Document Detail

Evaluation of different methods detecting intracellular generation of free radicals.
MedLine Citation:
PMID:  19301099     Owner:  NLM     Status:  MEDLINE    
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.
Jirí Wilhelm; Richard Vytásek; Ivana Ostádalová; Ludĕk Vajner
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-03-20
Journal Detail:
Title:  Molecular and cellular biochemistry     Volume:  328     ISSN:  1573-4919     ISO Abbreviation:  Mol. Cell. Biochem.     Publication Date:  2009 Aug 
Date Detail:
Created Date:  2009-07-01     Completed Date:  2009-11-03     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0364456     Medline TA:  Mol Cell Biochem     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  167-76     Citation Subset:  IM    
Department of Medical Chemistry and Biochemistry, Centre of Cardiovascular Research, Charles University, Prague, Czech Republic.
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MeSH Terms
Epithelial Cells / metabolism
Ethidium / analogs & derivatives
Free Radicals / analysis*
Lens, Crystalline / cytology
Molecular Probe Techniques*
Myocardium / metabolism
Reactive Oxygen Species / analysis*
Reg. No./Substance:
0/Fluoresceins; 0/Free Radicals; 0/Reactive Oxygen Species; 0/Rhodamines; 104821-25-2/dihydroethidium; 109244-58-8/dihydrorhodamine 123; 3546-21-2/Ethidium; 76-54-0/2',7'-dichlorofluorescein

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