| Analysis of intracellular oxygen and metabolic responses of mammalian cells by time-resolved fluorometry. | |
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MedLine Citation:
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PMID: 18001129 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A simple, minimally invasive methodology for the analysis of intracellular oxygen in populations of live mammalian cells is described. Loading of the cells with the phosphorescent O(2)-sensing probe, MitoXpress, is achieved by passive liposomal transfer or facilitated endocytosis, followed by monitoring in standard microwell plates on a time-resolved fluorescent reader. Phosphorescence lifetime measurements provide accurate, real-time, quantitative assessment of average oxygen levels in resting cells and their alterations in response to stimulation. Analytical performance of the method is examined, optimized, and then demonstrated with different suspension and adherent cell lines including Jurkat, PC12, A549, HeLa, SH-SY5Y, and C2C12, by monitoring responses to mitochondrial uncouplers, inhibitors, plasma membrane depolarization, and Ca(2+) effectors. The assay provides relevant, information-rich data on cellular function and metabolism. It allows monitoring of small, rapid, and transient changes in cell respiration and screening of new chemical entities. |
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Authors:
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Tomas C O'Riordan; Alexander V Zhdanov; Gelii V Ponomarev; Dmitri B Papkovsky |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Validation Studies Date: 2007-11-15 |
Journal Detail:
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Title: Analytical chemistry Volume: 79 ISSN: 0003-2700 ISO Abbreviation: Anal. Chem. Publication Date: 2007 Dec |
Date Detail:
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Created Date: 2007-12-14 Completed Date: 2008-02-21 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0370536 Medline TA: Anal Chem Country: United States |
Other Details:
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Languages: eng Pagination: 9414-9 Citation Subset: IM |
Affiliation:
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Luxcel Biosciences Ltd., BioInnovation Centre, UCC, Cork, Ireland. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Line Cell Respiration Cells / chemistry*, metabolism* Fluorometry / methods* Humans Kinetics Luminescent Measurements Mammals Metabolism Oxygen / analysis* |
| Chemical | |
Reg. No./Substance:
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7782-44-7/Oxygen |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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