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Multi-parameter flow cytometry and cell sorting reveal extensive physiological heterogeneity in Bacillus cereus batch
Andrew Want1, Helen Hancocks, Colin R Thomas
1Centre for Biological Engineering, Department of Chemical Engineering, Loughborough University, Leicestershire, UK.
Biotechnology Letters
|March 3, 2011
Summary
Bacillus cereus cultures exhibit complex population dynamics, with individual cells showing distinct physiological states and varying growth behaviors. This heterogeneity highlights how single bacterial cells respond differently to their environment.
Area of Science:
- Microbiology
- Cell Biology
- Biophysics
Background:
- Bacterial monocultures are often assumed to be homogenous.
- Understanding population dynamics is crucial for microbiology and biotechnology.
- Physiological heterogeneity can impact bacterial responses to stimuli.
Purpose of the Study:
- To investigate the physiological heterogeneity of Bacillus cereus during batch culture.
- To characterize distinct physiological states within a bacterial population.
- To analyze the colony formation and growth dynamics of individual Bacillus cereus cells.
Main Methods:
- Utilized multi-parameter flow cytometry and cell sorting.
- Employed two staining protocols: DiOC(6)(3)/propidium iodide (PI) and RedoxSensor Green/PI.
- Sorted individual cells onto nutrient agar plates for colony analysis.
Main Results:
- Identified at least four distinguishable physiological states in Bacillus cereus batch cultures.
- Observed significant variation in colony formation and lag phases among single sorted cells.
- Demonstrated that cells from the same monoculture exhibit markedly dissimilar growth behaviors.
Conclusions:
- Bacterial monocultures are physiologically heterogeneous and dynamic.
- Individual bacterial cells respond disparately to environmental conditions.
- The observed heterogeneity influences population dynamics and cellular responses.
