 |
Oroboros Procedures |
Oroboros Procedures explain various applications of the Oroboros with Demo experiments.
We aim at a fundamental introduction into high-resolution respirometry and the elements of mitochondrial physiology. This mosaic evolves to combine the elements into a picture of modern mitochondrial respiratory physiology, progressively including user-innovation emerging within the O2k-Network.
| Oroboros Procedures | |
|---|---|
|
» MitoPathways |
 |
» Oroboros Demo experiments |
|
» Oroboros Analysis |
|
» Chemicals and media |
 |
» Mitochondrial preparations |
|
» Mitochondrial and marker-enzymes |
Implement your quality control management with the MitoFit QCS, to diagnose technical problems fast and cost-effectively with instrumental quality control tests.
Oroboros Procedures - with selected chapters of the Oroboros Manual
| Protocol | Reference | Preparation | Tissue and cell | Mammal and model | Last update |
|---|---|---|---|---|---|
| MiPNet22.10 MiR05-Kit |  | 2024-12-12 | |||
| MiPNet21.14 Reference sample HRR |  | Intact cells Permeabilized cells | HEK | Human | 2024-12-06 |
| MiPNet09.12 Manual titrations of SUIT chemicals | | 2024-11-25 | |||
| Donnelly 2024 Redox Biol | Donnelly C, Komlódi T, Cecatto C, Cardoso LHD, Compagnion A-C, Matera A, Tavernari D, Campiche O, Paolicelli RC, Zanou N, Kayser B, Gnaiger E, Place N (2024) Functional hypoxia reduces mitochondrial calcium uptake. Redox Biol 71:103037. https://doi.org/10.1016/j.redox.2024.103037 | Intact organism Isolated mitochondria Intact cells | Heart Skeletal muscle | Human Mouse | 2024 |
| Baglivo 2024 BEC | 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 | Permeabilized cells Intact cells | HEK | Human | 2024 |
| Timon-Gomez 2024 MitoFit | Timón-Gómez A, Doerrier C, Sumbalová Z, Garcia-Souza LF, Baglivo E, Cardoso LHD, Gnaiger E (2024) Analysis of mitochondrial respiratory pathway and coupling control by substrate-uncoupler-inhibitor titration reference protocols. MitoFit Preprints 2024.8. https://doi.org/10.26124/mitofit:2024-0008 | Permeabilized cells Intact cells Homogenate Isolated mitochondria | HEK Heart Nervous system Liver Blood cells | Human Mouse | 2024 |
| Leo 2024 MitoFit | Leo E, Rychtarova L, Garcia-Souza LF, Åsander Frostner E, Elmér E, Gnaiger E (2024) High-resolution respirometry in a small-volume chamber. MitoFit Preprints 2024.4.v2. https://doi.org/10.26124/mitofit:2024-0004.v2 - Accepted on 2024-10-27 as Manuscript JoVE67442R2 for publication in JoVE | Permeabilized cells Homogenate Isolated mitochondria Intact cells | Heart Nervous system Blood cells HUVEC Platelet | Human Mouse | 2024 |
| MiPNet11.05 imt-pti-pce | | Permeabilized cells Permeabilized tissue Isolated mitochondria | 2022-04-10 | ||
| Baglivo 2022 MitoFit-QC | Baglivo E, Cardoso LHD, Cecatto C, Gnaiger E (2022) Statistical analysis of instrumental reproducibility as internal quality control in high-resolution respirometry. https://doi.org/10.26124/mitofit:2022-0018.v2 — 2022-08-04 published in Bioenerg Commun 2022.8. | HEK | 2022 | ||
| Baglivo 2022 BEC | 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 | HEK | 2022 | ||
| MiPNet24.10 H2O2 flux analysis | | 2021-10-22 | |||
| MiPNet26.10 MgG data analysis | | 2021-09-16 | |||
| MiPNet24.09 Data analysis of mt-membrane potential | | 2021-09-16 | |||
| MiPNet20.06 IsolationMouseHeart-mt | | Isolated mitochondria | Heart | Mouse | 2021-08-09 |
| MiPNet17.05 O2k-Fluo LED2-Module | | 2021-06-22 | |||
