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Oroboros |
Description
The Oroboros is the Bioenergetics Platform for high-resolution respirometry (HRR) for mitochondria and cell research, unique for precision OXPHOS analysis.
The Oroboros is available as two models:
Both Oroboros models provide the basis for HRR applications combined with measurement of ROS production, mitochondrial membrane potential, ATP production, Ca2+, pH, and PhotoBiology.
The modular Oroboros concept is supported by DatLab, with high flexibility for add-on Modules. All Oroboros Modules are supported by the Oroboros NextGen-O2k, which is the only model supporting the Redox Modules (Q-Module and NADH-Module), and the NO-Module.
The Oroboros allows bioenergetic analysis at controlled oxygen levels. Small amounts of biological samples can be used for bioenergetic and OXPHOS analysis, ranging from isolated mitochondria, permeabilized tissues and permeabilized cells, to living cells and tissues slices.
The globally tested and trusted high-resolution Oroboros prioritizes both quality and scientific research output in the field of mitochondrial physiology and pathology, extended to PhotoBiology. The Oroboros is a robust sole-source instrument with a long life time and low running costs. Expert service is provided by the Oroboros Open Support.
- » The most common applications of the Oroboros
The Oroboros Bioenergetics Platform - as a driving force in mitochondrial physiology - extends the analytical and diagnostic power of high-resolution respirometry to precision OXPHOS analysis.
The Oroboros is used in basic and applied research to study the respiration of tissues, cells and mitochondria. Fairly simple to highly complex, comprehensive experimental protocols can be applied to assess either basal mitochondrial characteristics or obtain diagnostic insight into respiratory pathways. Topically, studies using the Oroboros range from comparative physiological investigations addressing evolutionary and environmental aspects of mitochondrial metabolism over the energetics related to exercise physiology, nutrition and life style, to aspects of clinical interest such as the impact of metabolic diseases, cancer, or pharmacological interventions.
- The most prominent applications of the Oroboros
- Studies on exercise physiology;nutrition;life style
- » 771 Oroboros Publications
- Diagnosis of acquired and genetic mitochondrial diseases
- Diagnosis of myopathies and neuromuscular pathologies
- Studies on cell function and cell death - apoptosis and necrosis
- Measurement of oxidative stress
- » 586 Oroboros Publications
- Studies on ischemia-reperfusion injury
- » 334 Oroboros Publications
- Studies on aging and senescence
- » 286 Oroboros Publications
- Pharmacological tests
- » 858 Oroboros Publications
- Studies on Diabetes 2
- » 352 Oroboros Publications
- For more applications, browse through the 4814 Oroboros Publications.
Oroboros: development and distribution
- The Oroboros is developed and produced in Austria by Oroboros Instruments in close cooperation with WGT.
- High-resolution respirometry: HRR is the basis for combined measurement of respiration with redox biology (NADH and CoQ), ROS production, mitochondrial membrane potential, ATP production, Ca2+, or pH.
- Titration-Injection microPump TIP2k: The Oroboros 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.
- Oroboros Modules can be combined with the O2k and NextGen-O2k, and can be added any time to extend the Oroboros according to instrument comparability and application-specific objectives: Q (Q-redox state), NAD(P)H autofluorescence (NADH-redox state], PhotoBiology (photosynthesis), FluoRespirometry (ATP, mitochondrial membrane potential, Ca2+), potentiometry (pH, TPP+ or TPMP+ for mitochondrial membrane potential, Ca2+) and amperometry (NO, H2S).
- Auxiliary Oroboros tools provide extended support in the context of high-resolution respirometry: Oxia the O2 regime controller for the Oroboros; Microbalance, Dissection Set.
Bioenergetic profiles, precision OXPHOS analysis, and the Oroboros
- Specific substrate-uncoupler-inhibitor titration (SUIT) protocols are applied in studies of cell respiration or mitochondrial activity. In experiments with living 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 Oroboros MultiSensor modules to simultaneously measure the production of reactive oxygen species or mitochondrial membrane potential, nitric oxide production, acid-base balance or calcium with the Oroboros FluoRespirometer or ion selective electrodes. In the Oroboros, these bioenergetic parameters are always evaluated in combination with the measurement of oxygen concentration and respiration.
- 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 Oroboros 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 Oroboros. 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 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 Oroboros 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 Oroboros in leading O2k-Network Labs are the driving forces for a rapid advancement of mitochondrial physiology. Open access to the Oroboros Manual is an integral part of the concept of Oroboros Instruments on Gentle Science and Scientific Social Responsibility. With sharing of Oroboros Procedures 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.
