Define flow cytometry8/16/2023 Optimization of industrial cultivation processes requires comprehensive analysis and understanding of host strain physiology throughout the cultivation. Our parameters could successfully describe population heterogeneity, thereby revealing physiological characteristics that might have been overlooked during traditional averaged analysis. Glucose pulses protected subpopulations of fast-growing but not slower-growing yeast cells against membrane damage. 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. Generally, slow-growing cells showed stronger responses to glucose excess than fast-growing cells. 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. These parameters are person-independent and precise. The skewness and the slope of cumulative distribution function plots illustrated differences in distribution shape. We calculated mean, mode, peak width, and coefficient of variance to describe distribution characteristics and temporal shifts in fluorescence intensity. Our aim was to introduce tools to quantify population heterogeneity in bioprocesses using a combination of reporter strains, flow cytometry, and easily comprehensible parameters. But in fact, single cells within populations vary greatly in characteristics such as stress resistance, especially in response to carbon source gradients. Microbial cells in bioprocesses are usually described with averaged parameters. 4Division of Applied Microbiology, Department of Chemistry, Lund University, Lund, Sweden.3Section of Microbiology, Department of Biology, University of Copenhagen, Copenhagen, Denmark.1Department of Chemical and Biochemical Engineering, Technical University of Denmark, Lyngby, Denmark.Anna-Lena Heins 1*†, Ted Johanson 2, Shanshan Han 3, Luisa Lundin 3 †, Magnus Carlquist 4, Krist V.
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