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Title

Triplet imaging of oxygen consumption during the contraction of a single smooth muscle cell (A7r5)

Authors
Geissbuehler, Matthias
Spielmann, Thiemo
Formey, Aurelie
Marki, Iwan
Leutenegger, Marcel
Johnsson, Kai
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Published in Biophysical Journal. 2010, vol. 98, no. 2, p. 339-349
Abstract 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.
Keywords AlgorithmsAnimalsArginine Vasopressin/pharmacologyAscorbic Acid/chemistryCalibrationCell Line*FluorescenceImage Processing, Computer-AssistedIntracellular Space/metabolismKineticsLinear ModelsModels, ChemicalMuscle Contraction/drug effects/*physiologyMyocytes, Smooth Muscle/drug effects/*physiologyOptics and Photonics/*methodsOxygen/chemistry/metabolism*Oxygen ConsumptionRatsRhodamines/chemistryVasoconstrictor Agents/pharmacologyVideo RecordingBeta-Galactosidase/chemistry/metabolism
Identifiers
PMID: 20338856
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Research group Traitement d'images médicales (893)
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GEISSBUEHLER, Matthias et al. Triplet imaging of oxygen consumption during the contraction of a single smooth muscle cell (A7r5). In: Biophysical Journal, 2010, vol. 98, n° 2, p. 339-349. doi: 10.1016/j.bpj.2009.10.006 https://archive-ouverte.unige.ch/unige:20957

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Deposited on : 2012-05-23

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