- Sole source information: pdf
- Frequently Asked Questions
- The Oroboros sets a world-wide standard in high-resolution respirometry - for mitochondrial studies in cell physiology and pathology.
- In contrast to merely offering an oxygen measuring device, the Oroboros is integrated into a systems approach to high-resolution respirometry. This spans from experimental tools which are invaluable in the lab (e.g. Oxia, 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.
What is an O2k and the Oroboros?
- O2k: The Oroboros measures oxygen (dioxygen, O2) as a gas dissolved in aqueous solution.
- With the Oroboros O2k and Oroboros NextGen-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 running.
- An Oxygraph-2k (Oroboros) is used for high-resolution monitoring of respiratory activity of biological or biochemical samples transferred into the experimental chamber. The Oroboros is not used to measure oxygen concentration in a tissue.
- O2k: The O2k and NextGen-O2k has two experimental chambers (k: Kammern, German for chambers). Why 0.5 to 2 mL?
- Power-O2k: Several Oroboros Bioenergetics Platforms are operated simultaneously (P1, P2, ..) to combine high-resolution respirometry with high-output.
- Bioblast links: Oroboros and HRR - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
- The Oroboros Quality Management addresses the acute reproducibility crisis of scientific investigation. In the spirit of Open Science and global networking, we will enable data sharing across projects and institutions in an Open Access database on mitochondrial physiology and pathology, to resolve the inflation crisis and ultimately the value-impact crisis of present academic publication. This supports key developments in mitochondrial medicine. In addition, we expand to algal biotechnology and ecology with the PhotoBiology module of the Oroboros, widening the focus from medicine to environment and climate.
- » Gentle Science recognizes the responsibility of the scientific community - for the quality of life science, the quality of life in science, and its mission.
- » Beyond counting papers – a mission and vision for scientific publication: Bioenerg Commun
- » A vision on preprints
- The Oroboros Quality Management addresses the acute reproducibility crisis of scientific investigation. In the spirit of Open Science and global networking, we will enable data sharing across projects and institutions in an Open Access database on mitochondrial physiology and pathology, to resolve the inflation crisis and ultimately the value-impact crisis of present academic publication. This supports key developments in mitochondrial medicine. In addition, we expand to algal biotechnology and ecology with the PhotoBiology module of the Oroboros, widening the focus from medicine to environment and climate.
Key references - the Oroboros
- Gnaiger E (1983) The twin-flow microrespirometer and simultaneous calorimetry. In: Polarographic Oxygen Sensors. Aquatic and Physiological Applications. Gnaiger E, Forstner H (eds), Springer, Berlin, Heidelberg, New York:134-66.- »Bioblast link«
- Gnaiger E, Steinlechner-Maran R, Méndez G, Eberl T, Margreiter R (1995) Control of mitochondrial and cellular respiration by oxygen. J Bioenerg Biomembr 27:583-96. https://doi.org/10.1007/BF02111656 - »Bioblast link«
- Steinlechner-Maran R, Eberl T, Kunc M, Margreiter R, Gnaiger E (1996) Oxygen dependence of respiration in coupled and uncoupled endothelial cells. Am J Physiol Cell Physiol 271:C2053-61. https://doi.org/10.1152/ajpcell.1996.271.6.C2053- »Bioblast link«
- Gnaiger E, Lassnig B, Kuznetsov AV, Rieger G, Margreiter R (1998) Mitochondrial oxygen affinity, respiratory flux control, and excess capacity of cytochrome c oxidase. https://doi.org/10.1242/jeb.201.8.1129 - »Bioblast link«
- Gnaiger E, Méndez G, Hand SC (2000) High phosphorylation efficiency and depression of uncoupled respiration in mitochondria under hypoxia. Proc Natl Acad Sci U S A 97:11080-5. https://doi.org/10.1073/pnas.97.20.11080 - »Bioblast link«
