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. 2021 Aug 20;11(16):e4124.
doi: 10.21769/BioProtoc.4124.

Oil Red O Staining for Lipid Content in Caenorhabditis elegans

Feng-Yung Wang  1 , Tsui-Ting Ching  2
Affiliations

Oil Red O Staining for Lipid Content in Caenorhabditis elegans

Feng-Yung Wang et al. Bio Protoc. .

Abstract

The nematode Caenorhabditis elegans has emerged as a popular model system for studying the regulation of lipid metabolism. Therefore, it is critical to develop a method for determining fat storage in individual worms. Oil Red O (ORO) staining has been validated as an accurate assessment for major fat storage in C. elegans. Here, we describe an optimized protocol for ORO staining of C. elegans and provide detailed instructions for quantifying the intensity of ORO signal in images acquired by light microscopy.

Keywords: C. elegans; Fat; Lipid metabolism; Oil Red O.

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Conflict of interest statement

Competing interestsThe authors have no conflicts of interest to declare.

Figures

Figure 1.
Figure 1.. User interface of the ORO staining software.
A. The “Image” window. The intensity values of pixels in the red area will be calculated as ORO signals. B. The “Staining quantify” window includes two threshold selection bars, the result chart, and the buttons for settings. C. The “Files” window. The images opened will be listed in this window.
Figure 2.
Figure 2.. Oil Red O staining levels in wild-type N2, eat-2(ad1116), and daf-2(e1370) mutants.
A. Representative images of Oil Red O staining in wild-type N2, eat-2(ad1116), and daf-2(e1370) animals at larval 4 stage (Scale bar: 50 μm). B. Quantification of the area stained by the Oil Red O in wild-type N2, eat-2(ad1116), and daf-2(e1370) mutants. C. Quantification of the average Oil Red O intensity in wild-type animals, eat-2(ad1116), and daf-2(e1370) mutants (100% describes the level of saturation). One-way ANOVA test followed by Dunnett's post-hoc test for selected groups was used. Sample size: n = 40 for N2, n = 45 for eat-2, n = 30 for daf-2. Results represent mean ± SD, and *** P < 0.001.
See this image and copyright information in PMC

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