Quantitative assessment of fat levels in Caenorhabditis elegans using dark field microscopy

Anthony D. Fouad; Shelley H. Pu; Shelly Teng; Julian R. Mark; Moyu Fu; Kevin Zhang; Jonathan Huang; David M. Raizen; Christopher Fang-Yen

doi:10.1534/g3.117.040840

Quantitative assessment of fat levels in Caenorhabditis elegans using dark field microscopy

Anthony D. Fouad, Shelley H. Pu, Shelly Teng, Julian R. Mark, Moyu Fu, Kevin Zhang, Jonathan Huang, David M. Raizen, Christopher Fang-Yen

College of Science

Research output: Contribution to journal › Article › peer-review

12 Citations (Scopus)

Abstract

The roundworm Caenorhabditis elegans is widely used as a model for studying conserved pathways for fat storage, aging, and metabolism. The most broadly used methods for imaging fat in C. elegans require fixing and staining the animal. Here, we show that dark field images acquired through an ordinary light microscope can be used to estimate fat levels in worms. We define a metric based on the amount of light scattered per area, and show that this light scattering metric is strongly correlated with worm fat levels as measured by Oil Red O (ORO) staining across a wide variety of genetic backgrounds and feeding conditions. Dark field imaging requires no exogenous agents or chemical fixation, making it compatible with live worm imaging. Using our method, we track fat storage with high temporal resolution in developing larvae, and show that fat storage in the intestine increases in at least one burst during development.

Original language	English
Pages (from-to)	1811-1818
Number of pages	8
Journal	G3: Genes, Genomes, Genetics
Volume	7
Issue number	6
DOIs	https://doi.org/10.1534/g3.117.040840
Publication status	Published - 2017 Jun 1

Bibliographical note

Funding Information:
We thank Jianting Wang and Yu Chen of the University of Maryland, College Park, for providing the scattering phantom raw material. Some strains were provided by the CGC, which is funded by the National Institutes of Health (NIH) Office of Research Infrastructure Programs (P40 OD010440). A.D.F. (5R01NS084835-03) and D.M.R. (R01NS088432 and R21NS091500) were supported by the NIH. C.F.-Y. was supported by the NIH (5R01NS084835-03), the Ellison Medical Foundation, and the Alfred P. Sloan Research Foundation

Publisher Copyright:
© 2017 Fouad et al.

Keywords

C. elegans
Dark field fat
Nile Red
Oil Red O

ASJC Scopus subject areas

Molecular Biology
Genetics
Genetics(clinical)

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10.1534/g3.117.040840

Cite this

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title = "Quantitative assessment of fat levels in Caenorhabditis elegans using dark field microscopy",

abstract = "The roundworm Caenorhabditis elegans is widely used as a model for studying conserved pathways for fat storage, aging, and metabolism. The most broadly used methods for imaging fat in C. elegans require fixing and staining the animal. Here, we show that dark field images acquired through an ordinary light microscope can be used to estimate fat levels in worms. We define a metric based on the amount of light scattered per area, and show that this light scattering metric is strongly correlated with worm fat levels as measured by Oil Red O (ORO) staining across a wide variety of genetic backgrounds and feeding conditions. Dark field imaging requires no exogenous agents or chemical fixation, making it compatible with live worm imaging. Using our method, we track fat storage with high temporal resolution in developing larvae, and show that fat storage in the intestine increases in at least one burst during development.",

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note = "Funding Information: We thank Jianting Wang and Yu Chen of the University of Maryland, College Park, for providing the scattering phantom raw material. Some strains were provided by the CGC, which is funded by the National Institutes of Health (NIH) Office of Research Infrastructure Programs (P40 OD010440). A.D.F. (5R01NS084835-03) and D.M.R. (R01NS088432 and R21NS091500) were supported by the NIH. C.F.-Y. was supported by the NIH (5R01NS084835-03), the Ellison Medical Foundation, and the Alfred P. Sloan Research Foundation Publisher Copyright: {\textcopyright} 2017 Fouad et al.",

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AU - Huang, Jonathan

AU - Raizen, David M.

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N2 - The roundworm Caenorhabditis elegans is widely used as a model for studying conserved pathways for fat storage, aging, and metabolism. The most broadly used methods for imaging fat in C. elegans require fixing and staining the animal. Here, we show that dark field images acquired through an ordinary light microscope can be used to estimate fat levels in worms. We define a metric based on the amount of light scattered per area, and show that this light scattering metric is strongly correlated with worm fat levels as measured by Oil Red O (ORO) staining across a wide variety of genetic backgrounds and feeding conditions. Dark field imaging requires no exogenous agents or chemical fixation, making it compatible with live worm imaging. Using our method, we track fat storage with high temporal resolution in developing larvae, and show that fat storage in the intestine increases in at least one burst during development.

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Quantitative assessment of fat levels in Caenorhabditis elegans using dark field microscopy

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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