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Triplet Imaging of ...
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Geissbuehler, Matthias
(författare)
Triplet Imaging of Oxygen Consumption during the Contraction of a Single Smooth Muscle Cell (A7r5)
- Artikel/kapitelEngelska2010
Förlag, utgivningsår, omfång ...
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Elsevier BV,2010
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Nummerbeteckningar
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LIBRIS-ID:oai:DiVA.org:kth-28879
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https://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-28879URI
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https://doi.org/10.1016/j.bpj.2009.10.006DOI
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Språk:engelska
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Sammanfattning på:engelska
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Ämneskategori:ref swepub-contenttype
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Ämneskategori:art swepub-publicationtype
Anmärkningar
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QC 20110207
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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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Biuppslag (personer, institutioner, konferenser, titlar ...)
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Spielmann, ThiemoKTH,Experimentell biomolekylär fysik(Swepub:kth)u176i2oc
(författare)
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Formey, Aurlie
(författare)
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Maerki, Iwan
(författare)
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Leutenegger, Marcel
(författare)
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Hinz, Boris
(författare)
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Johnsson, Kai
(författare)
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Van De Ville, Dimitri
(författare)
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Lasser, Theo
(författare)
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KTHExperimentell biomolekylär fysik
(creator_code:org_t)
Sammanhörande titlar
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Ingår i:Biophysical Journal: Elsevier BV98:2, s. 339-3490006-34951542-0086
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