Development of a microscale aqueous two-phase system bacterial culture platform

Development of a bacterial culture platform which accounts for the characteristic human gut oxygen gradient could allow for improved understanding of the gut microbiome and its implications in human health as well as targeted therapies for related diseases. This work presents a microscale bacterial culture platform in an aqueous two-phase system (ATPS) with integrated oxygen-sensing microbeads. Generation of the oxygen gradient is explored through the culture of the facultative bacterium, Escherichia coli (E. coli), within the platform. Growth characterization of GFP-expressing E.coli O157:H7 within the platform is achieved through time lapse measurements of oxygen consumption and GFP fluorescence, as well as plate counting data. Phase fluorimetry measurements are collected through the oxygen-sensing microbeads placed at the bottom of the culture wells. Results indicate that E.coli was able to grow in the ATPS culture environment and that near anaerobic conditions were achieved at the bottom of the culture wells. Oxygen concentration information is restricted to the bottom of the culture wells due to microbead placement; however, in future iterations, this platform could accommodate other types of bacteria such as aerobic and anaerobic bacteria.Undergraduat