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| |journal=Mitochondr Physiol Network | | |journal=Mitochondr Physiol Network |
| |abstract=Fasching M, Sumbalova Z, Gnaiger E (2013) O2k-Fluorometry: HRR and H<sub>2</sub>O<sub>2</sub> production in mouse brain mitochondria. Mitochondr Physiol Network 17.17. | | |abstract=Fasching M, Sumbalova Z, Gnaiger E (2013) O2k-Fluorometry: HRR and H<sub>2</sub>O<sub>2</sub> production in mouse brain mitochondria. Mitochondr Physiol Network 17.17. |
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| [[High-resolution respirometry]] [1] is extended by MultiSensor techniques allowing the simultaneous measurement of oxygen consumption and one additional parameter (mitochondrial membrane potential, pH, Ca<sup>2+</sup>, or NO concentration [2-5]). Fluorometric methods are available for a wide range of analytical parameters of major interest in mitochochondrial physiology: H<sub>2</sub>O<sub>2</sub> production, mitochondrial membrane potential, intracellular pH, Ca<sup>2+</sup>, Mg<sup>2+</sup>, and NADH levels.
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| Advantages of the simultaneous measurement of additional parameters in a single respirometric chamber:
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| # Quality control of preparations by respirometric performance.
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| # Objective exclusion criteria for protocol related problems, e.g. overtitration of uncouplers, side effects of TPP<sup>+</sup> or fluorophores.
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| # Elimination of artifacts of normalization for a mitochondrial marker when combining two methods in a single chamber.
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| # Additional information is acquired for a limited amount of sample.
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| # Oxygen dependence of parameters can be studied.
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| Here we introduce the O2k-Fluorescence LED2-Module, describe one selected stage in its development, and show its application for the simultaneous measurement of respiration and H<sub>2</sub>O<sub>2</sub> production of mouse brain mitochondria.
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| :>> Product: [http://www.oroboros.at/?oxygraph OROBOROS O2k], [[O2k-Catalogue_OROBOROS| O2k-Catalogue]] | | :>> Product: [http://www.oroboros.at/?oxygraph OROBOROS O2k], [[O2k-Catalogue_OROBOROS| O2k-Catalogue]] |
| |mipnetlab=AT_Innsbruck_OROBOROS, AT Innsbruck MitoCom | | |mipnetlab=AT_Innsbruck_OROBOROS, AT Innsbruck MitoCom |
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| |additional=O2k-Demo, O2k-MultiSensor | | |additional=O2k-Demo, O2k-MultiSensor |
| }} | | }} |
| == Further Information ==
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| '''References'''
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| # [[O2k-Publications: O2k-Fluorometry]]
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| # [[Pesta 2012 Methods Mol Biol|Pesta D, Gnaiger E (2012) High-resolution respirometry. OXPHOS protocols for human cells and permeabilized fibres from small biopisies of human muscle. Methods Mol Biol 810: 25-58.]]
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| # [[Aguirre 2010 Biochim Biophys Acta|
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| Aguirre E, RodrΓguez-JuΓ‘rez 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: 557-565.]]
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| # [http://www.oroboros.at/index.php?mipnetanalyzer MiPNetAnalyzer]
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| # [[Hickey_2012_J_Comp_Physiol_B|Hickey AJ, Renshaw GM, Speers-Roesch B, Richards JG, Wang Y, Farrell AP, Brauner CJ (2012) A radical approach to beating hypoxia: depressed free radical release from heart fibres of the hypoxia-tolerant epaulette shark (Hemiscyllum ocellatum). J Comp Physiol B 182: 91-100.]]
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| # [[Krumschnabel_2014_Methods_Mol_Biol| Krumschnabel G, Fontana-Ayoub M, Sumbalova Z, Heidler J, Gauper K, Fasching M, Gnaiger E (2014) Simultaneous high-resolution measurement of mitochondrial respiration and hydrogen peroxide production. Methods Mol Biol [in press].]]
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| # [[Gnaiger 2012 MitoPathways|Gnaiger E (2012) Mitochondrial Pathways and Respiratory Control. An Introduction to OXPHOS Analysis. 3rd ed. Mitochondr Physiol Network 17.18. OROBOROS MiPNet Publications, Innsbruck: 64 pp.]]
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O2k-Fluorometry: HRR and H2O2 production in mouse brain mitochondria. Β»Bioblast pdfΒ«
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Β» Versions
OROBOROS (2014-11-14) Mitochondr Physiol Network
Abstract: Fasching M, Sumbalova Z, Gnaiger E (2013) O2k-Fluorometry: HRR and H2O2 production in mouse brain mitochondria. Mitochondr Physiol Network 17.17.
- >> Product: OROBOROS O2k, O2k-Catalogue
β’ O2k-Network Lab: AT_Innsbruck_OROBOROS, AT Innsbruck MitoCom
Labels: MiParea: Respiration, Instruments;methods
Organism: Mouse
Tissue;cell: Nervous system
Preparation: Isolated mitochondria
Regulation: Uncoupler
Coupling state: LEAK, OXPHOS, ETS"ETS" is not in the list (LEAK, ROUTINE, OXPHOS, ET) of allowed values for the "Coupling states" property.
HRR: Oxygraph-2k, O2k-Fluorometer, Protocol"Protocol" is not in the list (Oxygraph-2k, TIP2k, O2k-Fluorometer, pH, NO, TPP, Ca, O2k-Spectrophotometer, O2k-Manual, O2k-Protocol, ...) of allowed values for the "Instrument and method" property.
O2k-Demo, O2k-MultiSensor