| Went 2021 MitoFit PB | Went N, Di Marcello M, Gnaiger E (2021) Oxygen dependence of photosynthesis and light-enhanced dark respiration studied by High-Resolution PhotoRespirometry. MitoFit Preprints 2021.05. https://doi.org/10.26124/mitofit:2021-0005 | Intact cells | Algae | 2021 | |
| Cardoso 2021 BEC MgG | Cardoso LHD, Doerrier C, Gnaiger E (2021) Magnesium Green for fluorometric measurement of ATP production does not interfere with mitochondrial respiration. Bioenerg Commun 2021.1. https://doi.org/10.26124/bec:2021-0001 | Isolated mitochondria | Heart | Mouse | 2021 |
| Komlodi 2021 BEC AmR-O2 | Komlódi T, Sobotka O, Gnaiger E (2021) Facts and artefacts on the oxygen dependence of hydrogen peroxide production using Amplex UltraRed. Bioenerg Commun 2021.4. https://doi.org/10.26124/bec:2021-0004 | Intact cells | Other cell lines | Saccharomyces cerevisiae | 2021 |
| Komlodi 2021 BEC Q | 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 | Isolated mitochondria | Heart Nervous system | Mouse | 2021 |
| MiPNet21.17 BloodCellsIsolation | | Intact cells Permeabilized cells | Blood cells Platelet | Human | 2020-10-22 |
| MiPNet24.08 Safranin Analysis Template | | 2020-05-13 | |||
| MiPNet17.04 CitrateSynthase | | Permeabilized cells Permeabilized tissue Homogenate Isolated mitochondria Enzyme | 2020-04-18 | ||
| MiPNet24.06 Oxygen flux analysis - DatLab 7.4 | | 2020-04-16 | |||
| MiPNet25.14 TPP Analysis Template | | 2020-##-## | |||
| BEC 2020.1 doi10.26124bec2020-0001.v1 | Gnaiger E et al ― MitoEAGLE Task Group (2020) Mitochondrial physiology. Bioenerg Commun 2020.1. https://doi.org/10.26124/bec:2020-0001.v1 | Permeabilized cells Permeabilized tissue Homogenate Isolated mitochondria | 2020 | ||
| Gnaiger 2020 BEC MitoPathways | 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 | Permeabilized cells Permeabilized tissue Homogenate Isolated mitochondria Intact cells | Heart Skeletal muscle Fibroblast | Human Mouse | 2020 |
| MiPNet21.16 DatLab 7 Innovations |  | 2019-09-23 | |||
| MiPNet23.12 DL-Protocols help | | 2019-09-13 | |||
| MiPNet22.16 DatLab 7 Innovations: DL-Protocols | | 2019-09-13 | |||
| MiPNet24.11 mtMP calculation | | 2019-08-05 | |||
| MiPNet18.06 Amplex-Yeast | | 2019-06-24 | |||
| MiPNet20.14 AmplexRed H2O2-production | | Isolated mitochondria | Heart | Mouse | 2019-06-24 |
| MiPNet20.13 Safranin mt-membranepotential | | Homogenate | Nervous system | Mouse | 2019-06-24 |
| MiPNet18.05 Amplex-Mouse-heart | | Homogenate | Heart | Mouse | 2019-06-24 |
| Naszai 2019 Sci Rep | Nászai A, Terhes E, Kaszaki J, Boros M, Juhász L (2019) Ca(2+)N it be measured? Detection of extramitochondrial calcium movement with high-resolution FluoRespirometry. Sci Rep 9:19229. | Isolated mitochondria | Liver | Rat | 2019 |
| Long 2019 Methods Mol Biol | Long Q, Huang L, Huang K, Yang Q (2019) Assessing mitochondrial bioenergetics in isolated mitochondria from mouse heart tissues using Oroboros Oxygraph-2k. Methods Mol Biol 1966:237-46. | Isolated mitochondria | Heart | Mouse | 2019 |
| MiPNet12.01 Suppl T-issue | | Permeabilized tissue | Skeletal muscle | Mouse | 2018-11-18 |
| Doerrier 2018 Methods Mol Biol | 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 | Permeabilized cells Permeabilized tissue Homogenate Isolated mitochondria Intact cells | Heart Skeletal muscle Endothelial;epithelial;mesothelial cell Blood cells HEK Platelet | Human Mouse Rat Saccharomyces cerevisiae | 2018 |
