title: A multi-platform flow device for microbial (co-) cultivation and microscopic analysis.
abstract: Novel microbial cultivation platforms are of increasing interest to researchers in academia and industry. The development of materials with specialized chemical and geometric properties has opened up new possibilities in the study of previously unculturable microorganisms and has facilitated the design of elegant, high-throughput experimental set-ups. Within the context of the international Genetically Engineered Machine (iGEM) competition, we set out to design, manufacture, and implement a flow device that can accommodate multiple growth platforms, that is, a silicon nitride based microsieve and a porous aluminium oxide based microdish. It provides control over (co-)culturing conditions similar to a chemostat, while allowing organisms to be observed microscopically. The device was designed to be affordable, reusable, and above all, versatile. To test its functionality and general utility, we performed multiple experiments with Escherichia coli cells harboring synthetic gene circuits and were able to quantitatively study emerging expression dynamics in real-time via fluorescence microscopy. Furthermore, we demonstrated that the device provides a unique environment for the cultivation of nematodes, suggesting that the device could also prove useful in microscopy studies of multicellular microorganisms.
authors.affiliation: Wageningen UR iGEM 2011 Team, Wageningen University, Wageningen, The Netherlands.
authors.forename: Matthijn C
authors.initials: MC
authors.lastname: Hesselman
authors.forename: Dorett I
authors.initials: DI
authors.lastname: Odoni
authors.forename: Brendan M
authors.initials: BM
authors.lastname: Ryback
authors.forename: Suzette
authors.initials: S
authors.lastname: de Groot
authors.forename: Ruben G A
authors.initials: RG
authors.lastname: van Heck
authors.forename: Jaap
authors.initials: J
authors.lastname: Keijsers
authors.forename: Pim
authors.initials: P
authors.lastname: Kolkman
authors.forename: David
authors.initials: D
authors.lastname: Nieuwenhuijse
authors.forename: Youri M
authors.initials: YM
authors.lastname: van Nuland
authors.forename: Erik
authors.initials: E
authors.lastname: Sebus
authors.forename: Rob
authors.initials: R
authors.lastname: Spee
authors.forename: Hugo
authors.initials: H
authors.lastname: de Vries
authors.forename: Marten T
authors.initials: MT
authors.lastname: Wapenaar
authors.forename: Colin J
authors.initials: CJ
authors.lastname: Ingham
authors.forename: Karin
authors.initials: K
authors.lastname: Schroën
authors.forename: Vítor A P
authors.initials: VA
authors.lastname: Martins dos Santos
authors.forename: Sebastiaan K
authors.initials: SK
authors.lastname: Spaans
authors.forename: Floor
authors.initials: F
authors.lastname: Hugenholtz
authors.forename: Mark W J
authors.initials: MW
authors.lastname: van Passel
mesh_terms: D018920:Coculture Techniques / Q000295:instrumentation; D004867:Equipment Design; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005818:Genetic Engineering; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000295:instrumentation; D008828:Microbiological Techniques / Q000295:instrumentation*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D008856:Microscopy, Fluorescence; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins