Dichlorofluorescein (DCFH) oxidation assay measures hydrogen peroxide (H2O2), which is a derivative of superoxide anion. We found that a calmodulin antagonist, W-13, which is known to inhibit superoxide anion generation enhanced the capacity of human neutrophils to oxidize DCFH. To investigate this discrepancy we studied the role of nitric oxide (NO) in DCFH oxidation. Pure NO was capable of ox...

The use of antioxidants to prevent intracellular free radical damage is an area currently attracting considerable research interest. The compound 2',7'-dichlorofluorescin diacetate (DCFH-DA) is a probe for intracellular peroxide formation commonly used in such studies. During our studies we unexpectedly found that incubation of Trolox, a water soluble vitamin E analog, with DCFH-DA in cell-free...

The DCFH assay is commonly used for measuring free radicals generated by engineered nanomaterials (ENM), a well-established mechanism of ENM toxicity. Concerns exist over susceptibility of the DCFH assay to: assay conditions, adsorption of DCFH onto ENM, fluorescence quenching and light scattering. These effects vary in magnitude depending on ENM physiochemical properties and concentration. A r...

Hydroethidine (HE) and 2',7'-dichlorofluorescin (DCFH) were used for the flow cytometric measurement of reactive oxygen metabolites in leukocytes. Hydroethidine and DCFH were both rapidly oxidized in a cell-free cuvette assay to ethidium bromide (EB) and 2',7'-dichlorofluorescein (DCF) by H2O2 and peroxidase, but not by H2O2 alone, while only HE was oxidized by KO2, a source of O2-. Quiescent l...

PURPOSE To establish a new fluorophotometric method to quantitate oxidative stress in the retina in vivo with a hydrogen peroxide (H2O2)-sensitive fluorescent dye. METHODS For in vitro fluorophotometric study, nonfluorescent 2',7'-dichlorofluorescein (DCFH) was incubated with H2O2 (10 pM to 100 nM), and the production of fluorescent 2',7'-dichlorofluorescein (DCF) was measured with fluorophot...

The ability of murine epidermal cells to produce intracellular hydrogen peroxide was analyzed by flow cytometry and the measurement of 2',7'- dichlorofluorescin (DCFH) oxidation. Epidermal cells isolated from acetone-treated CD-1 mice for 24 h were relatively homogeneous in cell size and density and oxidized low levels of DCFH. However, following 12-O-tetradecanoylphorbol-13-acetate (TPA) treat...

PURPOSE To detect intravitreal peroxides in vivo by a new fluorophotometric method with a hydrogen peroxide (H2O2)-sensitive fluorescent dye. METHODS The authors used a 2',7'-dichlorofluorescin (DCFH) assay to measure oxidative status in the rabbit vitreous. In the presence of H2O2 and lipid hydroperoxides, nonfluorescent DCFH in the vitreous is oxidized to highly fluorescent 2',7'-dichlorofl...

In this study, a new method for detecting creatinine was developed. This novel sensor comprised of two ionic liquids, poly-lactic-co-glycolic acid (PLGA) and 1-butyl-3-methylimidazolium (BMIM) chloride, in the presence of 2',7'-dichlorofluorescein diacetate (DCFH-DA). PLGA and BMIM chloride formed a functional porous polymer structure (FPPS)-like structure. Creatinine within the FPPS rapidly hy...

Understanding the oxidative reactivity of nanoparticles (NPs; <100 nm) could substantially contribute to explaining their toxicity. We attempted to refine the use of 2'7-dichlorodihydrofluorescein (DCFH) to characterize NP generation of reactive oxygen species (ROS). Several fluorescent probes have been applied to testing oxidative reactivity, but despite DCFH being one of the most popular for ...

PURPOSE To develop a highly sensitive method for in vivo quantitation of intravitreal peroxides by vitreous fluorophotometry with 2',7'-dichlorofluorescin (DCFH), a hydrogen peroxide (H2O2)-sensitive fluorescent dye, and to measure peroxides in the vitreous humor after panretinal laser photocoagulation (PRP). METHODS In the presence of H2O2 and lipid hydroperoxides, nonfluorescent DCFH was ox...