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Talk:Fusion HRR and O2k concept

From Bioblast
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O2k high-resolution respirometry (HRR) is based on the OROBOROS Oxygraph-2k (O2k), which is its modular system, combining chamber design, application of oxygen-tight materials, electrochemical and optical sensors, Peltier-temperature control and software features (DatLab) to obtain the unique sensitive and quantitative resolution of oxygen concentration and oxygen flux, with a closed-chamber or open-chamber mode of operation (TIP2k). Standardized calibration of the polarographic oxygen sensor (static sensor calibration), calibration of the sensor response time (dynamic sensor calibration), and evaluation of instrumental background oxygen flux (systemic flux compensation) provide the experimental basis for high accuracy of quantitative results and quality control in HRR. The most important extension to O2k-MultiSensor systems is the O2k-Fluorometer.

HRR is the result of our long-term expertise in instrumental design, software development, mitochondrial physiology, clinical and pharmacological applications, and world-wide scientific cooperation. The innovative O2k-developments provide a system of highest quality and distinguish HRR as a method and concept.

Today, such high resolution meets rapidly increasing demands, reflected in more than 1,700 O2k-Publications. Applications of the O2k in leading O2k-Network Labs are the driving forces for a rapid advancement of mitochondrial physiology. Open access to the O2k-Manual is an integral part of the concept of OROBOROS INSTRUMENTS on Gentle Science and Scientific Social Responsibility. With sharing of O2k-Protocols and implementation of quality control, the study of mitochondrial and cell respiration emerges as a quantitative science. Thus HRR provides the foundation for comparative mitochondrial physiology, biomedical research, translation into clinical applications, establishing quantitative functional mitochondrial diagnosis.

Gnaiger 1995 J Bioenerg Biomembr, Gnaiger 2008 POS, Gnaiger 2001 Respir Physiol

Dioxygen

An Integrated Systems Approach

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The O2k is the unique state-of-the-art instrument for HRR - where high-resolution counts:

  • with small amounts of cells or tissue
  • at low respiratory activities
  • multiple substrate-uncoupler-inhibitor titration (SUIT protocols) with resolution of changes in oxygen flux over prolonged incubation times
  • flexibility of O2k-MultiSensor applications
  • fast transitions at low oxygen levels
  • stability and quality control

The O2k sets a world-wide standard in HRR - for mitochondrial studies in cell physiology and pathology. Developments progress beyond oxygen.

In contrast to merely offering an oxygen measuring device, the O2k is integrated into a systems approach to HRR. This spans from experimental tools which are invaluable in the lab (e.g. ISS), little aids that are crucial for tissue preparation (e.g. forceps), to information on mitochondrial respiration medium (MiR05 and MiR06), substrate-uncoupler-inhibitor titration protocols, and the unique O2k-Workshops.

What is an Oxygraph?

  • O2k: The O2k measures oxygen (dioxygen, O2) as a gas dissolved in aqueous solution.
  • With the O2k, a graph is displayed not only of oxygen concentration over time, but of respiration (oxygen consumption, oxygen flux) of biological and biochemical samples.

An Oxygraph is not used for measurement of oxygen concentration in a tissue, but for high-resolution analysis of respiratory activity of biological or biochemical samples transferred into the experimental chamber.

The O2k-Core fully supports all O2k-Modules - the Titration-Injection microPump TIP2k and O2k-Multisensor Modules.


MitoPathways
Pesta and Gnaiger 2012

Specific substrate-uncoupler-inhibitor titration (SUIT) protocols are applied in studies of cell respiration or mitochondrial activity. In such experiments, quantitative information is obtained on selected biochemical pathways, particularly of the process of oxidative phosphorylation (OXPHOS) in mitochondria and bacteria. Comprehensive OXPHOS analysis leads us beyond the highly informative process of oxygen consumption, when high-resolution respirometry is combined with O2k-MultiSensor Modules to measure simultaneously the production of reactive oxygen species or mitochondrial membrane potential, nitric oxide production, acid-base balance or calcium with O2k-Fluorometry or ion selective electrodes. In the O2k, these bioenergetic parameters are always evaluated in combination with the measurement of oxygen concentration and respiration.


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  • O2k: The O2k is the 2nd generation of our high-resolution respirometers. The first model was available since 1992. Development of the new O2k started in 2k (2000) and the first O2k was delivered in 2002.


  • O2k: The O2k has two experimental chambers (k: Kammern, German for chambers). Why 2 ml?


  • Power-O2k: Several O2k units are operated simultaneously (P1, P2, ..) to combine high-resolution respirometry with high-ouput.


O2k - modular design

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  • O2k-Core: Substrate-uncoupler-inhibitor titrations (SUIT), OXPHOS flux control (L, P, E), manual titrations, core system for modular add-on extension to use the TIP2k and O2k-MultiSensor Modules.
  • Titration-Injection microPump (TIP2k): The O2k-TIP2k combination allows operation with programmable titration regimes, feedback control, and at steady-states, yielding a new flexibility in experimental design by combining the technical advantages of closed and open systems approaches. Automatic titrations for kinetics; feedback-controlled instrumental background tests; respiration in a closed or open system, in aerobic-anaerobic transitions or at steady-state oxygen concentrations, covering the entire physiological oxygen range and the extremes of hypoxia.
  • O2k-MultiSensor Module: HRR combined with fluorometry (ROS production, ATP production, mitochondrial membrane potential, Ca2+, pH), potentiometry (pH, TPP+ or TPMP+ for mitochondrial membrane potential, Ca2+) and amperometry (NO and oxygen kinetics).
  • Core-Accessories and Auxiliary HRR-Tools are optionally available sets for high-resolution respirometry: Microsyringes, PBI-Shredder, Microbalance, and HRR-Dissection Tools.


Links

Β» MitoPedia: Respiratory states OXPHOS ROUTINE ETS LEAK - ROX