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Real-time measurement of intracellular reactive oxygen species using Mito tracker orange (CMH2TMRos)
- Source :
- Bioscience reports. 21(3)
- Publication Year :
- 2002
-
Abstract
- We have investigated a novel method to monitor real changes of intracellular ROS by the use of CMH2TMRos (a reduced form of MitoTracker orange) in Swiss 3T3 fibroblasts. Arachidonic acid induced a rapid increase of CMTMRos fluorescence with a maximal elevation at 120–150 sec, which was determined by scanning every 10 sec with a confocal microscope. The fluorescence increase by arachidonic acid was completely inhibited by 2-MPG but not by catalase, indicating a major contribution of superoxide to the oxidation of CMH2TMRos. Incubation with glucose oxidase, exogenous H2O2, KO2 and lysophosphatidic acid also increased the CMTMRos fluorescence, which was blocked by 2-MPG. These results suggested that CMH2TMRos is a useful fluorophore for real-time monitoring of intracellular ROS and also indicated that CMH2TMRos detects primarily superoxide in cells even though the fluorophore can be oxidized by both superoxide and H2O2.
- Subjects :
- Intracellular Fluid
Fluorophore
Time Factors
Biophysics
Biochemistry
chemistry.chemical_compound
Mice
Lysophosphatidic acid
Animals
Glucose oxidase
Molecular Biology
Fluorescent Dyes
chemistry.chemical_classification
Reactive oxygen species
Arachidonic Acid
Microscopy, Confocal
biology
Superoxide
Histocytochemistry
Tiopronin
Cell Biology
3T3 Cells
Catalase
Mitochondria
Eukaryotic Cells
chemistry
Xanthenes
biology.protein
Arachidonic acid
Lysophospholipids
Reactive Oxygen Species
Oxidation-Reduction
Intracellular
Subjects
Details
- ISSN :
- 01448463
- Volume :
- 21
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- Bioscience reports
- Accession number :
- edsair.doi.dedup.....ebdf3adb14a2386f53a61445f04405e9