NextGen-O2k

2022

NextGen-O2k

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

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 End: 2021-10-31
See: Accelerator data hub

NextGen-O2k publications, preprints, and abstracts

Link	Reference	Year	View
MiPNet26.12 NextGen-O2k: NADH-Module	NextGen-O2k: NADH-Module manual	2022-05-17	» Versions
MiPNet24.16 DatLab8.0: CV-Module	DatLab8.0: Cyclic voltammetry manual	2021-10-28	»Versions
MiPNet26.13 NextGen-O2k manual	NextGen-O2k manual	2022-07-25	» Versions
MiPNet24.12 NextGen-O2k: Q-Module	NextGen-O2k: Q-Module manual	2021-10-29	» Versions
MiPNet26.11 NextGen-O2k: PB-Module	NextGen-O2k: PB-Module manual	2021-10-29	» Versions
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. https://doi.org/10.26124/mitofit:2021-0005	2021	MitoFit Preprints 2021.05. Oxygen dependence of photosynthesis and light-enhanced dark respiration studied by High-Resolution PhotoRespirometry
Went 2021 Batchelor thesis	Went N (2021) Oxygen dependence of photosynthesis and light-enhanced dark respiration studied by High-Resolution PhotoRespirometry. Batchelor thesis:26 pp.	2021	Went 2021 Batchelor thesis
Vera-Vives 2022 BEC	Vera-Vives AM, Perin G, Morosinotto T (2022) High-resolution photosynthesis-irradiance curves in microalgae. Bioenerg Commun 2022.19. https://doi.org/10.26124/bec:2022-0019	2022	Bioenerg Commun 2022.19. published online 2022-12-15
Vera Vives 2022 MitoFit	Vera-Vives AM, Perin G, Morosinotto T (2022) The robustness of NextGen-O2k for building PI curves in microalgae. https://doi.org/10.26124/mitofit:2022-0019 — 2022-12-15 published in Bioenerg Commun 2022.19.	2022	MitoFit Preprints 2022.19. The robustness of NextGen-O2k for building PI curves in microalgae
Ravasz 2019 Abstract IOC141	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.
Pallag 2022 Int J Mol Sci	Pallag G, Nazarian S, Ravasz D, Bui D, Komlódi T, Doerrier C, Gnaiger E, Seyfried TN, Chinopoulos C (2022) Proline oxidation supports mitochondrial ATP production when Complex I is inhibited. https://doi.org/10.3390/ijms23095111	2022	Int J Mol Sci 23:5111. PMID: 35563503 Open Access
Pallag 2022 MitoFit Proline	Pallag G, Nazarian S, Ravasz D, Bui D, Komlódi T, Doerrier C, Gnaiger E, Seyfried TN, Chinopoulos C (2022) Proline oxidation leading to high electron flow through reduction of ubiquinone supports ATP production by F₁F_O-ATPase in mitochondria with inhibited Complex I. https://doi.org/10.26124/mitofit:2022-0001.v3 — 2022-05-04 published in https://doi.org/10.3390/ijms23095111	2022	MitoFit Preprints 2022.01.v3. Proline oxidation leading to high electron flow through reduction of ubiquinone supports ATP production by F₁F_O-ATPase in mitochondria with inhibited Complex I
Gnaiger 2022 Abstract Bioblast-PB	P03. Went N, Di Marcello M, Gnaiger Erich (2022) Oxygen dependence of photosynthesis and light-enhanced dark respiration studied by high-resolution PhotoRespirometry. Bioblast 2022: BEC Inaugural Conference. In: https://doi.org/10.26124/bec:2022-0001 »MitoFit Preprint«
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. https://doi.org/10.26124/bec:2021-0003	2021	Bioenerg Commun 2021.3. published online 2021-11-11
Huete-Ortega 2020 MitoFit Preprint Arch EA	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. https://doi.org/10.26124/mitofit:ea20.algaeurope.0001	2020	MitoFit Preprint Arch EA20.1. High-resolution respirometry for chloroplast and mitochondrial bioenergetics in Chlamydomonas reinhardtii ― towards biotechnology exploitations
Doerrier 2022 MitoFit NADH	Doerrier C, Cardoso LHD, Gnaiger E (2022) Simultaneous measurement of oxygen consumption and NAD(P)H/NAD(P)⁺-redox state in isolated mitochondria. (in prep)
Copsey 2021 Sci Rep	Copsey AC, Barsottini MRO, May B, Xu F, Albury MS, Young L, Moore AL (2021) Kinetic characterisation and inhibitor sensitivity of Candida albicans and Candida auris recombinant AOX expressed in a self-assembled proteoliposome system. Sci Rep 11:14748.	2021	PMID: 34285303 Open Access
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. https://doi.org/10.26124/bec:2021-0001	2021	Bioenerg Commun 2021.1. published online 2021-06-30
Cardoso 2022 Abstract Bioblast	4.3. «10+5» Cardoso Luiza HD, Doerrier C, Donnelly C, Komlódi T, Gnaiger E (2022) Redox monitoring and respiration - a new horizon with the NextGen-O2k. Bioblast 2022: BEC Inaugural Conference. In: https://doi.org/10.26124/bec:2022-0001

