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

Abstract

As a type of reactive oxygen species (ROS), hydrogen peroxide (H 2 O 2 ) 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 H 2 O 2 produced in mitochondrial remain rare. OxiVision Blue was developed to detect mitochondrial H 2 O 2 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 H 2 O 2 displayed strong blue fluorescence in mitochondria. The probe has also been successfully applied in flow cytometer for the quantitative detection of H 2 O 2 in individual cells. Taken together, these results demonstrate that OxiVision Blue is mitochondria-targeted H 2 O 2 probe, and the exogenous and endogenous changes of H 2 O 2 levels in living cells can be monitored in real time and quantitatively measured by fluorescence imaging and flow cytometer.