Dataset Information

Multi-well imaging of development and behavior in Caenorhabditis elegans.

ABSTRACT: The nematode Caenorhabditis elegans is widely used as a model for understanding the neuronal and genetic bases of behavior. Recent studies have required longitudinal assessment of individual animal's behavior over extended periods.Here we present a technique for automated monitoring of multiple worms for several days. Our method uses an array of plano-concave glass wells containing standard agar media. The concave well geometry allows worms to be imaged even at the edge of the agar surface and prevents them from burrowing under the agar. We transfer one worm or embryo into each well, and perform imaging of the array of wells using a single camera. Machine vision software is used to quantify size, activity, and/or fluorescence of each worm over time.We demonstrate the utility of our method in two applications: (1) quantifying behavioral quiescence and developmental rate in wild-type and mutant animals, and (2) characterizing differences in mating behavior between two C. elegans strains.Current techniques for tracking behavior in identified worms are generally restricted to imaging either single animals or have not been shown to work with arbitrary developmental stages; many are also technically complex. Our system works with up to 24 animals of any stages and is technically simple.Our multi-well imaging method is a powerful tool for quantification of long-term behavioral phenotypes in C. elegans.

SUBMITTER: Yu CC

PROVIDER: S-EPMC3972622 | biostudies-literature | 2014 Feb

REPOSITORIES: biostudies-literature

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Publications

Multi-well imaging of development and behavior in Caenorhabditis elegans.

Yu Chih-Chieh Jay CC Raizen David M DM Fang-Yen Christopher C

Journal of neuroscience methods 20131207

<h4>Background</h4>The nematode Caenorhabditis elegans is widely used as a model for understanding the neuronal and genetic bases of behavior. Recent studies have required longitudinal assessment of individual animal's behavior over extended periods.<h4>New method</h4>Here we present a technique for automated monitoring of multiple worms for several days. Our method uses an array of plano-concave glass wells containing standard agar media. The concave well geometry allows worms to be imaged even ...[more]

PMID: 24321627

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Project description:The nematode Caenorhabditis elegans is among the most common model systems used in aging research owing to its simple and inexpensive culture techniques, rapid reproduction cycle (~3 days), short lifespan (~3 weeks), and numerous available tools for genetic manipulation and molecular analysis. The most common approach for conducting aging studies in C. elegans, including survival analysis, involves culturing populations of tens to hundreds of animals together on solid nematode growth media (NGM) in Petri plates. While this approach gathers data on a population of animals, most protocols do not track individual animals over time. Presented here is an optimized protocol for the long-term culturing of individual animals on microfabricated polydimethylsiloxane (PDMS) devices called WorMotels. Each device allows up to 240 animals to be cultured in small wells containing NGM, with each well isolated by a copper sulfate-containing moat that prevents the animals from fleeing. Building on the original WorMotel description, this paper provides a detailed protocol for molding, preparing, and populating each device, with descriptions of common technical complications and advice for troubleshooting. Within this protocol are techniques for the consistent loading of small-volume NGM, the consistent drying of both the NGM and bacterial food, options for delivering pharmacological interventions, instructions for and practical limitations to reusing PDMS devices, and tips for minimizing desiccation, even in low-humidity environments. This technique allows the longitudinal monitoring of various physiological parameters, including stimulated activity, unstimulated activity, body size, movement geometry, healthspan, and survival, in an environment similar to the standard technique for group culture on solid media in Petri plates. This method is compatible with high-throughput data collection when used in conjunction with automated microscopy and analysis software. Finally, the limitations of this technique are discussed, as well as a comparison of this approach to a recently developed method that uses microtrays to culture isolated nematodes on solid media.

Dataset Information

Multi-well imaging of development and behavior in Caenorhabditis elegans.

Publications

Multi-well imaging of development and behavior in Caenorhabditis elegans.

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