • PDR5 expression level is a key determinant of yeast cell fitness in the presence of specific drugs.
  • This technology is applicable for characterizing heterogeneous cell populations under various conditions.