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Multi-parameter flow cytometry and cell sorting reveal extensive physiological heterogeneity in Bacillus cereus batch cultures

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Abstract

Based on two staining protocols, DiOC6(3)/propidium iodide (PI) and RedoxSensor Green (an indicator of bacterial reductase activity)/PI, multi-parameter flow cytometry and cell sorting has identified at least four distinguishable physiological states during batch cultures of Bacillus cereus. Furthermore, dependent on the position in the growth curve, single cells gave rise to varying numbers of colonies when sorted individually onto nutrient agar plates. These growing colonies derived from a single cell had widely different lag phases, inferred from differences in colony size. This further highlights the complex population dynamics of bacterial monocultures and further demonstrates that individual bacterial cells in a culture respond in markedly dissimilar ways to the environment, resulting in a physiologically heterogenous and dynamic population.

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Abbreviations

Ψ:

Electro potential

i:

Inside

o:

Outside

a :

Active (diffusible or unbound) concentration

R :

Universal gas constant

T :

Absolute temperature

Z :

Charge

F :

Faraday constant

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Acknowledgments

Fluorochromes were provided by Molecular Probes/Invitrogen (Oregon, USA) and the authors thank William Godfrey (Invitrogen Corporation) for his helpful discussions. The authors would like to acknowledge the financial support of the Biotechnology and Biological Sciences Research Council (UK), Avecia Biologics Ltd (UK) and Novozymes A/S (Denmark) for this study.

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Authors and Affiliations

  1. Centre for Biological Engineering, Department of Chemical Engineering, Loughborough University, Leicestershire, LE11 3TU, UK

    Andrew Want & Christopher J. Hewitt

  2. Centre for Bioprocess Engineering, School of Chemical Engineering, University of Birmingham, Edgbaston, B15 2TT, UK

    Helen Hancocks & Colin R. Thomas

  3. Novozymes A/S, Smørmosevej 25 (2JS.12), 2880, Bagsværd, Denmark

    Stuart M. Stocks

  4. Inverness Medical International, Priory Business Park, Bedford, MK44 3UP, UK

    Gerhard Nebe-von-Caron

Authors
  1. Andrew Want
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  2. Helen Hancocks
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  3. Colin R. Thomas
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  4. Stuart M. Stocks
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  5. Gerhard Nebe-von-Caron
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  6. Christopher J. Hewitt
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Correspondence to Christopher J. Hewitt.

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Want, A., Hancocks, H., Thomas, C.R. et al. Multi-parameter flow cytometry and cell sorting reveal extensive physiological heterogeneity in Bacillus cereus batch cultures. Biotechnol Lett 33, 1395–1405 (2011). https://doi.org/10.1007/s10529-011-0566-z

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  • DOI: https://doi.org/10.1007/s10529-011-0566-z

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