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Difference between revisions of "MiPNet06.05 Test Experiments on O2k-Specifications"

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{{OROBOROS header page name}}
{{Publication
{{Publication
|title=Gnaiger E (2001) Test experiments on specifications of the OROBOROS Oxygraph. Mitochondr Physiol Network 6.5.
|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=[[Media:MiPNet06.05 O2k-Specifications.pdf|'''Open Access-MiPNet06.05.pdf''']];[http://bioblast.at/index.php?/file:MiPNet06.05_Specifications.pdf];[http://www.oroboros.at/?o2k-specifications www O2k-Specifications]
|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=OROBOROS
|authors=Oroboros
|year=2001
|year=2010
|journal=Mitochondr Physiol Network
|journal=Mitochondr Physiol Network
|abstract=Summary: The OROBOROS Oxygraph 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 pmols-1cm-3. Signal noise at zero oxygen concentration is <0.05 μM O2. Oxygen backdiffusion at zero oxygen is <3 pmols-1cm-3, 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 Oxygraph specifications, which distinguish high-resolution respirometry from any conventional oxygraph system.
|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 OROBOROS
|mipnetlab=AT Innsbruck Oroboros
}}
}}
[[File:Flyer backside .png|right|450px]]
{{Labeling
{{Labeling
|area=Instruments;methods
|area=Instruments;methods
|instruments=Oxygraph-2k
|instruments=Oxygraph-2k
}}
}}
* Version 1: 2001
 
* Version 10: 2010-08-20
== 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 ===
 
:::* 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:
::::::{| 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: 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 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).






[[Category:OroboPedia]]
[[Category:OroboPedia]]

Revision as of 12:46, 24 October 2019

                



MiPNet06.05 Test Experiments on O2k-Specifications

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

» Bioblast pdf »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

Flyer backside .png

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).