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

Bio Protoc. 2018 Mar 20;8(6):e2774. doi: 10.21769/BioProtoc.2774.

Abstract

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 in vivo intracellular ROS levels photometrically in Caenorhabditis elegans using 2',7'-dichlorofluorescein diacetate (H2DCFDA), a cell permeant tracer.

Keywords: 2′,7′-Dichlorofluorescein diacetate; Antioxidant; C. elegans; Reactive oxygen species.