| Triplet imaging of oxygen consumption during the contraction of a single smooth muscle cell (A7r5). | |
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MedLine Citation:
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PMID: 20338856 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The measurement of tissue and cell oxygenation is important for understanding cell metabolism. We have addressed this problem with a novel optical technique, called triplet imaging, that exploits oxygen-induced triplet lifetime changes and is compatible with a variety of fluorophores. A modulated excitation of varying pulse widths allows the extraction of the lifetime of the essentially dark triplet state using a high-fluorescence signal intensity. This enables the monitoring of fast kinetics of oxygen concentration in living cells combined with high temporal and spatial resolution. First, the oxygen-dependent triplet-state quenching of tetramethylrhodamine is validated and then calibrated in an L-ascorbic acid titration experiment demonstrating the linear relation between triplet lifetime and oxygen concentration according to the Stern-Volmer equation. Second, the method is applied to a biological cell system, employing as reporter a cytosolic fusion protein of beta-galactosidase with SNAP-tag labeled with tetramethylrhodamine. Oxygen consumption in single smooth muscle cells A7r5 during an [Arg(8)]-vasopressin-induced contraction is measured. The results indicate a consumption leading to an intracellular oxygen concentration that decays monoexponentially with time. The proposed method has the potential to become a new tool for investigating oxygen metabolism at the single cell and the subcellular level. |
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Authors:
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Matthias Geissbuehler; Thiemo Spielmann; Aurélie Formey; Iwan Märki; Marcel Leutenegger; Boris Hinz; Kai Johnsson; Dimitri Van De Ville; Theo Lasser |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Validation Studies |
Journal Detail:
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Title: Biophysical journal Volume: 98 ISSN: 1542-0086 ISO Abbreviation: Biophys. J. Publication Date: 2010 Jan |
Date Detail:
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Created Date: 2010-03-26 Completed Date: 2010-06-17 Revised Date: 2011-07-20 |
Medline Journal Info:
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Nlm Unique ID: 0370626 Medline TA: Biophys J Country: United States |
Other Details:
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Languages: eng Pagination: 339-49 Citation Subset: IM |
Copyright Information:
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Copyright 2010 Biophysical Society. Published by Elsevier Inc. All rights reserved. |
Affiliation:
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Laboratoire d'Optique Biomédicale, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland. matthias.geissbuehler@epfl.ch |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Algorithms Animals Arginine Vasopressin / pharmacology Ascorbic Acid / chemistry Calibration Cell Line Fluorescence* Image Processing, Computer-Assisted Intracellular Space / metabolism Kinetics Linear Models Models, Chemical Muscle Contraction / drug effects, physiology* Myocytes, Smooth Muscle / drug effects, physiology* Optics and Photonics / methods* Oxygen / chemistry, metabolism Oxygen Consumption* Rats Rhodamines / chemistry Vasoconstrictor Agents / pharmacology Video Recording beta-Galactosidase / chemistry, metabolism |
| Chemical | |
Reg. No./Substance:
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0/Rhodamines; 0/Vasoconstrictor Agents; 113-79-1/Arginine Vasopressin; 50-81-7/Ascorbic Acid; 62669-72-1/tetramethylrhodamine; 7782-44-7/Oxygen; EC 3.2.1.23/beta-Galactosidase |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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