Chapter

Oxygen Transport to Tissue XXXIII

Volume 737 of the series Advances in Experimental Medicine and Biology pp 263-268

Date:

Triplet Imaging of Oxygen Consumption During the Contraction of a Single Smooth Muscle Cell (A7r5)

  • Matthias GeissbuehlerAffiliated withLaboratoire d’Optique Biomédicale LOB, École Polytechnique Fédérale de Lausanne (EPFL) Email author 
  • , Thiemo SpielmannAffiliated withLaboratoire d’Optique Biomédicale LOB, École Polytechnique Fédérale de Lausanne (EPFL)Experimental Biomolecular Physics, Department of Applied Physics, Royal Institute of Technology, AlbaNova University Center
  • , Aurélie FormeyAffiliated withLaboratory of Cell Biophysics LCB, École Polytechnique Fédérale de Lausanne (EPFL)
  • , Iwan MärkiAffiliated withLaboratoire d’Optique Biomédicale LOB, École Polytechnique Fédérale de Lausanne (EPFL)
  • , Marcel LeuteneggerAffiliated withLaboratoire d’Optique Biomédicale LOB, École Polytechnique Fédérale de Lausanne (EPFL)Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry
  • , Boris HinzAffiliated withLaboratory of Tissue Repair and Regeneration, CIHR Group in Matrix Dynamics, University of Toronto
  • , Kai JohnssonAffiliated withLaboratory of Protein Engineering LIP1, École Polytechnique Fédérale de Lausanne (EPFL)
  • , Dimitri Van De VilleAffiliated withMedical Image Processing Laboratory MIP, École Polytechnique Fédérale de Lausanne (EPFL)Medical Image Processing Laboratory MIP, University of Geneva
  • , Theo LasserAffiliated withLaboratoire d’Optique Biomédicale LOB, École Polytechnique Fédérale de Lausanne (EPFL)

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Abstract

Triplet imaging is a novel optical technique that allows investigating oxygen metabolism at the single cell and the sub-cellular level. The method combines high temporal and spatial resolutions which are required for the monitoring of fast kinetics of oxygen concentration in living cells. Calibration and validation are demonstrated with a titration experiment using l-ascorbic acid with the enzyme ascorbase oxidase. The method was applied to a biological cell system, employing as reporter a cytosolic fusion protein of β-galactosidase with a SNAP-tag labeled with tetramethylrhodamine. Oxygen consumption in single smooth muscle cells A7r5 during an [Arg8]-vasopressin-induced contraction is measured. The triplet lifetime images over time can be related to an intracellular oxygen consumption corresponding to a mono-exponentially decaying intracellular oxygen concentration. This is in good agreement with previously reported measurements of oxygen consumption in skeletal muscle fibers.