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JoVE Journal
Biology
Quantification of Lipid Abundance and Evaluation of Lipid Distribution in Caenorhabditis eleg...
Quantification of Lipid Abundance and Evaluation of Lipid Distribution in Caenorhabditis eleg...
JoVE Journal
Biology
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JoVE Journal Biology
Quantification of Lipid Abundance and Evaluation of Lipid Distribution in Caenorhabditis elegans by Nile Red and Oil Red O Staining

Quantification of Lipid Abundance and Evaluation of Lipid Distribution in Caenorhabditis elegans by Nile Red and Oil Red O Staining

Full Text
21,586 Views
07:46 min
March 5, 2018

DOI: 10.3791/57352-v

Wilber Escorcia1, Dana L. Ruter1,2,3, James Nhan1,2, Sean P. Curran1,2

1Leonard Davis School of Gerontology,University of Southern California, 2Molecular & Computational Biology Section,University of Southern California, 3Integrative Program for Biological & Genome Sciences,University of North Carolina at Chapel Hill

Nile red staining of fixed Caenorhabditis elegans is a method for quantitative measurement of neutral lipid deposits, while oil red O staining facilitates qualitative assessment of lipid distribution among tissues.

The overall goal of these staining methods is to quantify intracellular lipid abundance and evaluate lipid distribution among different tissues. This method can help answer key questions in lipid metabolism such as how are lipids genetically and biochemically regulated and which disease states this regulate lipid homeostasis. The main advantage of this technique is that it is relatively simple and inexpensive to carry out.

To carry out Nile Red lipid staining, plate worms on Nematode Growth Medium or NGM seeded with late log OP50 E.coli and grow the worms at 20 degrees Celsius to early L4 stage. Wash the worms with one millimeter of PBST solution and transfer the worm suspension to a 1.5 milliliter microfuge tube. Centrifuge the worms at 560 times gravity for one minute.

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