Investigating sociomicrobiology by integrating micro 3D printing with quantitative analytical techniques

Antibiotic resistant polymicrobial infections have become a source of great concern in recent years both in clinical settings as well as in basic and medical research. Incidence of resistance and increased virulence, which typically emerge within small, dense cellular ensembles on picoliter scales, is on the rise and scientists are just beginning to understand the complexity of these dangerous bacterial populations. To that end, the research in this dissertation has sought to analyze the complex social interactions of micro 3D (µ3D) printed bacterial colonies with a variety of analytical techniques. Through characterization of the µ3D printed hydrogels themselves, and by pairing this technology with fluorescence and confocal microscopy, electrochemical studies, and mass spectrometry, important insights regarding the sociomicrobiology of these bacterial communities emerge. The Shear lab has previously employed µ3D printing of bacterial aggregates to study microbial populations in environments that reproduce attributes associated with complex spatiotemporal in vivo conditions to a much greater extent than traditional culture techniques. Combining this technology with advanced imaging approaches has enabled a detailed investigation into properties of intra- and inter-species cooperation, including factors that influence antibiotic resistance and virulence. The goal of the work presented here is to integrate quantitative and qualitative analytical techniques with µ3D printing technology to enable novel approaches for studying interactions, both within and between small bacterial aggregates in complex microbial environments. This information will be vital in the next steps toward designing better and more efficient strategies for combating complex pathogenic communities that exist within polymicrobial infection environments.