By What is DCFH-DA Assay?
Dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay: a quantitative method for oxidative stress assessment of nanoparticle-treated cells. Toxicol In Vitro.
What does DCFDA measure?
DCFDA / H2DCFDA / DCFH-DA / DCFH is a fluorogenic dye that measures hydroxyl, peroxyl and other reactive oxygen species (ROS) activity within the cell. It is then deacetylated by cellular esterases to a non-fluorescent compound, which is later oxidized by ROS into 2′, 7′ –dichlorofluorescein (DCF).
What is DCFH?
Dichlorofluorescin (DCFH) is a probe that is trapped within cells and is easily oxidized to fluorescent dichlorofluorescein (DCF). By itself, dichlorofluorescin (DCFH) also quantifies intracellular hydrogen peroxide as well as cellular oxidative stress.
How do you test for ROS?
Assess ROS by immediately analyzing your cells by flow cytometry using the FL1 channel (green fluorescence), or by fluorescence plate reader, or by fluorescence microscopy. The fluorescence can be detected by using excitation and emission wavelengths that are appropriate for green fluorescence.
How is intracellular ROS measured?
Another strength is the ability to localize cellular production of ROS within the tissue, and the sensitivity to detect ROS within a single cell. Analysis of intracellular H2O2 with DCFH-DA by flow cytometry or imaging has also been used to measure intracellular ROS.
What is ROS in cell?
Reactive oxygen species (ROS) serve as cell signaling molecules for normal biologic processes. However, the generation of ROS can also provoke damage to multiple cellular organelles and processes, which can ultimately disrupt normal physiology.
What is the difference between ROS and free radicals?
Free radicals are reactive chemicals with an unpaired electron in an outer orbit [1]. Reactive oxygen species (ROS) comprise both free radical and nonfree radical oxygen containing molecules such as hydrogen peroxide (H2O2), superoxide ( ), singlet oxygen (1/2O2), and the hydroxyl radical ( ).