| Simultaneous analysis of reactive oxygen species and reduced glutathione content in living cells by polychromatic flow cytometry. | |
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MedLine Citation:
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PMID: 20010930 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Reactive oxygen species (ROS) are continuously produced in the cell as a consequence of aerobic metabolism, and are controlled by several antioxidant mechanisms. An accurate measurement of ROS is essential to evaluate the redox status of the cell, or the effects of molecules with the pro-oxidant or antioxidant activity. Here we report a cytofluorimetric technique for measuring simultaneously, at the single-cell level, hydrogen peroxide and superoxide anion, reduced glutathione (a main intracellular antioxidant) and cell viability. The staining is performed with the fluorescent dyes 2',7'-dichlorodihydrofluorescein diacetate (H2DCFH-DA), hydroethidine (HE), monobromobimane (MBB) and TO-PRO-3. This analysis is possible with new-generation flow cytometers equipped with several light sources (in our case, four lasers and an UV lamp), which excite different fluorochromes. This approach is extremely useful to study the balance between ROS content and antioxidants in cells receiving different stimuli, and to analyze the relationship between oxidative stress and cell death. |
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Authors:
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Andrea Cossarizza; Roberta Ferraresi; Leonarda Troiano; Erika Roat; Lara Gibellini; Linda Bertoncelli; Milena Nasi; Marcello Pinti |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Nature protocols Volume: 4 ISSN: 1750-2799 ISO Abbreviation: Nat Protoc Publication Date: 2009 |
Date Detail:
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Created Date: 2009-12-16 Completed Date: 2010-03-05 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101284307 Medline TA: Nat Protoc Country: England |
Other Details:
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Languages: eng Pagination: 1790-7 Citation Subset: IM |
Affiliation:
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Department of Biomedical Sciences, University of Modena and Reggio Emilia, Modena, Italy. andrea.cossarizza@unimore.it |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Cell Line Flow Cytometry / instrumentation, methods* Fluorescent Dyes / analysis Glutathione / metabolism* Humans Hydrogen Peroxide / metabolism Oxidation-Reduction Reactive Oxygen Species / metabolism* Superoxides / metabolism |
| Chemical | |
Reg. No./Substance:
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0/Fluorescent Dyes; 0/Reactive Oxygen Species; 11062-77-4/Superoxides; 70-18-8/Glutathione; 7722-84-1/Hydrogen Peroxide |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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