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Oroboros O2k - the modular system for high-resolution respirometry (HRR) for mitochondria and cell research. The O2k-Core (O2k-Series D - G with the O2k-Fluo LED2-Module) and the O2k-FluoRespirometer (O2k-Series H) are the basis for combined measurement of respiration and fluorometric signals.
Small amounts of biological samples can be used for bioenergetic and OXPHOS analysis, ranging from isolated mitochondria, permeabilized tissues and permeabilized cells to intact cells and tissues slices.
The modular O2k-concept is supported by DatLab 7 (new), with high flexibility for extension by add-on O2k-Modules or economical first steps with the Startup O2k-Respirometer. The high-resolution O2k technology is world-wide tested and trusted with emphasis on quality and scientific output of research in mitochondrial physiology and pathology.
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Oroboros O2k: development and distribution
- High-resolution respirometry: HRR is the basis for combined measurement of respiration and ROS production, mitochondrial membrane potential, ATP production, Ca2+, or pH.
- 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.
What is an Oxygraph and the O2k?
- O2k: The O2k measures oxygen (dioxygen, O2) as a gas dissolved in aqueous solution.
- With the O2k, experimental data are displayed not only of oxygen concentration over time; but real-time transformation of these data allows the experimenter to follow respiration (oxygen consumption, oxygen flux) of biological and biochemical samples while the experiment is still running.
- An Oxygraph is not used to measure oxygen concentration in a tissue, but for high-resolution analysis of respiratory activity of biological or biochemical samples transferred into the experimental chamber.
- Specific substrate-uncoupler-inhibitor titration (SUIT) protocols are applied in studies of cell respiration or mitochondrial activity. In experiments with intact cells, different coupling control states can be studied to evaluate ROUTINE respiratory activity, respiration coupled to ATP production, respiratory excess capacity, residual oxygen consumption, and, by extension of conventional protocols, intactness of cell membranes. In mitochondrial preparations such as isolated mitochondria, permeabilized cells and tissue samples, or homogenized tissue, quantitative information is obtained on selected mitochondrial 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 simultaneously measure the production of reactive oxygen species or mitochondrial membrane potential, nitric oxide production, acid-base balance or calcium with O2k-FluoRespirometry or ion selective electrodes. In the O2k, these bioenergetic parameters are always evaluated in combination with the measurement of oxygen concentration and respiration.
Oroboros O2k: O2k and high-resolution respirometry
- 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-output.
- High-resolution respirometry (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.
- HRR is defined on the basis of instrumental features of the O2k. The limit of detection of oxygen flux is as low as 1 pmol O2.s-1.mL-1. Oxygen backdiffusion at zero oxygen is <4 pmol.s-1.mL-1. Oxygen consumption by the polarographic oxygen sensor at air saturation and standard barometric pressure (100 kPa) is 2.7 pmol.s-1.mL-1 (± 0.9 SD; 114 test runs at 37 °C). These highly standardized instrumental background fluxes are a linear function of oxygen concentration, which is used for automatic background correction of oxygen flux (DatLab). Typical exponential time constants of the oxygen sensors are <4 s, used for dynamic corrections in kinetic studies. Signal noise at zero oxygen concentration is <0.05 µM O2.
- The Oroboros 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 titrations (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.
- Today, such high resolution meets rapidly increasing demands, reflected by the increasing number and high level of 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 high-resolution respirometry provides the foundation for comparative mitochondrial physiology, biomedical research, translation into clinical applications, establishing quantitative functional mitochondrial diagnosis.
- The Oroboros O2k sets a world-wide standard in high-resolution respirometry - for mitochondrial studies in cell physiology and pathology. Developments progress beyond oxygen.
- In contrast to merely offering an oxygen measuring device, the Oroboros O2k is integrated into a systems approach to high-resolution respirometry. This spans from experimental tools which are invaluable in the lab (e.g. ISS), or little aids that are crucial for tissue preparation (forceps), mitochondrial respiration medium (MiR05-Kit), substrate-uncoupler-inhibitor titration protocols, and the unique O2k-Workshops.
O2k-FluoRespirometer: an integrated systems approach
- The O2k-FluoRespirometer is the basis for combined measurement of respiration and fluorometric signals, supported by the Titration-Injection microPump and DatLab 7 (new). The modular O2k-concept yields a high flexibility of FluoRespirometry with add-on O2k-Modules for Mitochondria and Cell Research.