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Nat Protoc. 2019 May;14(5):1455-1488. doi: 10.1038/s41596-019-0146-6. Epub 2019 Apr 5.

Quantitative imaging of sleep behavior in Caenorhabditis elegans and larval Drosophila melanogaster.

Author information

1
Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, Philadelphia, PA, USA.
2
Department of Psychiatry, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
3
Department of Neurology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
4
Center for Excellence in Environmental Toxicology (CEET), Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
5
Chronobiology Program, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
6
Center for Sleep and Circadian Neurobiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
7
Department of Neuroscience, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
8
Department of Psychiatry, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. kayser@pennmedicine.upenn.edu.
9
Chronobiology Program, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. kayser@pennmedicine.upenn.edu.
10
Center for Sleep and Circadian Neurobiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. kayser@pennmedicine.upenn.edu.
11
Department of Neuroscience, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. kayser@pennmedicine.upenn.edu.

Abstract

Sleep is nearly universal among animals, yet remains poorly understood. Recent work has leveraged simple model organisms, such as Caenorhabditis elegans and Drosophila melanogaster larvae, to investigate the genetic and neural bases of sleep. However, manual methods of recording sleep behavior in these systems are labor intensive and low in throughput. To address these limitations, we developed methods for quantitative imaging of individual animals cultivated in custom microfabricated multiwell substrates, and used them to elucidate molecular mechanisms underlying sleep. Here, we describe the steps necessary to design, produce, and image these plates, as well as analyze the resulting behavioral data. We also describe approaches for experimentally manipulating sleep. Following these procedures, after ~2 h of experimental preparation, we are able to simultaneously image 24 C. elegans from the second larval stage to adult stages or 20 Drosophila larvae during the second instar life stage at a spatial resolution of 10 or 27 ┬Ám, respectively. Although this system has been optimized to measure activity and quiescence in Caenorhabditis larvae and adults and in Drosophila larvae, it can also be used to assess other behaviors over short or long periods. Moreover, with minor modifications, it can be adapted for the behavioral monitoring of a wide range of small animals.

PMID:
30953041
DOI:
10.1038/s41596-019-0146-6
[Indexed for MEDLINE]

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