A co-culture microplate platform to quantify microbial interactions and growth dynamics

Abstract

This thesis reports the development of BioMe, a co-culture microplate platform that enables high-throughput, real-time quantitative growth dynamics measurements of interacting microbial batch cultures. The primary BioMe components can be 3D-printed, allowing ease of fabrication and DIY accessibility in the microbiome community. A pairwise 3D-printed iteration of the BioMe device was used in diffusion and co-culture experiments. Genetically engineered Escherichia Coli lysine and isoleucine auxotroph strains were used to characterize the diffusion of amino acids across the porous membranes. Results demonstrated a nonlinear relationship between growth rate and pore size and also distinct diffusion behavior for lysine and isoleucine. Pairwise syntrophic co-culture experiments demonstrated synergistic but repressed interaction between these two paired auxotrophs. Investigation of the effect of varying initial amino acid conditions on growth dynamics demonstrated that small changes in initial media condition can consistently affect patterns of yield and growth rate of constituent microbial species.