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MiPNet06.05 Test Experiments on O2k-Specifications

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MiPNet06.05 Test Experiments on O2k-Specifications

Publications in the MiPMap
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Gnaiger E (2010) Test experiments on specifications of the Oroboros Oxygraph. Mitochondr Physiol Network 6.5.

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Oroboros (2010) Mitochondr Physiol Network

Abstract: Summary: The Oroboros O2k with DatLab software is world-wide the only instrument on the market, which allows routine measurements to be made with specifications summarized under the term “high-resolution respirometry”: The limit of detection of O2 flux is as low as 0.5 pmol.s-1.ml-1. Signal noise at zero oxygen concentration is <0.05 μM O2. Oxygen backdiffusion at zero oxygen is <3 pmol.s-1.ml-1, and oxygen consumption at air saturation and standard barometric pressure (100 kPa) was 2.7 +/- 0.9 SD in 114 test runs at 37 °C. These highly standardized instrumental background fluxes are a linear function of oxygen concentration, which is used for routine background correction of oxygen flux. Typical exponential time constants of the oxygen sensors are <4 s, used for dynamic corrections in kinetic studies. Results of an extensive experimental test program are presented as the basis of Oroboros O2k-Specifications, which distinguish high-resolution respirometry from any conventional oxygraph system.


O2k-Network Lab: AT Innsbruck Oroboros

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Labels: MiParea: Instruments;methods 





HRR: Oxygraph-2k 



O2k High-resolution respirometry

The Oroboros O2k with DatLab software is world-wide the only instrument (sole source) which allows routine measurements to be made with specifications summarized under the term high-resolution respirometry (HRR).
Continuous control of the Oroboros O2k-Specifications are a fundamental component of Quality Assurance and distinguish high-resolution respirometry from any conventional oxygraph system and from less quantitative approaches.
» O2k-Specifications


Oroboros O2k: technical specifications

General specifications

  • Dimensions - O2k-Main Unit: L 45 cm, W 31 cm, H 25 cm; 13.45 kg without packing box or Peli Case.
  • Chambers: chemically inert and minimum oxygen diffusion:
- Two Duran glass chambers with 2 ml (1.5 to 3.5 ml) volume; 16 mm inner diameter.
- PVDF or PEEK stoppers with Viton O-rings.
- Electromagnetic stirrers with variable speed (100 to 900 rpm).
- PVDF- or PEEK-coated stirrer bars (6 mm diameter).
- Polarographic oxygen sensors (OroboPOS) sealed with butyl rubber seal tips.
  • Built-in electronically regulated Peltier thermostat:
- Temperature range:
At room temperature 4 °C to 47 °C
At lower ambient temperature 2 °C
- Temperature stability:
Temperature stability ±0.002 °C over 90 min.
- Temperature range:
in 15 min 20 °C to 30 °C
in 20 min 30 °C to 20 °C
  • Data output through USB:
- Two oxygen signals.
- Pressure transducer, for absolute barometric pressure signal, resolution 0.1 kPa.
- Thermostat temperature signal, resolution 0.001 °C.
- O2k-MultiSensor: two additional amperometric signals (O2k-fluorescence; NO sensors), and two additional potentiometric signals pX: for TPP or pH).
  • DatLab software:
- Simultaneous display of O2 concentration and O2 flux (negative slope or time derivative), together with any selected O2k-MultiSensor channels (raw or calibrated signals and corresponding time derivatives).
- Barometric pressure for O2 calibration; temperature stability control.
- Automatic calibrations: O2 and O2k-MultiSensor channels.
- DL-Protocols: real-time guide through instrumental quality control tests and advanced diagnostic substrate-uncoupler-inhibitor titration (SUIT) protocols within a single assay for exploring mitochondrial function.



Specifications for high-resolution respirometry

  • Oxygen signal:
- Noise at zero O2: <±0.02 µM (SD, 100 data points recorded at 0.2 s intervals at 37 °C) without smoothing (±0.005 µM typical)
- Noise at zero O2: <±0.002 kPa (SD, 100 data points recorded at 0.2 s intervals) without smoothing (±0.0005 kPa typical).
- Noise at air saturation: <±0.1 µM O2 (SD, 100 data points recorded 0.2 s intervals at 37 °C) without smoothing, at 180 µM O2 (±0.05 µM typical).
- Noise at air saturation: <±0.01 kPa (SD, 100 data points recorded 0.2 s intervals) without smoothing, at partial oxygen pressure of 20 kPa (±0.005 kPa typical).
- O2 range of linearity: 0 to 1000 µM.
- Time constant: <4 s at 37 °C (<3 s typical).


  • Oxygen flux JO2,V [pmol.s-1.ml-1]:
- Limit of detection: 0.5 at steady-state over 5 min.
- Sensitivity (normoxic): <1 at steady-state over 5 min at 20-40 °C.
- Sensitivity (hyperoxic): <3 at steady-state over 5 min at 20-40 °C.
- Noise: <0.2 after standard smoothing (120 s).
- O2 range: flux measured up to 500 µM O2 (permeabilized fibres), and <0.1 µM based on DatLab analysis of oxygen kinetics (mitochondria and cells).


  • Instrumental background for linear correction over the entire oxygen range:
- O2 backdiffusion [pmol.s-1.ml-1] at 0 µM: <3 at 20-40 °C (2.5 typical).
- O2 backdiffusion [pmol.s-1.ml-1] at 0 kPa: <3 at 20-40 °C (2.5 typical).
- O2 consumption [pmol.s-1.ml-1] at 200 µM: <4 at 37 °C (3 typical); <3 at 25 °C (2 typical).
- O2 consumption [pmol.s-1.ml-1] at 20 kPa: <4 at 37 °C (3 typical); <3 at 25 °C (2 typical).