Fluorescence Imaging and Flow Cytometric Analysis of Hydrogen Peroxide in Living Cells Using a Novel Fluorescent Probe

As a type of reactive oxygen species (ROS), hydrogen peroxide (H2O2) is a reactive oxygen metabolic by-product that serves as a key regulator for a number of oxidative stress-related states. It is involved in many biological events that are linked to asthma, atherosclerosis, diabetic vasculopathy, osteoporosis, a number of neurodegenerative diseases and Down’s syndrome. The measurement of this reactive species is helpful for determining how oxidative stress modulates various intracellular pathways. However, fluorescent probes which can detect H2O2 produced in mitochondrial remain rare. OxiVision Blue was developed to detect mitochondrial H2O2 in living cells. Cervial cancer HeLa cells stained with the novel fluorescent probe show negligible fluorescence in absence of peroxide stimulants. In contrast, the treatment of OxiVision Blue -loaded cells with H2O2 displayed strong blue fluorescence in mitochondria. The probe has also been successfully applied in flow cytometer for the quantitative detection of H2O2 in individual cells. Taken together, these results demonstrate that OxiVision Blue is mitochondria-targeted H2O2 probe, and the exogenous and endogenous changes of H2O2 levels in living cells can be monitored in real time and quantitatively measured by fluorescence imaging and flow cytometer.