AUTHOR=Vuono David C. , Lipp Bruce , Staub Carl , Loney Evan , Harrold Zoë R. , Grzymski Joseph J. 

TITLE=A Real-Time Multiplexed Microbial Growth Intervalometer for Capturing High-Resolution Growth Curves

JOURNAL=Frontiers in Microbiology

VOLUME=Volume 10 - 2019

YEAR=2019

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2019.01135

DOI=10.3389/fmicb.2019.01135

ISSN=1664-302X

ABSTRACT=Batch cultures are a low maintenance and routine culturing method in microbiology. Automated tools that measure growth curves from microorganisms grown in traditional laboratory glassware, such as Balch-type tubes, are not commercially available. Here we present a new MicrobiAl Growth Intervalometer (MAGI) that measures optical density as it correlates to microbial growth by utilizing photo-conduction as opposed to photo-attenuation used by traditional OD measurement equipment. Photo-attenuation occurs when biomass in suspension within a medium, blocks and/or diffuses light directed at the detector, such that an increase in biomass results in a decrease in the measured signal. Photo-conduction differs in that the biomass contained in a medium conducts light from the emitter to the detector, where an increase in the biomass results in a corresponding increase in the measured signal. MAGI features software-driven automation that provides investigators with a highly sensitive, low background noise growth measurement instrument with added capabilities for remote visualization and data acquisition. It is a low maintenance, cost effective, versatile, and robust platform for aerobic/anaerobic cultivation. We demonstrate the versatility of this device by obtaining growth curves from two common laboratory organisms Escherichia coli K-12 and Bacillus subtilis. We show that growth rates and generation times in E. coli K-12 are reproducible to previously published results and that morphological changes of B. subtilis during growth can be detected as a change in the slope of the growth curve, which is a function of the effects of cell size on photo-conduction through the medium. We also test MAGI to capture growth curves from an environmental organism, Intrasporangium calvum C5, under various media compositions. Our results demonstrate that the MAGI platform accurately measures growth curves in media under various redox conditions (aerobic, microaerobic and anaerobic), complex and minimal medias, and resolving diauxic growth curves when I. calvum is grown on a disaccharide. Lastly, we demonstrate that the device can resolve growth curves for µM concentrations of resources that yield low biomass. This research advances the tools available to microbiologists aiming to monitor growth curves in a variety of disciplines, such as environmental microbiology, clinical microbiology, and food sciences.