Evaluation of different methods detecting intracellular generation of free radicals
Language English Country Netherlands Media print-electronic
Document type Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Epithelial Cells metabolism MeSH
- Ethidium analogs & derivatives MeSH
- Fluoresceins MeSH
- Rabbits MeSH
- Rats MeSH
- Methods MeSH
- Molecular Probe Techniques * MeSH
- Myocardium metabolism MeSH
- Lens, Crystalline cytology MeSH
- Reactive Oxygen Species analysis MeSH
- Rhodamines MeSH
- Free Radicals analysis MeSH
- Animals MeSH
- Check Tag
- Rabbits MeSH
- Rats MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 2',7'-dichlorofluorescein MeSH Browser
- dihydroethidium MeSH Browser
- dihydrorhodamine 123 MeSH Browser
- Ethidium MeSH
- Fluoresceins MeSH
- Reactive Oxygen Species MeSH
- Rhodamines MeSH
- Free Radicals MeSH
Reactive oxygen species (ROS) play several biological roles. We investigated the applicability of fluorescent probes for their detection (i) in rabbit lens epithelial cells during ageing in culture, and (ii) in thin sections of rat heart. We used dihydroethidium (DHE), dichlorofluorescin (DCFH), and dihydrorhodamine 123 (DHR) together with detection of autofluorescence both in cells and in chloroform extracts. Superoxide production was confirmed by a specific histochemical method using Mn(2+). All methods demonstrated higher production of ROS in older cells. All probes revealed different sites of ROS production in young and old cells and could be used for investigation of ROS generation during cell ageing. In the thin sections of rat heart DCFH was not suitable for intracellular ROS detection. The results indicate that the potential of fluorescent dyes in ROS detection is not usually fully exploited, and that blue autofluorescence is associated with oxidative damage.
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