Measurement of Intracellular ROS in Caenorhabditis elegans Using 2',7'-Dichlorodihydrofluorescein Diacetate

Reactive oxygen species (ROS) are generated during normal metabolic processes under aerobic conditions. Since ROS production initiates harmful radical chain reactions on cellular macromolecules, including lipid peroxidation, DNA mutation, and protein denaturation, it has been implicated in a wide sp...

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Bibliographic Details
Published inBio-protocol Vol. 8; no. 6
Main Authors Yoon, Dong Suk, Lee, Myon-Hee, Cha, Dong Seok
Format Journal Article
LanguageEnglish
Published United States Bio-Protocol 20.03.2018
Bio-protocol LLC
Subjects
Biology
Methods
C. elegans
Reactive oxygen species
2′,7′-Dichlorofluorescein diacetate
Antioxidant
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Summary:Reactive oxygen species (ROS) are generated during normal metabolic processes under aerobic conditions. Since ROS production initiates harmful radical chain reactions on cellular macromolecules, including lipid peroxidation, DNA mutation, and protein denaturation, it has been implicated in a wide spectrum of diseases such as cancer, cardiovascular disease, ischemia-reperfusion and aging. Over the past several decades, antioxidants have received explosive attention regarding their protective potential against these deleterious reactions. Accordingly, many analytical methodologies have been developed for the evaluation of the antioxidant capacity of compounds or complex biological samples. Herein, we introduce a simple and convenient method to detect intracellular ROS levels photometrically in using 2',7'-dichlorofluorescein diacetate (H DCFDA), a cell permeant tracer.
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ISSN:2331-8325
2331-8325
DOI:10.21769/bioprotoc.2774