MiPNet06.05 Test Experiments on O2k-Specifications: Difference between revisions
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|title=Gnaiger E ( | |title=[[Image:Logo OROBOROS INSTRUMENTS.jpg|right|60px|link=Oroboros Instruments]]Gnaiger E (2010) Test experiments on specifications of the Oroboros Oxygraph. Mitochondr Physiol Network 6.5. | ||
|info=[[ | |info=[[File:PDF.jpg|100px|link=https://www.bioblast.at/images/6/65/MiPNet06.05_Test_Experiments_on_O2k-Specifications_.pdf|Bioblast pdf]] »[http://www.bioblast.at/index.php/File:MiPNet06.05_O2k-Specifications.pdf Versions] | ||
|authors= | |authors=Oroboros | ||
|year= | |year=2010 | ||
|journal=Mitochondr Physiol Network | |journal=Mitochondr Physiol Network | ||
|abstract=Summary: The | |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 O<sub>2</sub> flux is as low as 0.5 pmol.s<sup>-1</sup>.mL<sup>-1</sup>. Signal noise at zero oxygen concentration is <0.05 μM O<sub>2</sub>. Oxygen backdiffusion at zero oxygen is <3 pmol.s<sup>-1</sup>.mL<sup>-1</sup>, 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. | ||
|mipnetlab=AT Innsbruck | |mipnetlab=AT Innsbruck Oroboros | ||
}} | }} | ||
== 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 |'''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 === | |||
[[File:Flyer backside .png|right|450px]] | |||
:::* 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. | |||
:::::: - PEEK stoppers with Viton O-rings. | |||
:::::: - Electromagnetic stirrers with variable speed (100 to 900 rpm). | |||
:::::: - PVDF-coated stirrer bars (6 mm diameter). | |||
:::::: - Polarographic oxygen sensors sealed with butyl rubber seal tips. | |||
:::* Built-in electronically regulated Peltier thermostat: | |||
:::::: - Temperature range: | |||
::::::{| class="wikitable" | |||
|- | |||
| At room temperature | |||
| 4 °C to 47 °C | |||
|- | |||
| At lower ambient temperature | |||
| 2 °C | |||
|} | |||
:::::: - Temperature stability: | |||
::::::{| class="wikitable" | |||
|- | |||
| Temperature stability | |||
| ±0.002 °C over 90 min. | |||
|} | |||
:::::: - Temperature range: | |||
::::::{| class="wikitable" | |||
|- | |||
| 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: | |||
::::::: Series J and XB | |||
::::::* two additional amperometric signals (O2k-fluorescence; PhotoBiology) | |||
::::::* two additional potentiometric signals (Ion-selective electrodes: for TPP<sup>+</sup>, Ca<sup>2+</sup>, or pH) | |||
::::::: Series XB | |||
::::::* two additional amperometric signals for Q redox state | |||
::::::* two additional amperometric signals (NO) | |||
:::* 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. | |||
:::* internal PC and attached screen | |||
=== Specifications for high-resolution respirometry === | |||
:::* Oxygen signal: | |||
:::::: - Noise at zero O<sub>2</sub>: <±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 O<sub>2</sub>: <±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 O<sub>2</sub> (SD, 100 data points recorded 0.2 s intervals at 37 °C) without smoothing, at 180 µM O<sub>2</sub> (±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). | |||
:::::: - O<sub>2</sub> range of linearity: 0 to 1000 µM. | |||
:::::: - Time constant: <4 s at 37 °C (<3 s typical). | |||
:::* Oxygen flux ''J<sub>O2,V</sub>'' [pmol.s<sup>-1</sup>.mL<sup>-1</sup>]: | |||
:::::: - 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). | |||
:::::: - O<sub>2</sub> range: flux measured up to 500 µM O<sub>2</sub> (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: | |||
:::::: - O<sub>2</sub> backdiffusion [pmol.s<sup>-1</sup>.mL<sup>-1</sup>] at 0 µM: <3 at 20-40 °C (2.5 typical). | |||
:::::: - O<sub>2</sub> backdiffusion [pmol.s<sup>-1</sup>.mL<sup>-1</sup>] at 0 kPa: <3 at 20-40 °C (2.5 typical). | |||
:::::: - O<sub>2</sub> consumption [pmol.s<sup>-1</sup>.mL<sup>-1</sup>] at 200 µM: <4 at 37 °C (3 typical); <3 at 25 °C (2 typical). | |||
:::::: - O<sub>2</sub> consumption [pmol.s<sup>-1</sup>.mL<sup>-1</sup>] at 20 kPa: <4 at 37 °C (3 typical); <3 at 25 °C (2 typical). | |||
{{Labeling | {{Labeling | ||
|area=Instruments;methods | |area=Instruments;methods | ||
|instruments=Oxygraph-2k | |instruments=Oxygraph-2k | ||
}} | }} | ||
[[Category:OroboPedia]] | [[Category:OroboPedia]] |
Latest revision as of 12:49, 31 August 2023
MiPNet06.05 Test Experiments on O2k-Specifications
Gnaiger E (2010) Test experiments on specifications of the Oroboros Oxygraph. Mitochondr Physiol Network 6.5. |
» »Versions
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
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.
- - PEEK stoppers with Viton O-rings.
- - Electromagnetic stirrers with variable speed (100 to 900 rpm).
- - PVDF-coated stirrer bars (6 mm diameter).
- - Polarographic oxygen sensors 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:
- Series J and XB
- two additional amperometric signals (O2k-fluorescence; PhotoBiology)
- two additional potentiometric signals (Ion-selective electrodes: for TPP+, Ca2+, or pH)
- Series XB
- two additional amperometric signals for Q redox state
- two additional amperometric signals (NO)
- - O2k-MultiSensor:
- 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.
- internal PC and attached screen
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).
Labels: MiParea: Instruments;methods
HRR: Oxygraph-2k