About NextGen-O2k

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

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.

O2k

High-resolution respirometry (HRR) and unique combinations of the O2k with

Fluorometry: the O2k-FluoRespirometer
Potentiometry: the O2k-pH ISE-Module and O2k-TPP+ ISE-Module
Amperometric measurement of nitric oxide: the O2k-NO Amp-Module

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.

(+1) Detection of the redox state of the Q-junction: MiPNet24.12 NextGen-O2k: Q-Module

(+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

WP4

WP5

WP6

Coordinator

Oroboros Instruments GmbH

Network

Partners

Our partners WGT-Elektronik and HTech have been essential in the development of the NextGen-O2k.

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

NextGen-O2k events

	When	Where
MiPNet28.02 IOC Schroecken AT	2023-10-02	Schroecken AT, 2023 Oct 02-07. Oroboros O2k-Workshop on high-resolution respirometry (HRR), IOC.
MiPNet28.01 IOC160 Schroecken AT	2023-06-19	Schroecken AT, 2023 Jun 19-24. Oroboros O2k-Workshop on high-resolution respirometry (HRR), IOC160.
MiPNet28.05 IOC158 Innsbruck AT	2023-02-27	Innsbruck AT, 2023 Feb 27 - Mar 01. O2k-Coaching Days - Basic, IOC158.
Virtual OroDM02	2023	Virtual Oroboros Distributor Meeting, OroDM02
MiPNet27.04 IOC155 Schroecken AT	2022-10-03	Schroecken AT, 2022 Oct 03-08. Oroboros O2k-Workshop on high-resolution respirometry (HRR), IOC155.
MiPNet27.05 Schroecken BEC tutorial-Living Communications pmP	2022-09-30	Schroecken AT, 2022 Sep 30-Oct 03. BEC tutorial-Living Communications: pmF to pmP — pre IOC155.
MiPNet27.03 IOC154 Innsbruck AT	2022-08-01	Innsbruck AT, 2022 Aug 01-05. O2k-Coaching Days - O2k-FluoRespirometer and NextGen-O2k, IOC154.
MiPNet27.02 IOC153 Innsbruck AT	2022-07-01	Innsbruck AT, 2022 Jul 01-02. NextGen-O2k All-in-one Demo Workshop IOC153.
MiPNet 26.16 NextGen-O2k Summit 2021 Virtual	2021-10-20	Virtual, 2021 Oct 20. NextGen-O2k Summit, NextGen-O2k SME2. MiPNet 26.16.
The 4th Academic Symposium of Metabolic Biology Branch 2021 Guangxi CN	2021-07-29	Guangxi Province , CN, 2021 Jul 29 - Aug 02. The 4th Academic Symposium of Metabolic Biology Branch of Chinese Biophysical Society.
19th Beijing Conference and Exhibition on Instrumental Analysis 2021 Beijing CN	2021-07-27	Beijing, CN, 2021 Sep 27-29. 19th Beijing Conference and Exhibition on Instrumental Analysis.
19th Chinese Biophysics congress 2021 Anhui CN	2021-07-23	Anhui Province , CN, 2021 Jul 23-26. 19th Chinese Biophysics congress.
MiPNet 26.15 NextGen-O2k Event Photobiology: Algal bioenergetics Innsbruck AT	2021-07-15	Innsbruck AT, 2021 Jul 15-16. NextGen-O2k Event Photobiology: Algal bioenergetics. MiPNet 26.15.
4th China Symposium on Nerve Control Metabolism 2021 Hangzhou city CN	2021-06-18	Hangzhou city, CN, 2021 Jun 18-19. The 4th China Symposium on Neuro-Controlled Metabolism.
World summit on Diabetes 2021 Virtual	2021-06-16	Vitual, 2021 Jun 16-17. World Summit on Diabetes: New technologies and practical approaches: Diabetes and Endocrine disorders
ESCI 2021 Virtual	2021-06-09	Virtual, 2021 Jun 9-11. 55^th Annual Scientific Meeting of the European Society for Clinical Investigation, ESCI 2021.
4th edition Metabolism & Cancer 2021 Virtual	2021-05-27	Virtual, 2021 May 27-29. 4th edition Metabolism & Cancer.
24th Kalorimetrietage 2021 Braunschweig DE	2021-05-26	Braunschweig, DE, 2021 May 26-28. 24th Kalorimetrietage
ISAP 2021 Virtual	2021-05-14	Virtual, 2021 May 14-Aug 13. 7th Conference of the International Society for Applied Phycology - ISAP2021.