- Gnaiger E (2001) Bioenergetics at low oxygen: dependence of respiration and phosphorylation on oxygen and adenosine diphosphate supply. Respir Physiol 128:277-97. https://doi.org/10.1016/S0034-5687(01)00307-3 - »Bioblast link«
- Gnaiger E (2003) Oxygen conformance of cellular respiration. A perspective of mitochondrial physiology. Adv Exp Med Biol 543:39-55. https://doi.org/10.1007/978-1-4419-8997-0_4 - »Bioblast link«
- Renner K, Amberger A, Konwalinka G, Gnaiger E (2003) Changes of mitochondrial respiration, mitochondrial content and cell size after induction of apoptosis in leukemia cells. Biochim Biophys Acta 1642:115-23. https://doi.org/10.1016/s0167-4889(03)00105-8 - »Bioblast link«
- Hütter E, Renner K, Pfister G, Stöckl P, Jansen-Dürr P, Gnaiger E (2004) Senescence-associated changes in respiration and oxidative phosphorylation in primary human fibroblasts. Biochem J 380:919-28. https://doi.org/10.1042/BJ20040095 - »Bioblast link«
- Gnaiger E (2008) Polarographic oxygen sensors, the oxygraph and high-resolution respirometry to assess mitochondrial function. In: Mitochondrial dysfunction in drug-induced toxicity (Dykens JA, Will Y, eds) John Wiley & Sons, Inc, Hoboken, NJ:327-52. - »Bioblast link«
- Scandurra FM, Gnaiger E (2010) Cell respiration under hypoxia: facts and artefacts in mitochondrial oxygen kinetics. Adv Exp Med Biol 662:7-25. https://doi.org/10.1007/978-1-4419-1241-1_2 - »Bioblast link«
- Pesta D, Gnaiger E (2012) High-resolution respirometry. OXPHOS protocols for human cells and permeabilized fibers from small biopsies of human muscle. Methods Mol Biol 810:25-58. https://doi.org/10.1007/978-1-61779-382-0_3 - »Bioblast link«
- Harrison DK, Fasching M, Fontana-Ayoub M, Gnaiger E (2015) Cytochrome redox states and respiratory control in mouse and beef heart mitochondria at steady-state levels of hypoxia. J Appl Physiol 119:1210-8. https://doi.org/10.1152/japplphysiol.00146.2015 - »Bioblast link«
- Gnaiger E, Boushel R, Søndergaard H, Munch-Andersen T, Damsgaard R, Hagen C, Díez-Sánchez C, Ara I, Wright-Paradis C, Schrauwen P, Hesselink M, Calbet JAL, Christiansen M, Helge JW, Saltin B (2015) Mitochondrial coupling and capacity of oxidative phosphorylation in skeletal muscle of Inuit and caucasians in the arctic winter. Scand J Med Sci Sports 25 (Suppl 4):126–34. https://doi.org/10.1111/sms.12612 - »Bioblast link«
- Doerrier C, Garcia-Souza LF, Krumschnabel G, Wohlfarter Y, Mészáros AT, Gnaiger E (2018) High-Resolution FluoRespirometry and OXPHOS protocols for human cells, permeabilized fibers from small biopsies of muscle, and isolated mitochondria. Methods Mol Biol 1782:31-70. https://doi.org/10.1007/978-1-4939-7831-1_3 - »Bioblast link«
- Gnaiger E (2020) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 5th ed. Bioenerg Commun 2020.2. https://doi.org/10.26124/bec:2020-0002 - »Bioblast link«
- Komlódi T, Cardoso LHD, Doerrier C, Moore AL, Rich PR, Gnaiger E (2021) Coupling and pathway control of coenzyme Q redox state and respiration in isolated mitochondria. Bioenerg Commun 2021.3. https://doi.org/10.26124/bec:2021-0003 - »Bioblast link«
- Baglivo E, Cardoso LHD, Cecatto C, Gnaiger E (2022) Statistical analysis of instrumental reproducibility as internal quality control in high-resolution respirometry. Bioenerg Commun 2022.8. https://doi.org/10.26124/bec:2022-0008 - »Bioblast link«
- Zdrazilova L, Hansikova H, Gnaiger E (2022) Comparable respiratory activity in attached and suspended human fibroblasts. PLoS ONE 17:e0264496. https://doi.org/10.1371/journal.pone.0264496 - »Bioblast link«
- Baglivo E, Cardoso LHD, Cecatto C, Gnaiger E (2024) Stability of mitochondrial respiration medium used in high-resolution respirometry with living and permeabilized cells. Bioenerg Commun 2024.8. https://doi.org/10.26124/bec.2024-0008 - »Bioblast link«
MitoPedia:
Oroboros hardware,
Oroboros Open Support,
O2k-Respirometry,
O2k-FluoRespirometry
