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

William E. Pittman, Drew B. Sinha, William B. Zhang, Holly E. Kinser, Zachary Pincus

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

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.

Original languageEnglish
Pages (from-to)3909-3920
Number of pages12
JournalLab on a Chip
Volume17
Issue number22
DOIs
StatePublished - Nov 21 2017

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