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Quantitative Assessment of Fat Levels in Caenorhabditis elegans Using Dark Field Microscopy

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...

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Autores principales: Fouad, Anthony D., Pu, Shelley H., Teng, Shelly, Mark, Julian R., Fu, Moyu, Zhang, Kevin, Huang, Jonathan, Raizen, David M., Fang-Yen, Christopher
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Genetics Society of America 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5473760/
https://www.ncbi.nlm.nih.gov/pubmed/28404661
http://dx.doi.org/10.1534/g3.117.040840
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author Fouad, Anthony D.
Pu, Shelley H.
Teng, Shelly
Mark, Julian R.
Fu, Moyu
Zhang, Kevin
Huang, Jonathan
Raizen, David M.
Fang-Yen, Christopher
author_facet Fouad, Anthony D.
Pu, Shelley H.
Teng, Shelly
Mark, Julian R.
Fu, Moyu
Zhang, Kevin
Huang, Jonathan
Raizen, David M.
Fang-Yen, Christopher
author_sort Fouad, Anthony D.
collection PubMed
description 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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spelling pubmed-54737602017-06-27 Quantitative Assessment of Fat Levels in Caenorhabditis elegans Using Dark Field Microscopy Fouad, Anthony D. Pu, Shelley H. Teng, Shelly Mark, Julian R. Fu, Moyu Zhang, Kevin Huang, Jonathan Raizen, David M. Fang-Yen, Christopher G3 (Bethesda) Investigations 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. Genetics Society of America 2017-04-12 /pmc/articles/PMC5473760/ /pubmed/28404661 http://dx.doi.org/10.1534/g3.117.040840 Text en Copyright © 2017 Fouad et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Investigations
Fouad, Anthony D.
Pu, Shelley H.
Teng, Shelly
Mark, Julian R.
Fu, Moyu
Zhang, Kevin
Huang, Jonathan
Raizen, David M.
Fang-Yen, Christopher
Quantitative Assessment of Fat Levels in Caenorhabditis elegans Using Dark Field Microscopy
title Quantitative Assessment of Fat Levels in Caenorhabditis elegans Using Dark Field Microscopy
title_full Quantitative Assessment of Fat Levels in Caenorhabditis elegans Using Dark Field Microscopy
title_fullStr Quantitative Assessment of Fat Levels in Caenorhabditis elegans Using Dark Field Microscopy
title_full_unstemmed Quantitative Assessment of Fat Levels in Caenorhabditis elegans Using Dark Field Microscopy
title_short Quantitative Assessment of Fat Levels in Caenorhabditis elegans Using Dark Field Microscopy
title_sort quantitative assessment of fat levels in caenorhabditis elegans using dark field microscopy
topic Investigations
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5473760/
https://www.ncbi.nlm.nih.gov/pubmed/28404661
http://dx.doi.org/10.1534/g3.117.040840
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