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

PLoS One. 2012;7(5):e36982. doi: 10.1371/journal.pone.0036982. Epub 2012 May 14.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Coculture Techniques / instrumentation
  • Equipment Design
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Genetic Engineering
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • High-Throughput Screening Assays / instrumentation
  • Microbiological Techniques / instrumentation*
  • Microfluidic Analytical Techniques / instrumentation*
  • Microscopy, Fluorescence
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism

Substances

  • Recombinant Proteins
  • Green Fluorescent Proteins