Career Event - FH Campus 2021 Wien AT	2021-01-26	Virtual, 2021 Jan 26. Career Event of the FH Campus Wien.
AlgaEurope 2020 Virtual Event	2020-12-01	Rome, IT, 2020 Dec 1-5. AlgaEurope 2020.
2020 PaduaMuscleDays Padua IT	2020-11-19	Virtual, 2020 Nov 19-21. PaduaMuscleDays - 30 years of translational research
Analytica China 2020 Shanghai CN	2020-11-16	Shanghai, CN, 2020 Nov 16-18.Analytica China Exhibition - 10th International Trade Fair for Laboratory Technology, Analysis, Biotechnology and Diagnostics.
Long Night of Research 2020 Virtual Event	2020-10-09	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.
12th ÖGMBT Annual Meeting 2020 Virtual Event	2020-09-21	Virtual Event, 2020 Sep 21-23. 12th ÖGMBT Annual Meeting - Biomolecules in/for 21st century
MitoEAGLE WG2 Innsbruck AT	2020-03-09	Innsbruck AT, 2020 Mar 9-13. WG 2 on human skeletal muscle fibers retreat - COST Action MitoEAGLE.
MitoEAGLE WG4 Innsbruck AT	2020-02-26	Innsbruck AT, 2020 Feb 26-28. WG 4 blood cell workshop and retreat - COST Action MitoEAGLE.
MiPNet25.10 Distributor engagement Tokyo JP	2020-02-04	Tokyo,JP, 2020 Feb 4-7. On-site O2k-Workshop on High-Resolution Respirometry and partner distribution engagement.
MiPNet25.06 IOC145 Innsbruck AT	2020-01-27	Innsbruck AT, 2020 Jan 27-29. O2k-Coaching Days - Basic IOC145.
MiPNet25.03 IOC144 Innsbruck AT	2020-01-20	Innsbruck AT, 2020 Jan 20-22. O2k-Coaching Days - Basic IOC144.
1st Myocardial Function Symposium 2020 Graz AT	2020-01-10	Graz, AT, 2020 Jan 10-11. 1st Myocardial Function Symposium: “Targets in cardiometabolic disease”.
NextGen-O2k SME2 coaching Orily Pratt part 2	2019-12-03	Innsbruck AT, 2019 Dec 03-04 NextGen-O2k SME2 coaching Orily Pratt (part 2), NextGen-O2k SME2.
MiPNet24.15 IOC143 Haryana IN	2019-11-15	Haryana IN, 2019 Nov 15. Oroboros O2k-Workshop on high-resolution respirometry: Assessment of mitochondrial function in health and disease IOC143.
Neurocon 2019 Haryana IN	2019-11-15	Haryana, IN. 2019 Nov 15-18, Neurocon 2019.
MiP2019/MitoEAGLE Belgrade RS	2019-10-13	Belgrade RS, 13-16 Oct 2019. 14^th 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 JP	2019-10-03	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 JP	2019-10-03	Fukuoka, JP, 2019 Oct 03-05. Joint ASMRM and J-mit Conference
MiPNet24.02 IOC141 Schroecken AT	2019-09-23	Schroecken AT, 2019 Sep 23-28 Oroboros O2k-Workshop on high-resolution respirometry (HRR), IOC141.
NextGen-O2k SME2 coaching Samia Kappe part 1	2019-09-12	Innsbruck AT, 2019 Sep 10-11 NextGen-O2k SME2 coaching Samia Kappe (part 1), NextGen-O2k SME2.
FEBS Workshop Ageing 2019 Innsbruck AT	2019-09-09	Innsbruck, AT. 2019 Sep 09-12, FEBS Workshop “Ageing and Regeneration”.
Mitochondrial Physiology ‐ from Organelle to Organism 2019 Copenhagen DK	2019-08-19	Copenhagen DK , 2019 Aug 19-23. Mitochondrial Physiology ‐ from Organelle to Organism
NextGen-O2k SME2 coaching Orily Pratt part 1	2019-07-22	Innsbruck AT, 2019 Jul 22-23 NextGen-O2k SME2 coaching Orily Pratt (part 1), NextGen-O2k SME2.
MiPschool Coimbra 2019	2019-07-08	Coimbra, PT. 2019 Jul 08-11, 12^th MiP/MitoEAGLE Training School 2019. Mitochondrial respiratory physiology: Challenges on data sharing, reproducibility, and interpretation.
OroDM01 Innsbruck AT	2019-07-01	Innsbruck AT, 2019 Jul 01-03 Oroboros Distributor Meeting, OroDM01.
MiPNet24.04 IOC140 Amsterdam NL	2019-06-23	Amsterdam NL, 2019 Jun 23 Pre-conference Oroboros O2k-Workshop on high-resolution respirometry (HRR), IOC140.
MiPNet24.01 IOC139 Schroecken AT	2019-06-17	Schroecken AT, 2019 June 17-22 Oroboros O2k-Workshop on high-resolution respirometry (HRR), IOC139.

