The oxidation of 2′,7′-dichlorofluorescin to reactive oxygen species: A self-fulfilling prophesy?

Global Profiling of Reactive Oxygen and Nitrogen Species in Biological Systems

“…The fluorescent probes, dichlorodihydrofluorescein (DCFH) and dihydrorhodamine 123 (DHR), have been most widely used to measure intracellular oxidants (H 2 O 2 , ONOO Ϫ ). The pitfalls and artifacts of their use in biological systems have been previously discussed (24,25 (25,26). This is one of the major shortcomings of these probes.…”

Catalase Regulates Cell Growth in HL60 Human Promyelocytic Cells: Evidence for Growth Regulation by H2O2

“…2B), consequently providing a false measurement of intracellular ROS events. Although artifact is minimal because DCFH-DA is substantially more resistant to oxidation than the deacetylated DCFH (38), cells here were irradiated only after they had been fully loaded with DCFH-DA substrate. After 20 min of incubation, over 80% of extracellular DCFH-DA was absorbed by AA8 cells (Fig.…”

The Roles of Mitochondrial Reactive Oxygen Species in Cellular Signaling and Stress Response in Plants

“…Probes such as 3,39-diaminobezidine, nitroblue tetrazolium (NBT), AmplexRed (for isolated mitochondria), 2',7'-dichlorofluorescein derivatives, and hydroethidine have been reported to detect or even quantify H 2 O 2 , superoxide, or mtROS in plant studies (Gleason et al, 2011;Li and Xing, 2011;Martin et al, 2013). However, there are legitimate concerns about their specificity and subcellular accumulation and their interference with endogenous redox biology (Bonini et al, 2006;Zielonka and Kalyanaraman, 2010;Miwa et al, 2016), prompting the search for straightforward and applicable alternatives. Probes that can be specifically placed in mitochondria, either through electrical potential-driven accumulation (e.g.…”