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Issue 22, 2017
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A simple culture system for long-term imaging of individual C. elegans

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Abstract

We have miniaturized standard culture techniques to rear arrays of isolated, individual C. elegans throughout their lives on solid gel media. The resulting apparatus is compatible with brightfield and fluorescence microscopy, enabling longitudinal studies of morphology and fluorescent transgene expression. Our culture system exploits a novel crosslinking reaction between a polyethylene glycol hydrogel and a silicone elastomer to constrain animals to individual “corrals” on the gel surface. These devices are simple to construct on the benchtop with commercially available reagents, and, unlike microfluidic isolation methods, do not rely on micropatterned materials. We demonstrate that this new culture method has negligible effects on the physiology of C. elegans compared to standard culture on agar plates. In addition, RNAi techniques are effective in this system. Finally, the hydrogel–silicone binding chemistry that we developed also allows traditional microfluidic devices to be covalently attached to gel substrates instead of glass.

Graphical abstract: A simple culture system for long-term imaging of individual C. elegans

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Publication details

The article was received on 25 Aug 2017, accepted on 15 Oct 2017 and first published on 16 Oct 2017


Article type: Paper
DOI: 10.1039/C7LC00916J
Citation: Lab Chip, 2017,17, 3909-3920
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    A simple culture system for long-term imaging of individual C. elegans

    W. E. Pittman, D. B. Sinha, W. B. Zhang, H. E. Kinser and Z. Pincus, Lab Chip, 2017, 17, 3909
    DOI: 10.1039/C7LC00916J

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