Imaging the oxygen wave with a single bioluminescent bacterium

Abstract

We developed a capability of a monolayer of bioluminescent (BL) bacteria for spatiotemporally visualizing the heterogeneous distribution and dynamic evolution of interfacial oxygen concentration, resulting in the discovery of spontaneous and stochastic oxygen waves at the interface between the substrate and an undisturbed, apparently still solution. Wild type bacteria, P. phosphoreum, spontaneously emit light during the native metabolism processes, i.e., bioluminescence. The emission intensity is sensitively regulated by oxygen concentration. By taking the electrolysis of water as a model, it was demonstrated that time-lapsed BL imaging of a bacterial monolayer allowed for visualizing the dynamic distribution of oxygen. The results were quantitatively understood with a physical model involving the diffusion equation and Michaelis–Menten equation. Unexpectedly, further study uncovered a spontaneous and stochastic oxygen wave in a standard well of a microtiter plate, which was subsequently attributed to the inevitable micro-convections induced by inhomogeneous evaporation and thermal fluctuation. Because of the wide application of microtiter plates, this study sheds new light to better understand the apparent heterogeneity in cell-culture and bio-assays.