| Komlodi 2018 Methods Mol Biol | Komlodi T, Sobotka O, Krumschnabel G, Bezuidenhout N, Hiller E, Doerrier C, Gnaiger E (2018) Comparison of mitochondrial incubation media for measurement of respiration and hydrogen peroxide production. Methods Mol Biol 1782:137-55. https://doi.org/10.1007/978-1-4939-7831-1_8 | Permeabilized cells Permeabilized tissue | Skeletal muscle HEK | Human Mouse | 2018 |
| Lemieux 2017 Sci Rep | Lemieux H, Blier PU, Gnaiger E (2017) Remodeling pathway control of mitochondrial respiratory capacity by temperature in mouse heart: electron flow through the Q-junction in permeabilized fibers. Sci Rep 7:2840. doi:10.1038/s41598-017-02789-8 | Permeabilized tissue | Heart | Mouse | 2017 |
| MiPNet20.08 IsolationRatLiver-mt | | Isolated mitochondria | Liver | Rat | 2016-11-18 |
| MiPNet21.12 Comparison mt respiration media | | Intact cells Permeabilized cells | HEK | Human | 2016-10-18 |
| MiPNet21.13 Comparison mt-preparations | | Permeabilized cells Homogenate Isolated mitochondria | Heart | Mouse | 2016-10-18 |
| MiPNet03.02 Chemicals-Media | Selected media and chemicals for respirometry with mitochondrial preparations. | 2016-08-30 | |||
| MiPNet08.09 CellRespiration | | Intact cells | Blood cells Lymphocyte | Human | 2016-08-11 |
| MiPNet10.04 CellRespiration | | Intact cells | Blood cells | Mouse | 2016-08-08 |
| MiPNet19.18E O2 flux analysis - DatLab 6 | | 2016-08-08 | |||
| MiPNet21.07 MitoFit DatLab PT | | Permeabilized cells | Human | 2016-07-20 | |
| MiPNet20.07 IsolationRatBrain-mt | Laboratory Protocol: isolation of rat brain mitochondria. | Isolated mitochondria | Nervous system | Rat | 2016-06-18 |
| MiPNet14.05 TPP-mtMembranePotential | | 2016-04-01 | |||
| MiPNet20.15 IsolationRatHeart-mt | | Isolated mitochondria | Heart | Rat | 2016-03-25 |
| MiPNet17.15 PBI-Shredder Mouse-heart-brain-liver | | Homogenate | 2015-06-22 | ||
| MiPNet14.14 PermeabilizedFiberPreparation | | Permeabilized tissue | Heart Skeletal muscle | Human | 2015-06-22 |
| MiPNet17.02 PBI-Shredder manual | | Homogenate | 2015-06-19 | ||
| MiPNet17.03 Shredder vs Fibers | | Homogenate | Heart Liver | Fishes | 2015-06-18 |
| MiPNet20.09 IsolationBeefHeart-mt | | Isolated mitochondria | Heart | Bovines | 2015-06-15 |
| Gnaiger 2015 Scand J Med Sci Sports | 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 | Permeabilized tissue | Skeletal muscle | Human | 2015 |
| Makrecka-Kuka 2015 Biomolecules | Makrecka-Kuka M, Krumschnabel G, Gnaiger E (2015) High-resolution respirometry for simultaneous measurement of oxygen and hydrogen peroxide fluxes in permeabilized cells, tissue homogenate and isolated mitochondria. Biomolecules 5:1319-38. https://doi.org/10.3390/biom5031319 | Permeabilized cells Homogenate Isolated mitochondria Intact cells | Heart Nervous system HEK | Human Mouse | 2015 |
| Krumschnabel 2015 Methods Mol Biol | Krumschnabel G, Fontana-Ayoub M, Sumbalova Z, Heidler J, Gauper K, Fasching M, Gnaiger E (2015) Simultaneous high-resolution measurement of mitochondrial respiration and hydrogen peroxide production. Methods Mol Biol 1264:245-61. https://doi.org/10.1007/978-1-4939-2257-4_22 | Isolated mitochondria | Nervous system | Mouse | 2015 |
| MiPNet12.11 MitoRespiration | | Isolated mitochondria | Artemia Crustaceans | 2014-02-20 | |
| Krumschnabel 2014 Methods Enzymol | Krumschnabel G, Eigentler A, Fasching M, Gnaiger E (2014) Use of safranin for the assessment of mitochondrial membrane potential by high-resolution respirometry and fluorometry. Methods Enzymol 542:163-81. https://doi.org/10.1016/B978-0-12-416618-9.00009-1 | Homogenate | Nervous system | Mouse | 2014 |
