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High-throughput quantification of microbial birth and death dynamics using fluorescence microscopy.

Samuel F M Hart¹, David Skelding¹, Adam J Waite¹

¹Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.

Quantitative Biology (Beijing, China)

|October 11, 2019

Summary

Quantifying microbial birth and death rates is crucial for understanding evolution. This study uses time-lapse microscopy to reveal non-intuitive microbial dynamics in fluctuating nutrient environments.

Keywords:

Saccharomyces cerevisiae birth rate death rate fluorescence microscopy microbial growth

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Area of Science:

Microbial Ecology
Evolutionary Biology
Cell Biology

Background:

Microbial environments feature fluctuating nutrient concentrations.
Quantifying microbial birth and death rates is essential for understanding microbial evolution and ecology.

Purpose of the Study:

To quantify the growth and death dynamics of Saccharomyces cerevisiae auxotrophs across various essential metabolite concentrations.
To establish a robust method for measuring microbial birth and death rates using time-lapse microscopy.

Main Methods:

High-throughput time-lapse microscopy was employed to monitor auxotrophic yeast mutants.
Fluorescence loss upon cell death was used as a viability indicator.
Microscopy-based birth and death rate measurements were validated using flow cytometry, cell counting, and chemostat culturing.

Main Results:

Low lysine concentrations delayed death but did not support birth in lysine-requiring cells.
Hypoxanthine-requiring cells exhibited increased death rates in low hypoxanthine, despite some cell production.
The Moser model better described birth rates than the Monod model across varying metabolite concentrations.

Conclusions:

Time-lapse microscopy effectively reveals complex microbial birth and death dynamics.
This approach enables precise quantification of growth rates in diverse environmental conditions.