O2 Analysis on a Fluorescence Spectrometer or Plate Reader

  • Alexander V. Zhdanov
  • James Hynes
  • Ruslan I. Dmitriev
  • Dmitri B. Papkovsky
Chapter
First Online:
Part of the SpringerBriefs in Biochemistry and Molecular Biology book series (BRIEFSBIOCHEM)

Abstract

In this chapter, the use of Pt-porphyrin-based extracellular and intracellular O2 sensing probes (ecO2 and icO2) on a TR-F plate reader format is described and critically assessed. The principles underpinning extracellular measurement are outlined and the assessment of biological oxygen consumption in prokaryotic cells, eukaryotic cells and small organisms is outlined along with a description of how such measurements can contribute to the development of a detailed picture of cell metabolism. The use of a mathematical model describing the distribution of local O2 gradients within biological samples is also described. Finally, the principles of icO2 sensing are discussed and some short case studies are provided for demonstrating the utility of icO2 probes for the monitoring of cellular function and metabolic perturbations and how such measurements can be allied to other bioenergetic markers to generate a more complete picture of metabolic status.

Keywords

Oxygen Respiration Cell-based assays Phosphorescent Probes Pt-porphyrins Time-Resolved Fluorescence Oxygen-Sensitive Probes Screening 
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Notes

Acknowledgments

This work was supported by the Science Foundation Ireland, grant 07/IN.1/B1804, and the EU FP7 project Chebana MC-IAPP-2009-230641.

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Copyright information

© The Author(s) 2012

Authors and Affiliations

  • Alexander V. Zhdanov
    • 1
  • James Hynes
    • 2
  • Ruslan I. Dmitriev
    • 1
  • Dmitri B. Papkovsky
    • 1
  1. 1.Biochemistry DepartmentUniversity College CorkCorkIreland
  2. 2.Luxcel Biosciences Ltd. BioInnovation Centre, UCCCorkIreland

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