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Quantitative flow cytometry to understand population heterogeneity in response to changes in substrate availability in escherichia coli and saccharomyces cerevisiae chemostats
Citation Link: https://doi.org/10.15480/882.8164
Publikationstyp
Journal Article
Date Issued
2019-08-05
Sprache
English
Author(s)
Department of Chemical and Biochemical Engineering, Technical University of Denmark, Lyngby, Denmark
Johanson, Ted
Han, Shanshan
Carlquist, Magnus
Lantz, Anna Eliasson
TORE-DOI
Volume
7
Issue
AUG
Start Page
1
End Page
15
Article Number
187
Citation
Frontiers in Bioengineering and Biotechnology 7 (AUG): 1 - 15, 187 (2019-08-05)
Publisher DOI
Scopus ID
Publisher
Frontiers Media S.A.
Microbial cells in bioprocesses are usually described with averaged parameters. But in fact, single cells within populations vary greatly in characteristics such as stress resistance, especially in response to carbon source gradients. Our aim was to introduce tools to quantify population heterogeneity in bioprocesses using a combination of reporter strains, flow cytometry, and easily comprehensible parameters. We calculated mean, mode, peak width, and coefficient of variance to describe distribution characteristics and temporal shifts in fluorescence intensity. The skewness and the slope of cumulative distribution function plots illustrated differences in distribution shape. These parameters are person-independent and precise. We demonstrated this by quantifying growth-related population heterogeneity of Saccharomyces cerevisiae and Escherichia coli reporter strains in steady-state of aerobic glucose-limited chemostat cultures at different dilution rates and in response to glucose pulses. Generally, slow-growing cells showed stronger responses to glucose excess than fast-growing cells. Cell robustness, measured as membrane integrity after exposure to freeze-thaw treatment, of fast-growing cells was strongly affected in subpopulations of low membrane robustness. Glucose pulses protected subpopulations of fast-growing but not slower-growing yeast cells against membrane damage. Our parameters could successfully describe population heterogeneity, thereby revealing physiological characteristics that might have been overlooked during traditional averaged analysis.
Subjects
Flow cytometry
Glucose pulse
Membrane robustness
Population heterogeneity
Quantitative flow cytometry
Reporter strain
DDC Class
570: Life Sciences, Biology
Publication version
publishedVersion
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