Projection Microstereolithographic Microbial Bioprinting for Engineered Biofilms

Microbes are critical drivers of all ecosystems and many biogeochemical processes, yet little is known about how the three-dimensional (3D) organization of these dynamic organisms contributes to their overall function. To probe how biofilm structure affects microbial activity, we developed a technique for patterning microbes in 3D geometries using projection stereolithography to bioprint microbes within hydrogel architectures. Bacteria were printed and monitored for biomass accumulation, demonstrating postprint viability of cells using this technique. We verified our ability to integrate biological and geometric complexity by fabricating a printed biofilm with two E. coli strains expressing different fluorescence. Finally, we examined the target application of microbial absorption of metal ions to investigate geometric effects on both the metal sequestration efficiency and the uranium sensing capability of patterned engineered Caulobacter crescentus strains. This work represents the first demonstration of the stereolithographic printing of microbials and presents opportunities for future work of engineered biofilms and other complex 3D structured cultures.

Automated summary

In this study, a technique for patterning microbes in 3D geometries using projection stereolithography was developed and successfully used to demonstrate the impact of biofilm structure on microbial activity. The ability to print bacteria and fabricate a biofilm with different fluorescence E. coli strains was verified, showing potential for future work on engineered biofilms and complex 3D structures in microbial cultures.