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| | {{OROBOROS header page name}} |
| {{Publication | | {{Publication |
| |title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] O2k-Fluorometry: HRR and H<sub>2</sub>O<sub>2</sub> production in mouse brain mitochondria. | | |title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] O2k-Fluorometry: HRR and H<sub>2</sub>O<sub>2</sub> production in mouse brain mitochondria. |
| |info=[[Media:MiPNet17.17_Amplex-Mouse-brain.pdf|'''Open Access-MiPNet17.17.pdf''']] ;[http://www.bioblast.at/index.php/File:MiPNet17.17_Amplex-Mouse-brain.pdf Versions] | | |info=[[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/e/e2/MiPNet17.17_Amplex-Mouse-brain.pdf|Bioblast pdf]] Β»[http://www.bioblast.at/index.php/File:MiPNet17.17_Amplex-Mouse-brain.pdf Versions] |
| |authors=OROBOROS | | |authors=Oroboros |
| |year=2013-10 | | |year=2014-11-14 |
| |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. |
| | {{MiPNet pdf page linking to MitoPedia}} |
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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.
| | :Β» Product:[[O2k-Fluo_LED2-Module | O2k-Fluo LED2-Module]] |
| Advantages of the simultaneous measurement of additional parameters in a single respirometric chamber:
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| # Quality control of preparations by respirometric performance.
| | |mipnetlab=AT_Innsbruck_Oroboros, AT Innsbruck MitoCom |
| # 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 Oxygraph-2k], [[O2k-Catalogue_OROBOROS| O2k-Catalogue]]
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| |mipnetlab=AT_Innsbruck_OROBOROS, AT Innsbruck MitoCom | |
| }}
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| {{Labeling
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| |area=Respiration, Instruments;methods
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| |organism=Mouse
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| |tissues=Nervous system
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| |preparations=Isolated mitochondria
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| |topics=Uncoupler
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| |couplingstates=LEAK, OXPHOS, ETS
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| |substratestates=CI, CII, CIII, ROX
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| |instruments=Oxygraph-2k, O2k-Fluorometer, Protocol
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| |additional=O2k-Demo, O2k-MultiSensor
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| }} | | }} |
| == 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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