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

Hesselman MC, Odoni DI, Ryback BM, de Groot S, van Heck RG, Keijsers J, Kolkman P, Nieuwenhuijse D, van Nuland YM, Sebus E, Spee R, de Vries H, Wapenaar MT, Ingham CJ, Schroën K, Martins dos Santos VA, Spaans SK, Hugenholtz F, van Passel MW - PLoS ONE (2012)

Bottom Line: 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.The device was designed to be affordable, reusable, and above all, versatile.

View Article: PubMed Central - PubMed

Affiliation: Wageningen UR iGEM 2011 Team, Wageningen University, Wageningen, The Netherlands.

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.

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Cell retention on the microsieve.Retained clusters of fluorescent E. coli cells on the sieve (100x magnification). The diagonal light grey bands are the permeable areas of the microsieve that contain the pores.
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pone-0036982-g002: Cell retention on the microsieve.Retained clusters of fluorescent E. coli cells on the sieve (100x magnification). The diagonal light grey bands are the permeable areas of the microsieve that contain the pores.

Mentions: The device containing a microsieve was inoculated with fluorescent E. coli cells. The bacteria were retained predominantly on the diagonal permeable areas, where small clusters of cells could be discerned. Even after applying a gentle flow over the retained cells, potentially allowing exposure of the cells to other compounds in the context of biosensing, the cells remained trapped on the sieve (Figure 2).


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

Hesselman MC, Odoni DI, Ryback BM, de Groot S, van Heck RG, Keijsers J, Kolkman P, Nieuwenhuijse D, van Nuland YM, Sebus E, Spee R, de Vries H, Wapenaar MT, Ingham CJ, Schroën K, Martins dos Santos VA, Spaans SK, Hugenholtz F, van Passel MW - PLoS ONE (2012)

Cell retention on the microsieve.Retained clusters of fluorescent E. coli cells on the sieve (100x magnification). The diagonal light grey bands are the permeable areas of the microsieve that contain the pores.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3351485&req=5

pone-0036982-g002: Cell retention on the microsieve.Retained clusters of fluorescent E. coli cells on the sieve (100x magnification). The diagonal light grey bands are the permeable areas of the microsieve that contain the pores.
Mentions: The device containing a microsieve was inoculated with fluorescent E. coli cells. The bacteria were retained predominantly on the diagonal permeable areas, where small clusters of cells could be discerned. Even after applying a gentle flow over the retained cells, potentially allowing exposure of the cells to other compounds in the context of biosensing, the cells remained trapped on the sieve (Figure 2).

Bottom Line: 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.The device was designed to be affordable, reusable, and above all, versatile.

View Article: PubMed Central - PubMed

Affiliation: Wageningen UR iGEM 2011 Team, Wageningen University, Wageningen, The Netherlands.

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.

Show MeSH
Related in: MedlinePlus