| Van Bergen 2014 Mitochondrion | Van Bergen NJ, Blake RE, Crowston JG, Trounce IA (2014) Oxidative phosphorylation measurement in cell lines and tissues. Mitochondrion 15:24-33. | Permeabilized cells Isolated mitochondria Intact cells | Nervous system Blood cells | Human Mouse | 2014 |
| Chinopoulos 2014 Methods Enzymol | Chinopoulos C, Kiss G, Kawamata H, Starkov AA (2014) Measurement of ADP-ATP exchange in relation to mitochondrial transmembrane potential and oxygen consumption. Methods Enzymol 542:333-48. https://doi.org/10.1016/B978-0-12-416618-9.00017-0 | Permeabilized cells | HEK | Human | 2014 |
| Elustondo 2014 Biochim Biophys Acta | Elustondo PA, Negoda A, Kane CL, Kane DA, Pavlov EV (2014) Spermine selectively inhibits high-conductance, but not low-conductance calcium-induced permeability transition pore. Biochim Biophys Acta 1847:231–40. | Isolated mitochondria | Liver | Rat | 2014 |
| Pesta 2012 Methods Mol Biol | 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 | Permeabilized cells Permeabilized tissue Intact cells | Skeletal muscle Other cell lines HEK Fibroblast HUVEC | Human | 2012 |
| MiPNet08.12 IOC22 | | Isolated mitochondria | Liver | Rat | 2011-12-11 |
| MiPNet15.08 TPP electrode | | 2011-12-11 | |||
| Lemieux 2011 Int J Biochem Cell Biol | Lemieux H, Semsroth S, Antretter H, Höfer D, Gnaiger E (2011) Mitochondrial respiratory control and early defects of oxidative phosphorylation in the failing human heart. Int J Biochem Cell Biol 43:1729–38. https://doi.org/10.1016/j.biocel.2011.08.008 | Permeabilized tissue | Heart | Human | 2011 |
| MiPNet08.15 Complex-I | | Enzyme | 2010-08-23 | ||
| MiPNet08.18 LactateDehydrogenase | | Enzyme | 2010-08-06 | ||
| MiPNet08.13 mt-Isolation-RLM | | Isolated mitochondria | Liver | Rat | 2010-08-03 |
| Aguirre 2010 Biochim Biophys Acta | Aguirre E, Rodríguez-Juárez F, Bellelli A, Gnaiger E, Cadenas S (2010) Kinetic model of the inhibition of respiration by endogenous nitric oxide in intact cells. Biochim Biophys Acta 1797:557-65. https://doi.org/10.1016/j.bbabio.2010.01.033 | Intact cells Enzyme Oxidase;biochemical oxidation | HEK | 2010 | |
| Gnaiger 2009 Int J Biochem Cell Biol | Gnaiger E (2009) Capacity of oxidative phosphorylation in human skeletal muscle. New perspectives of mitochondrial physiology. Int J Biochem Cell Biol 41:1837-45. https://doi.org/10.1016/j.biocel.2009.03.013 | Permeabilized tissue Isolated mitochondria | Skeletal muscle | Human Mouse | 2009 |
| Gnaiger 2008 POS | 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. | Permeabilized cells Permeabilized tissue Intact cells | Blood cells | Mouse | 2008 |
| Gnaiger 2007 MitoPathways | | 2007 | |||
| Huetter 2004 Biochem J | 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 | Intact cells | Fibroblast | Human | 2004 |
| Gnaiger 2000 Life in the Cold | Gnaiger E, Kuznetsov AV, Schneeberger S, Seiler R, Brandacher G, Steurer W, Margreiter R (2000) Mitochondria in the cold. In: Life in the Cold (Heldmaier G, Klingenspor M, eds) Springer, Berlin, Heidelberg:431-42. https://doi.org/10.1007/978-3-662-04162-8_45 | Permeabilized tissue Isolated mitochondria | Heart | Rat | 2000 |
| MiPNet04.05 Titration-Injection | | Intact cells | Endothelial;epithelial;mesothelial cell HUVEC | Human | 1999-01 |
| Steinlechner-Maran 1996 Am J Physiol Cell Physiol | 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 | Intact cells | Endothelial;epithelial;mesothelial cell HUVEC | Human | 1996 |
