Dynamic single-cell analysis of Saccharomyces cerevisiae under process perturbation: comparison of different methods for monitoring the intensity of population heterogeneity
[en] BACKGROUND: Single cell biology has attracted a lot of attention these past few years and has led to numerous fundamental results pointing out the heterogeneity of clonal cell populations. In this context, microbial phenotypic heterogeneity under bioprocessing conditions needs to be further investigated. In this study, yeast based processes have been investigated by using on-line flow cytometry in combination with a fluorescent transcriptional reporter (GFP) and viability fluorescence tags (propidium iodide, PI). Methods aiming at expressing the dispersion of these fluorescence tags among the yeast populations have been investigated for different bioreactor operating conditions.
RESULTS: Yeast viability was determined on the basis of PI uptake. Segregation between PI negative and positive subpopulations could be efficiently quantified on the basis of the mean-to-median ratio or the amplitude of the interquartile range. On the other hand, the same quantification could not be made for the segregation occurring at the level of GFP synthesis. Indeed, when cells were exposed to sub-lethal or mild stresses (such as in scale-down reactors) two GFP subpopulations could be visualized by real-time FC, but quantification by one of the above-mentioned methods was not possible.
CONCLUSIONS: Yeast population heterogeneity was observed in representative bioreactor operating conditions. Difficulties for the determination of segregation at the level of GFP synthesis point out the fact that one needs to understand the segregation mechanisms for the applied fluorescent reporters, to judge whether simple mathematical tools may be applied or if more sophisticated computational tools are needed for the quantification of the microbial population segregation.
Disciplines :
Biotechnology
Author, co-author :
Delvigne, Frank ; Université de Liège - ULiège > Chimie et bio-industries > Bio-industries
Baert, Jonathan ; Université de Liège - ULiège > Chimie et bio-industries > Bio-industries
Gofflot, Sébastien
Lejeune, Annick ; Université de Liège - ULiège > Chimie et bio-industries > Bio-industries
Telek, Samuel ; Université de Liège - ULiège > Chimie et bio-industries > Bio-industries
Johanson, Ted
Lantz, Anna Eliasson
Language :
English
Title :
Dynamic single-cell analysis of Saccharomyces cerevisiae under process perturbation: comparison of different methods for monitoring the intensity of population heterogeneity
Publication date :
2015
Journal title :
Journal of Chemical Technology and Biotechnology
ISSN :
0268-2575
Publisher :
John Wiley & Sons Ltd., Chichester, United Kingdom
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
F.R.S.-FNRS - Fonds de la Recherche Scientifique ERA-NET eraIB (industrial biotechnology)
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