A Multi-Platform Flow Device for Microbial (Co-) Cultivation and Microscopic Analysis

M.C. Hesselman, D.I. Odoni, B.M. Ryback, S. de Groot, R.G.A. van Heck, J. Keijsers, P. Kolkman, D. Nieuwenhuijse, Y.M. Nuland, E. Sebus, R. Spee, H. de Vries, M.T. Wapenaar, C.J. Ingham, K. Schroen, V.A.P. Martins Dos Santos, S.K. Spaans, F. Hugenholtz, M.W.J. van Passel

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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
Original languageEnglish
Article numbere36982
JournalPLoS ONE
Volume7
Issue number5
DOIs
Publication statusPublished - 2012

Keywords

  • culture
  • microorganisms
  • population
  • resistance
  • membranes
  • bacteria
  • chamber
  • support
  • arrays

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    Hesselman, M. C., Odoni, D. I., Ryback, B. M., de Groot, S., van Heck, R. G. A., Keijsers, J., Kolkman, P., Nieuwenhuijse, D., Nuland, Y. M., Sebus, E., Spee, R., de Vries, H., Wapenaar, M. T., Ingham, C. J., Schroen, K., Martins Dos Santos, V. A. P., Spaans, S. K., Hugenholtz, F., & van Passel, M. W. J. (2012). A Multi-Platform Flow Device for Microbial (Co-) Cultivation and Microscopic Analysis. PLoS ONE, 7(5), [e36982]. https://doi.org/10.1371/journal.pone.0036982