PLoS ONE (Jan 2012)

A multi-platform flow device for microbial (co-) cultivation and microscopic analysis.

  • Matthijn C Hesselman,
  • Dorett I Odoni,
  • Brendan M Ryback,
  • Suzette de Groot,
  • Ruben G A van Heck,
  • Jaap Keijsers,
  • Pim Kolkman,
  • David Nieuwenhuijse,
  • Youri M van Nuland,
  • Erik Sebus,
  • Rob Spee,
  • Hugo de Vries,
  • Marten T Wapenaar,
  • Colin J Ingham,
  • Karin Schroën,
  • Vítor A P Martins dos Santos,
  • Sebastiaan K Spaans,
  • Floor Hugenholtz,
  • Mark W J van Passel

DOI
https://doi.org/10.1371/journal.pone.0036982
Journal volume & issue
Vol. 7, no. 5
p. e36982

Abstract

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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.