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 & Sons, Inc, Hoboken, NJ:327-52.

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Gnaiger E (2008) John Wiley

Abstract:
Gnaiger 2008 Fig.jpg
Several features distinguish high-resolution respirometry from traditional oxygraphs, combined in the new O2k (O2k, OROBOROS INSTRUMENTS, Innsbruck, Austria). 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.

O2k-Network Lab: AT Innsbruck Gnaiger E, AT Innsbruck OROBOROS


Labels: MiParea: Respiration, Instruments;methods Mammal;model: Mouse Tissue;cell: Blood cells  Preparation: Intact cells, Permeabilized cells, Permeabilized tissue Regulation: Coupling efficiency;uncoupling, Uncoupler Coupling state: LEAK, ROUTINE, ETS Substrate state: ROX HRR: Oxygraph-2k, TIP2k, O2k-Protocol Additional: O2k-Demo, O2k-Core 

Figure legend

Oxygen concentration and oxygen flux in intact cells with coupling control protocol. High-resolution respirometry (OROBOROS O2k with TIP2k) with parental hematopoietic 32D cells at 1.1·106 cells/cm3 suspended in culture medium RPMI at 37 °C. Replicate measurements in the two O2k-chambers (2 cm3). Superimposed plots of oxygen concentration [O2] and volume-specific oxygen flux, JV,O2, calculated as the negative time derivative of oxygen concentration. ROUTINE respiration (R) is followed by inhibition of ATP synthase (manual titration of oligomycin, 2 μg·ml-1) to induce nonphosphorylating LEAK respiration (L). Automatic titration of uncoupler (10 mM FCCP in the TIP2k) in steps of 0.1 μl corresponding to a step increase in the final concentration of 0.5 μM FCCP at intervals of 120 s. Maximum noncoupled flux (capacity of the electron transfer system, [[ETS capacity; state E) is reached at 5.5 μM FCCP. The L/E ratio is 0.10. Respiration is inhibited at higher [FCCP], unrelated to sample dilution (<1%). O2k-experiment 2005-04-09 EF-03, carried out by participants of an O2k-Workshop (Gnaiger 2014 MitoPathways).

» MitoPedia: Respiratory states OXPHOS ROUTINE ETS LEAK - ROX

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