NextGen-O2k dissemination

Cluster Award 2019

The NextGen-O2k was awarded as the best Tyrolean innovation of the year 2019 with the Cluster Award in the category Life Science. The Cluster Award is funded by the Standortagentur Tirol.

NextGen-O2k press releases

NextGen-O2k ermöglicht noch präzisere Echtzeitmessung der Photosynthese in Algen - WKO online, 2022-08-28
World Health Day: Discover how early diagnoses, virtual platforms and AI are improving our wellness - EIC community stories - online, 2022-04-07
Technical leap in mitochondrial research and applications - CORDIS EU research results - online, 2022-03-21
Tiroler Innovationspreis - Nominierter Technische Innovation - WKO Tirol, 2022-03-02
Tiroler Innovationspreis geht in die nächste Runde, Tiroler Wirtschaft online, 2022-02-11
Depression als Art 'Burnout' in den Zellen - Tiroler Tageszeitung online, Sep 2020, 2020-09-24
Rekord bei Patenten, Tirol auf Rang sechs - Tiroler Tageszeitung, July 2020, 2020-07-03
Oroboros Instrument revolutioniert Zellbiologische Wissenschaft - www.wirtschaftszeit.at; 2020-03-30
Beste Innovationen aus Tirol] www.economyaustria.at, 2019-12-16
Clusterawards 2019: Ausgezeichnete Innovationen aus Tirol science.apa.at, 2019-12-13
Tiroler Unternehmen für Innovationen mit Clusterawards 2019 ausgezeichnet - www.wirtschaftszeit.at; 2019-12-16
Clusterawards 2019: Ausgezeichnete Innovationen aus Tirol www.meinbezirk.at, 2019-12-13
Tiroler Forschungsunternehmen startet durch - Tiroler Landeszeitung, September 2019, S30-31, 2019-09-23
Innovationsförderung Tirol - Oroboros erhält Zuschuss in Millionenhöhe, Der Report 2019, Sonderheft der Wirtschaftsnachrichten, S12-13, 2019-09-18
Dem Leben beim Atmen zusehen Der Standard, Beilage Forschung spezial, S 13, 2019-08-07.
Turbo für Mitochondrienforschung und Oroboros Instruments, Standortagentur Tirol online, 2019-07-29
Tiroler Unternehmen Oroboros erhält 1.7 Millionen Euro EU-Förderung, tt.com, 2019-07-29
Turbo für Mitochondrienforschung udn Oroboros Instruments science.apa.at, online, 2019-07-29
1.7 Millionen Euro Förderung für Oroboros Instruments www.meinbezirk.at online, 2019-07-29
1,7 Millionen Euro EU-Flörderung für Tiroler KMU www.derbrutkasten.com online, 2019-07-29
Tiroler Erfindung macht in Europa Furore theworldnews.net online, 2019-07-29
Tiroler Erfindung macht in Europa Furore www.krone.at online, 2019-07-29
EUR 1,7 Mio Förderung für Tiroler KMU Oroboros Tiroler Tageszeitung, S 16, 2019-07-30
Tiroler Millionen-Erfindung Tiroler Krone, S 22, 2019-07-30

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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

Electron supply to the Q-junction: assessment of mitochondrial respiration, H2O2 flux and the redox state of the Q-pool. COST Action MitoEAGLE, Innsbruck, Nov 2018.

