| Imaging of cellular oxygen and analysis of metabolic responses of mammalian cells. | |
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MedLine Citation:
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PMID: 19957136 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Many parameters reflecting mitochondrial function and metabolic status of the cell, including the mitochondrial membrane potential, reactive oxygen species, ATP, NADH, ion gradients, and ion fluxes (Ca(2+), H(+)), are amenable for analysis by live cell imaging and are widely used in many labs. However, one key metabolite - cellular oxygen - is currently not analyzed routinely. Here we present several imaging techniques that use the phosphorescent oxygen-sensitive probes loaded intracellularly and which allow real-time monitoring of O(2) in live respiring cells and metabolic responses to cell stimulation. The techniques include conventional wide-field fluorescence microscopy to monitor relative changes in cell respiration, microsecond FLIM format which provides quantitative readout of O(2) concentration within/near the cells, and live cell array devices for the monitoring of metabolic responses of individual suspension cells. Step by step procedures of typical experiments for each of these applications and troubleshooting guide are given. |
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Authors:
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Andreas Fercher; Tomas C O'Riordan; Alexander V Zhdanov; Ruslan I Dmitriev; Dmitri B Papkovsky |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Methods in molecular biology (Clifton, N.J.) Volume: 591 ISSN: 1940-6029 ISO Abbreviation: Methods Mol. Biol. Publication Date: 2010 |
Date Detail:
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Created Date: 2009-12-03 Completed Date: 2010-02-18 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9214969 Medline TA: Methods Mol Biol Country: United States |
Other Details:
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Languages: eng Pagination: 257-73 Citation Subset: IM |
Affiliation:
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Biochemistry Department, University College Cork, Cork, Ireland. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Culture Techniques Cell Differentiation Cell Line Cell Survival Fluorescent Dyes / metabolism Humans Intracellular Space / metabolism Mitochondria / metabolism Molecular Imaging / methods* Organoplatinum Compounds / metabolism Oxygen / metabolism* Spectrometry, Fluorescence |
| Chemical | |
Reg. No./Substance:
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0/Fluorescent Dyes; 0/Organoplatinum Compounds; 7782-44-7/Oxygen |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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