| Evaluation of the derivates of phosphorescent Pt-coproporphyrin as intracellular oxygen-sensitive probes. | |
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MedLine Citation:
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PMID: 20063150 Owner: NLM Status: In-Process |
Abstract/OtherAbstract:
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Several new derivatives of the phosphorescent Pt(II)-coproporphyrin (PtCP) were evaluated with respect to the sensing of intracellular oxygen by phosphorescence quenching. Despite the more favorable molecular charge compared to PtCP, self-loading into mammalian cells was rather inefficient for all the dyes, while cell loading by facilitated transport using transfection reagents produced promising results. The PtCP-NH(2) derivative, which gave best loading efficiency and S/N ratio, was investigated in detail including the optimisation of loading conditions, studies of sub-cellular localization, cytotoxicity, oxygen sensitivity and long-term signal stability. Being spectrally similar to the macromolecular MitoXpress™ probe currently used in this application, the PtCP-NH(2) demonstrated higher loading efficiency and phosphorescent signals, suitability for several problematic cell lines and a slightly increased lifetime scale for the physiological range (0-200 μM O(2)). In physiological experiments with different cell types, mitochondrial uncouplers and inhibitors performed on a time-resolved fluorescence plate reader, this probe produced the anticipated profiles of intracellular oxygen concentration and responses to cell stimulation. Therefore, PtCP-NH(2) represents a convenient probe for the experiments and applications in which monitoring of cellular oxygen levels is required. |
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Authors:
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Andreas Fercher; Gelii V Ponomarev; Dmitri Yashunski; Dmitri Papkovsky |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-01-10 |
Journal Detail:
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Title: Analytical and bioanalytical chemistry Volume: 396 ISSN: 1618-2650 ISO Abbreviation: Anal Bioanal Chem Publication Date: 2010 Mar |
Date Detail:
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Created Date: 2010-09-30 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101134327 Medline TA: Anal Bioanal Chem Country: Germany |
Other Details:
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Languages: eng Pagination: 1793-803 Citation Subset: IM |
Affiliation:
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Biochemistry Department, Cavanagh Building, University College Cork, Cork, Ireland. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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