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

ABSTRACT:

Background

Microbes live in dynamic environments where nutrient concentrations fluctuate. Quantifying fitness in terms of birth rate and death rate in a wide range of environments is critical for understanding microbial evolution and ecology.

Methods

Here, using high-throughput time-lapse microscopy, we have quantified how Saccharomyces cerevisiae mutants incapable of synthesizing an essential metabolite (auxotrophs) grow or die in various concentrations of the required metabolite. We establish that cells normally expressing fluorescent proteins lose fluorescence upon death and that the total fluorescence in an imaging frame is proportional to the number of live cells even when cells form multiple layers. We validate our microscopy approach of measuring birth and deat

SUBMITTER: Hart SFM 

PROVIDER: S-EPMC6785046 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature