NextGen-O2k

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NextGen-O2k


NextGen-O2k         K-Regio MitoFit         O2k-Innovation         Houska-Award 2012         K-Regio MitoCom         Bioblast 2012


NextGen-O2k

The revolutionary all-in-one instrument to conquer mitochondrial disease

NextGen-O2k


NextGen-O2k: Horizon 2020 Framework Programme

  • EIC-SMEInst-2018-2020 — SME instrument phase 2, Grant Agreement No. 859770, Oroboros Instruments
  • Budget: € 2.35 Million
  • Duration: 24 months, extended additional 5 months
  • Application: 2019-01-09; Pitch in Brussels: 2019-02-15; Granted: 2019-03-05; Start: 2019-06-01


NextGen-O2k publications, preprints, and abstracts

LinkReferenceViewYear
O2k-Protocols
NextGen-O2k: Q-Module manual
PDF.jpg » Versions2021-01-27
Went N, Di Marcello M, Gnaiger E (2021) Oxygen dependence of photosynthesis and light-enhanced dark respiration studied by High-Resolution PhotoRespirometry. MitoFit Prep 2021.5. doi:10.26124/mitofit:2021-0005
MitoFit pdf
Oxygen dependence of photosynthesis and light-enhanced dark respiration studied by High-Resolution PhotoRespirometry
2021-04-30
Ravasz D, Bui D, Kitayev A, Greenwood B, Hill C, Komlodi T, Doerrier C, Ozohanics O, Moore AL, Gnaiger E, Kiebish M, Kolev K, Seyfried TN, Willis WT, Narain N, Adam-Vizi V, Chinopoulos C (2019) Endogenous quinones sustain a moderate NADH oxidation by Complex I during anoxia. Mitochondr Physiol Network 24.02.
Komlodi 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. MitoFit Preprints 2021.2.v2. doi:10.26124/mitofit:2021-0002.v2
MitoFit pdf
Coupling and pathway control of coenzyme Q redox state and respiration in isolated mitochondria
2021-05-06
Huete-Ortega M, Di Marcello M, Iglesias-Gonzalez J, Gnaiger E (2020) High-resolution respirometry for chloroplast and mitochondrial bioenergetics in Chlamydomonas reinhardtii ― towards biotechnology exploitations. MitoFit Preprint Arch doi:10.26124/mitofit:ea20.algaeurope.0001.
MitoFit pdf
High-resolution respirometry for chloroplast and mitochondrial bioenergetics in Chlamydomonas reinhardtii ― towards biotechnology exploitations
2020-11-30
Huete-Ortega M, Di Marcello M, Doerrier C, Gnaiger E (2021) Substrate-uncoupler-inhibitor-titration protocols for photosynthesis in Chlamydomonas reinhardtii. MitoFit Preprints 2021 (in prep).
Doerrier C, Gnaiger E (2021) Simultaneous measurement of oxygen consumption and NADH redox states as a metabolic marker in pathophysiological conditions. (in prep)
Cardoso LHD, Doerrier C, Gnaiger E (2021) Magnesium Green for fluorometric measurement of ATP production does not interfere with mitochondrial respiration. MitoFit Preprints 2021.1. doi:10.26124/mitofit:2021-0001
MitoFit pdf
Magnesium Green for fluorometric measurement of ATP production does not interfere with mitochondrial respiration
2021-01-29


About NextGen-O2k

Top 10 reasons for Oroboros.jpg
Oroboros - as a driving force in mitochondrial physiology - extends the analytical and diagnostic power of high-resolution respirometry by integration of NADH- and Q-redox monitoring in the NextGen-O2k. We aim at establishing the Oroboros quality control management for dissemination to our worldwide O2k-Network laboratories. This will become an effective contribution to address 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 will support key developments in mitochondrial medicine. In addition, we expand our business to algal biotechnology and ecology with the photobiology module of the NextGen-O2k, widening our focus from medicine to environment and climate.
- a partner in the Oroboros Ecosystem

Aims

Respiration and photosynthesis
Form the O2k to the NextGen-O2k
Between 1992 and 1994, Oroboros Instruments released its core technology for high-resolution respirometry (HRR), the Oxygraph-2k or O2k, for real-time monitoring of oxygen consumption of mitochondria, tissues, and living cells. Dissolved oxygen is measured using high-resolution polarographic oxygen sensors. Its main advantages are the unique signal stability and resolution of oxygen. HRR is the core technology present in every single of the devices and it will remain the basis of the NextGen-O2k.
In 2012, the updated version of the O2k was launched, the O2k-FluoRespirometer. The integrated Smart Fluo-Sensors combine fluorescence excitation and detection of defined wavelengths that in combination with fluorophores allow for the measurement of additional mitochondrial parameters, such as membrane potential, H2O2 production, calcium concentration, and ATP production.
The unique software DatLab has been developed for data acquisition and analysis. DatLab allows the researcher to perform calibrations (including automatic barometric and temperature adjustments), background correction, experimental SUIT (substrate-uncoupler-inhibitor-titration) protocols and data analysis.


With the O2k core technology, developed exclusively by Oroboros for high-resolution measurement of oxygen flux, the NextGen-O2k was developed.


Oroboros products timeline.jpg
O2k
High-resolution respirometry (HRR) and unique combinations of the O2k with





NextGen-O2k modules.jpg
NextGen-O2k
Extend high-resolution respirometry (HRR) to the all-in-one device
The NextGen-O2k is a MultiSensor all-in-one instrument for in-depth studies of (1) the mitochondrial role in human diseases, and (2) mitochondria and chloroplasts ability in regulating algal growth and metabolite production rates.
(+2) Controlled wavelengths and light intensity: PhotoBiology-Module
  • Save on experimental time, reagents and samples
  • Minimize errors and ensure reproducible and accurate results
  • Highest oxygen sensitivity (5 nM detection limit)
  • Exceptional resolution (wide oxygen concentration range, between 0 - 1,000 μM)
  • Robustness (>10 years lifespan)



Objectives

The overall goal of this Phase 2 project is to complete the technical and scientific development of the new NextGen-O2k and prepare it for market launch.
WP1
» Technical developments
WP2
» Scientific developments
WP3
» Documentation
WP4
» Exploitation
WP5
» Dissemination
WP6
» Project management

Coordinator

Oroboros Instruments GmbH

Network

Partners

Our partners WGT-Elektronik and HTech have been essential in the development of the NextGen-O2k.
WGT Logo 2012.jpg
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Early testers

The influence of KOLs and Early testers/adopters will be used to build awareness of NextGen-O2k among researchers, academic and industrial partners. We have confirmed relevant mitochondrial and algal biotechnology researchers who want to collaborate as early testers:


Biomedical research Algal research
Prof. Massimo Zeviani a,b, Dr. Carlo Viscomi c, Prof. Mervyn Singer a

a Medical Research Council - Mitochondrial Biology Unit, University of Cambridge, UK
b Department of Neurosciences, University of Padova, IT
c Department of Biomedical Sciences, University of Padova, IT

Dr. Tomas Morosinotto

Department of Biology, University of Padova, IT

Prof. Pablo Garcia-Roves

Department of Physiological Sciences, Faculty of Medicine and Health Sciences, University of Barcelona and Bellvitge Biomedical Research Institute (IDIBELL), ES

Prof. Yagut Allahverdiyeva-Rinne

Faculty of Technology, University of Turku, FI

Dr. Anthony Molina

Division of Geriatrics and Gerontology, Department of Medicine, University of San Diego, US

Prof. Claire Remacle

Genetics and Physiology of Microalgae, InBios/Phytosystems Research Unit, University of Liege, BE

Dr. Marten Szibor

Clinic for Heart and Thorax Surgery, Jena University Hospital, DE

Prof. Howard Jacobs

Institute of Biotechnology, University of Helsinki, FI

Dr. Alexander Karabatsiakis

Institute of Psychology, Department of Clinical Psychology, University of Innsbruck, AT

NextGen-O2k events

 WhenWhere
NextGen-O2k Launch event 2021 Innsbruck AT2021-10-13
NextGenO2k logo.jpg
Innsbruck AT, 2021 Oct 13-19. NextGen-O2k Launch event, NextGen-O2k SME2.
World summit on Diabetes 2021 Virtual2021-06-16Vitual, 2021 Jun 16-17. World Summit on Diabetes: New technologies and practical approaches: Diabetes and Endocrine disorders
4th edition Metabolism & Cancer 2021 Virtual2021-05-27Virtual, 2021 May 27-29. 4th edition Metabolism & Cancer.
ISAP 2021 Virtual2021-05-14Virtual, 2021 May 14-Aug 13. 7th Conference of the International Society for Applied Phycology - ISAP2021.
Career Event - FH Campus 2021 Wien AT2021-01-26Virtual, 2021 Jan 26. Career Event of the FH Campus Wien.
Virtual OroDM022021
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Virtual Oroboros Distributor Meeting, OroDM02
AlgaEurope 2020 Virtual Event2020-12-01
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Rome, IT, 2020 Dec 1-5. AlgaEurope 2020.
Long Night of Research 2020 Virtual Event2020-10-09
LNF.JPG
Virtual Event, 2020 Oct 09. Oroboros at Long Night of Research, CCB. The diagnostic bioenergetic report – a milestone on the way to mitochondrial fitness and physical well-being.
36th Congress Czech Nutrition Society 2020 Hradec Kralove CZ2020-09-10Hradec Kralove, CZ, 2020 Sep 10-12 . 36th annual international congress of Czech Nutrition Society
MitoEAGLE WG2 Innsbruck AT2020-03-09
COST Action MitoEAGLE
Innsbruck AT, 2020 Mar 9-13. WG 2 on human skeletal muscle fibers retreat - COST Action MitoEAGLE.
MitoEAGLE WG4 Innsbruck AT2020-02-26
COST Action MitoEAGLE
Innsbruck AT, 2020 Feb 26-28. WG 4 blood cell workshop and retreat - COST Action MitoEAGLE.
MiPNet25.10 Distributor engagement Tokyo JP2020-02-04
NextGenO2k logo.jpg
Tokyo,JP, 2020 Feb 4-7. On-site O2k-Workshop on High-Resolution Respirometry and partner distribution engagement.
MiPNet25.06 IOC145 Innsbruck AT2020-01-27
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Innsbruck AT, 2020 Jan 27-29. O2k-Coaching Days - Basic IOC145.
MiPNet25.03 IOC144 Innsbruck AT2020-01-20
OROBOROS-solo.png
Innsbruck AT, 2020 Jan 20-22. O2k-Coaching Days - Basic IOC144.
1st Myocardial Function Symposium 2020 Graz AT2020-01-10Graz, AT, 2020 Jan 10-11. 1st Myocardial Function Symposium: “Targets in cardiometabolic disease”.
NextGen-O2k SME2 coaching Orily Pratt part 22019-12-03
NextGenO2k logo.jpg
Innsbruck AT, 2019 Dec 03-04 NextGen-O2k SME2 coaching Orily Pratt (part 2), NextGen-O2k SME2.
MiPNet24.15 IOC143 Haryana IN2019-11-15
O2k-Network
Haryana IN, 2019 Nov 15. Oroboros O2k-Workshop on high-resolution respirometry: Assessment of mitochondrial function in health and disease IOC143.
Neurocon 2019 Haryana IN2019-11-15
Neurocon
Haryana, IN. 2019 Nov 15-18, Neurocon 2019.
MiP2019/MitoEAGLE Belgrade RS2019-10-13
COST Action MitoEAGLE
MiPsociety
Belgrade RS, 13-16 Oct 2019. 14th Conference on Mitochondrial Physiology: Mitochondrial function: changes during life cycle and in noncommunicable diseases - COST MitoEAGLE perspectives and MitoEAGLE WG and MC Meeting.
MiPNet24.13 IOC142 Fukuoka JP2019-10-03
O2k-Network
ASMRM & J-mit 2019, Fukuoka JP, 2019 Oct 03 Oroboros Lunch Seminar at ASMRM & J-mit 2019 Fukuoka JP - Explore the Mito-World! (IOC142).
ASMRM & J-mit 2019 Fukuoka JP2019-10-03
ASMRM LOGO.JPG
Fukuoka, JP, 2019 Oct 03-05. Joint ASMRM and J-mit Conference
MiPNet24.02 IOC141 Schroecken AT2019-09-23
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Schroecken AT, 2019 Sep 23-28 Oroboros O2k-Workshop on high-resolution respirometry (HRR), IOC141.
NextGen-O2k SME2 coaching Samia Kappe part 12019-09-12
NextGenO2k logo.jpg
Innsbruck AT, 2019 Sep 10-11 NextGen-O2k SME2 coaching Samia Kappe (part 1), NextGen-O2k SME2.
FEBS Workshop Ageing 2019 Innsbruck AT2019-09-09
FEBS
Innsbruck, AT. 2019 Sep 09-12, FEBS Workshop “Ageing and Regeneration”.
Mitochondrial Physiology ‐ from Organelle to Organism 2019 Copenhagen DK2019-08-19
COST Action MitoEAGLE
Copenhagen DK , 2019 Aug 19-23. Mitochondrial Physiology ‐ from Organelle to Organism
NextGen-O2k SME2 coaching Orily Pratt part 12019-07-22
NextGenO2k logo.jpg
Innsbruck AT, 2019 Jul 22-23 NextGen-O2k SME2 coaching Orily Pratt (part 1), NextGen-O2k SME2.
MiPschool Coimbra 20192019-07-08
COST Action MitoEAGLE
MiPsociety
Coimbra, PT. 2019 Jul 08-11, 12th MiP/MitoEAGLE Training School 2019. Mitochondrial respiratory physiology: Challenges on data sharing, reproducibility, and interpretation.
OroDM01 Innsbruck AT2019-07-01
NextGenO2k logo.jpg
Innsbruck AT, 2019 Jul 01-03 Oroboros Distributor Meeting, OroDM01.
MiPNet24.04 IOC140 Amsterdam NL2019-06-23
O2k-Network
Amsterdam NL, 2019 Jun 23 Pre-conference Oroboros O2k-Workshop on high-resolution respirometry (HRR), IOC140.
MiPNet24.01 IOC139 Schroecken AT2019-06-17
O2k-Workshops.png
Schroecken AT, 2019 June 17-22 Oroboros O2k-Workshop on high-resolution respirometry (HRR), IOC139.


NextGen-O2k dissemination

NextGen-O2k press releases


Gentle Science

In addition to O2k-Workshops and on-site training, Oroboros will disseminate the not-for-profit initiatives on Mitochondrial Physiology and Bioenergetics Science Camps, specialized Mitochondrial Physiology Conferences, and Preprint publications:
» Oroboros is a key player in support of the COST Action MitoEAGLE
» MitoGlobal - the world-wide platform for societies and organizations supporting mitochondrial research and medicine
» MitoEAGLE with cooperation by Oroboros launched MitoFit Preprints - the preprint server for mitochondrial physiology and bioenergetics
» A vision on preprints
» Mitochondrial respiratory states and rates : >530 coauthors join forces towards harmonization on nomenclature
» Gentle Science - recognizes the responsibility of the scientific community - for the quality of science, the quality of life in science, and its mission


MitoEAGLE Q-workshop

MitoFit


Support

Template NextGen-O2k.jpg
Supported by project NextGen-O2k which has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No. 859770.

MitoPedia: NextGen-O2k

» NextGen-O2k«
TermAbbreviationDescription
Hydrogen peroxideH2O2
Hydrogen peroxide
Hydrogen peroxide, H2O2 or dihydrogen dioxide, is one of several reactive oxygen intermediates generally referred to as reactive oxygen species (ROS). It is formed in various enzyme-catalyzed reactions (e.g., superoxide dismutase) with the potential to damage cellular molecules and structures. H2O2 is dismutated by catalase to water and oxygen. H2O2 is produced as a signaling molecule in aerobic metabolism and passes membranes more easily compared to other ROS.
Light-enhanced dark respirationLEDRLight-enhanced dark respiration LEDR is a sharp (negative) maximum of dark respiration in plants in response to illumination, measured immediately after switching off the light. LEDR is supported by respiratory substrates produced during photosynthesis and closely reflects light-enhanced photorespiration (Xue et al 1996). Based on this assumption, the the total photosynthetic oxygen flux TP is calculated as the sum of the measured net photosynthetic oxygen flux NP plus the absolute value of LEDR.
Mitochondrial membrane potentialmtMP, Δψ [V]The mitochondrial membrane potential, mtMP, is the electric part of the protonmotive force, ΔpH+.

Δψ = ΔpH+ - ΔµH+ / F

mtMP or Δψ is the potential difference across the inner mitochondrial (mt) membrane, expressed in the electric unit of volt [V]. Electric force of the mitochondrial membrane potential is the electric energy change per ‘motive’ electron or per electron moved across the transmembrane potential difference, with the number of ‘motive’ electrons expressed in the unit coulomb [C].
NextGen-O2k
NextGen-O2k InstrumentNextGen-O2k Instrument
NextGen-O2k Technical developments
Oxygen kineticsOxygen kinetics describes the dependence of respiration of isolated mitochondria or cells on oxygen partial pressure. Frequently, a strictly hyperbolic kinetics is observed, with two parameters, the oxygen pressure at half-maximum flux, p50, and maximum flux, Jmax. The p50 is in the range of 0.2 to 0.8 kPa for cytochrome c oxidase, isolated mitochondria and small cells, strongly dependent on Jmax and coupling state.
PB-ModulePB-ModuleThe PB-Module has been developed for conducting measurements of PhotoBiology, including photosynthesis. It consists of the PB-Sensor and electronic components which are an integral part of the NextGen-O2k. Measurements are recorded and evaluated with the DatLab 8 software.
PB-SensorThe PB-Sensor has been designed as a part of the PB-Module to provide with an external source of light, in the range of the blue (451 nm) or red (634 nm) wavelengths. This enables experiments for evaluating the production/consumption of O2 in the presence of light. The PB-Sensor consists on one LED and one photodiode mounted on the sensor tip behind a plastic cover. The LED emits with blue or red light on the sample and its intensity can be regulated from 0 to 5000 µEinsteins*m-2*s-1. The photodiode provides real-time measurement of the light intensity allowing for continuous adjustment to the desired value, which can also help compensate for the temperature rise due to the operation of the LED.
PhotoBiologyPBPhotoBiology is the science of the effect of light on biological processes. This includes photosynthesis, photochemistry, photophysics, photomorphogenesis, vision, bioluminescence, circadian rhythms and photodynamic therapy. Phototoxicity results from non-ionizing radiation (i.e. ultraviolet, visible and infrared radiation). Non-ionizing radiation is any type of electromagnetic radiation that does not carry enough energy per quantum (photon energy below 10 eV) to completely remove an electron from an atom or molecule. When photons interact with molecules, the molecules can absorb the photon energy and become excited, reacting with surrounding molecules and stimulating "photochemical" and "photophysical" changes. Respiration may be affected by light during photosynthesis or in dark respiration, with the transient response of light-enhanced dark respiration.
Q-ModuleQ-ModuleThe Q-Module, developed both for measuring the Q redox state and for cyclic voltammetry measurements, is an integral part of the NextGen-O2k and consists of the Q-Sensor, integrated electronic components in the O2k, and the DatLab software.
Q-SensorThe Q-Sensor has been designed as a part of the Q-Module for measurements with cyclic voltammetry and voltammetry, allowing for analysis of the Q redox state. The Q-Stopper with the reference electrode is called Q-Sensor, which is plugged in the NextGen-O2k. A three-electrode system is used to detect the Q redox state. Two of the three electrodes (glassy carbon and platinum electrode) are built into the Q-Stopper, while the reference electrode is removable (Reference-Electrode\2.4 mm).
Template NextGen-O2k.jpg


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Bioblast links: Q - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>



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Bioblast links: NADH - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>

Mitochondrial NADH and respiration

» Nicotinamide adenine dinucleotide
» NADH electron transfer-pathway state
» N-junction
» Complex I
» Rotenone
» Amytal
» Piericidin

Mitochondrial NADH production

» Tricarboxylic acid cycle
» Fatty acid oxidation

NextGen-O2k

» NextGen-O2k




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Bioblast links: Hydrogen peroxide - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>



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Bioblast links: ATP production - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>

Phosphorylation pathway

» Phosphorylation pathway
» ATP synthase
» Adenine nucleotide translocase
» Phosphate carrier

Phosphorylation pathway substrates

» ADP
» ATP
» Inorganic phosphate

Phosphorylation pathway inhibitors

» Oligomycin
» Carboxyatractyloside
» Atractyloside
» Bongkrekik acid

Coupling control

» Oxidative phosphorylation
» OXPHOS capacity
» P-L control efficiency
» ROUTINE respiration
» R-L control efficiency

Respiratory complexes and coupling

» Complex I
» Complex III
» Complex IV
» Proton pump
» Electron-transfer-pathway state

ATP production measurement

» Magnesium Green



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Bioblast links: Calcium - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>



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Bioblast links: PhotoBiology and plant physiology - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>

PhotoBiology: photosynthesis

» PhotoBiology
» Photosynthesis

Plant physiology: respiration

» Photorespiration
» Chlororespiration
» Light-enhanced dark respiration

NextGen-O2k and PB-Module

» NextGen-O2k
» PB-Module
» PB-Sensor



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MitoPedia:NextGen-O2k