Support

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«

Term	Abbreviation	Description
Hydrogen peroxide	H₂O₂	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 respiration	LEDR	Light-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 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 potential	mtMP, ΔΨ_p⁺, Δ_elF_ep⁺ [V]	The mitochondrial membrane potential difference, mtMP or ΔΨ_p⁺ = Δ_elF_ep⁺, is the electric part of the protonmotive force, Δp = Δ_mF_eH⁺. Δ_elF_ep⁺ = Δ_mF_eH⁺ - Δ_dF_eH⁺ ΔΨ_p⁺ = Δp - Δµ_H+·(z_H⁺·F)^-1 ΔΨ_p⁺ is the potential difference across the mitochondrial inner membrane (mtIM), expressed in the electric unit of volt [V]. Electric force of the mitochondrial membrane potential is the electric energy change per ‘motive’ charge or per charge moved across the transmembrane potential difference, with the number of ‘motive’ charges expressed in the unit coulomb [C].
NADH-Module		The NADH-Module, is a component of the NextGen-O2k for simultaneous measurement of oxygen consumption and NAD(P)H autofluorescence. NAD(P)H autofluorescence is used to evaluate the redox state of the NAD(P)H-pool. The NADH-Module incorporates an UV light and NADH-Sensors which include a photodiode and specific filters.
NADH-Sensor		The NADH-Sensor has been developed as a part of the NADH-Module for simultaneous monitoring of oxygen consumption and NADH redox state. The NADH-Sensor is composed of a photodiode and equipped with three supergel R370 Italian blue filters (Rosco, US).
NextGen-O2k
NextGen-O2k Technical developments
Oxygen kinetics		Oxygen 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 J_max and coupling state.
PB Light Source		The PhotoBiology Light Source (PBLS) has been designed as a part of the PB-Module to provide with an external source of light. This enables experiments for evaluating the production of O₂ in the presence of light. The PBLS consists of one LED and one photodiode mounted on the PBLS head protected by a PMMA plastic cover. Three pairs of PBLS (white, blue, and red) are provided with the PB-Module. The light intensity can be regulated from 0 to 2750 µmol·s^-1·m^-2 (red PBLS), from 0 to 3000 µmol·s^-1·m^-2 (blue PBLS), from 0 to 3500 µmol·s^-1·m^-2 (white PBLS). An integrated photodiode provides real-time measurement of the light intensity allowing for continuous adjustment to the desired value.
PB-Module		The PB-Module has been developed for conducting measurements of PhotoBiology, including photosynthesis. It consists of the PB Light Source and electronic components which are an integral part of the NextGen-O2k. Measurements are recorded and evaluated with the DatLab 8 software.
PhotoBiology	PB	PhotoBiology 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.
Photosynthesis	PS	Photosynthesis is the process that converts light energy into chemical energy which is subsequently transformed to the physiological energy demand. Photosynthesis has a light-dependent and light-independent (dark) phase. In plants, algae, and cynobacteria, light energy is absorbed during the light phase by the pigment chlorophyll and used to split water and generate adenosine triphosphate (ATP) and reducing power - nicotinamide adenine dinucleotide phosphate (NADPH), with the net production of O₂ as a waste product. During the dark phase ATP and NADPH are used to synthesize carbohydrates from CO₂ through the metabolic pathway called Calvin-Benson cycle. Oxygenic photosynthesis is responsible for producing and maintaining the oxygen concentration of the Earth’s atmosphere. In bacteria such as cyanobacteria, photosynthesis involves the plasma membrane and the cytoplasm. In eukaryotic cells (plants and algae), photosynthesis takes place in the chloroplasts.
Q-Module	Q-Module	The Q-Module, developed for measuring the Q redox state and cyclic voltammetry, is supported by the NextGen-O2k and consists of the Q-Sensor, integrated electronic components in the O2k, and the DatLab software.
Q-Sensor		The 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).