Timm, C. M.Hansen, Ryan R.Doktycz, M. J.Retterer, S. T.Pelletier, D. A.2016-04-042016-04-042015-11-01http://hdl.handle.net/2097/32254Citation: Timm, C. M., Hansen, R. R., Doktycz, M. J., Retterer, S. T., & Pelletier, D. A. (2015). Microstencils to generate defined, multi-species patterns of bacteria. Biomicrofluidics, 9(6). doi:10.1063/1.4935938Microbial communities are complex heterogeneous systems that are influenced by physical and chemical interactions with their environment, host, and community members. Techniques that facilitate the quantitative evaluation of how microscale organization influences the morphogenesis of multispecies communities could provide valuable insights into the dynamic behavior and organization of natural communities, the design of synthetic environments for multispecies culture, and the engineering of artificial consortia. In this work, we demonstrate a method for patterning microbes into simple arrangements that allow the quantitative measurement of growth dynamics as a function of their proximity to one another. The method combines parylene-based liftoff techniques with microfluidic delivery to simultaneously pattern multiple bacterial species with high viability using low-cost, customizable methods. Quantitative measurements of bacterial growth for two competing isolates demonstrate that spatial coordination can play a critical role in multispecies growth and structure. © 2015 AIP Publishing LLC.This Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s).https://rightsstatements.org/page/InC/1.0/MicroorganismsChemical InteractionsComplex Heterogeneous SystemsMicrobial CommunitiesMicrofluidic DeliveryMulti-Species CommunitiesArticle