Method: Bacterial Lobster Traps Mask-based multi-photon lithography (MPL) is a versatile micro-fabrication approach developed in the Shear Group that has the ability to pattern porous hydrogel materials with sub-micron spatial resolution in three dimensions. In this direct-write fabrication method, solid structures are formed from aqueous protein precursors via covalent cross-linking reactions between amino acid residues in the focal volume of a pulsed laser beam. Complex three-dimensional microstructures can be formed layer-by-layer by translating the focus of the scanning beam in the optical (z) axis through solution. In collaboration with Prof. Jason Shear’s laboratory at UT Austin, we employ this highly adaptable fabrication platform to create customized microstructures to house small bacterial populations and use microscopy (light and confocal) to monitor group behaviors in real time.
Approach: Structures made using protein MPL have facilitated phenotypic studies of small aggregates of cells. A single bacterium can be captured within a picoliter-sized container. These porous bacterial “lobster traps,” facilitate normal cell growth, which can be observed using standard light and confocal microscopy techniques One cell enclosed in a three-dimensional user-defined geometry can become a clonal population quickly. We have studied growth kinetics, bacterial cell-cell communication (quorum sensing), and antibiotic resistance phenotypes of small, densely packed populations. Currently, we aim to characterize how sensing small molecules, such as nutrients (e.g., O2) and metabolic byproducts (e.g., H2O2) impact microbial behavior in high density, low cell number environments.