Gnaiger 2008 POS
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Gnaiger E (2008) Polarographic oxygen sensors, the oxygraph and high-resolution respirometry to assess mitochondrial function. In: Mitochondrial Dysfunction in Drug-Induced Toxicity (Dykens JA, Will Y, eds) John Wiley: 327-352. [pdf]
Contents
- Introduction
- Polarographic oxygen sensor and traditional oxygraphy
- High-resolution respirometry: the Oxygraph-2k
- Calibration of polarographic oxygen sensors and oxygen concentration in respiration media at air saturation
- From oxygraph slopes to respiratory flux corrected for background effects
- Phosphorylation control protocol with intact cells
- Titration steps of the PC protocol
- Experimental example for the PC protocol
- Flux control ratios from the PC protocol
- Intact cells, permeabilized cells and tissue, or isolated mitochondria
- Intact cells
- Permeabilized cells and tissue
- Isolated mitochondria
- Titration protocols in permeabilized cells, permeabilized tissue preparations, and isolated mitochondria
- Multisensor applications
Highlights
- Several features distinguish high-resolution respirometry from traditional oxygraphs, combined in the new Oxygraph-2k (O2k, OROBOROS INSTRUMENTS, Innsbruck, Austria; Figure 3). The specifications are unique: the limit of detection of respiratory flux is 1 pmol∙s-1∙cm-3 (0.001 µM∙s-1) and the limit of detection of oxygen concentration extends to 0.005 µM O2. For the non-specialist, the O2k provides robustness and reliability of instrumental performance. With small amounts of sample and correspondingly low respiratory flux per volume, the oxygen capacity of the system is exhausted slowly, allowing sufficient time to evaluate the stability of respiratory activity in each metabolic state and to permit complex titration regimes in intact cells or in permeabilized cells and tissues. To increase throughput in research with cell cultures and in the pharmacological arena, user-friendly features make it possible to apply several instruments in parallel, each O2k with two independent chambers.
