MiPNet17.17 Amplex-Mouse-brain: Difference between revisions
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# [http://www.oroboros.at/index.php?mipnetanalyzer MiPNetAnalyzer] | # [http://www.oroboros.at/index.php?mipnetanalyzer MiPNetAnalyzer] | ||
# [[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.]] | # [[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.]] | ||
# Krumschnabel G, | # [[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].]] | ||
# [[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.]] | # [[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.]] |
Revision as of 15:01, 3 October 2014
O2k-Fluorometry: HRR and H2O2 production in mouse brain mitochondria. |
Β» Open Access-MiPNet17.17.pdf ;Versions
OROBOROS (2013-10) 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.
High-resolution respirometry [1] is extended by MultiSensor techniques allowing the simultaneous measurement of oxygen consumption and one additional parameter (mitochondrial membrane potential, pH, Ca2+, or NO concentration [2-5]). Fluorometric methods are available for a wide range of analytical parameters of major interest in mitochochondrial physiology: H2O2 production, mitochondrial membrane potential, intracellular pH, Ca2+, Mg2+, and NADH levels. Advantages of the simultaneous measurement of additional parameters in a single respirometric chamber:
- Quality control of preparations by respirometric performance.
- Objective exclusion criteria for protocol related problems, e.g. overtitration of uncouplers, side effects of TPP+ or fluorophores.
- Elimination of artifacts of normalization for a mitochondrial marker when combining two methods in a single chamber.
- Additional information is acquired for a limited amount of sample.
- Oxygen dependence of parameters can be studied.
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 H2O2 production of mouse brain mitochondria.
- >> Product: OROBOROS Oxygraph-2k, 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"Isolated Mitochondria" is not in the list (Intact organism, Intact organ, Permeabilized cells, Permeabilized tissue, Homogenate, Isolated mitochondria, SMP, Chloroplasts, Enzyme, Oxidase;biochemical oxidation, ...) of allowed values for the "Preparation" property.
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, Fluorometry"Fluorometry" 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., 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
Further Information
References
- O2k-Publications: O2k-Fluorometry
- 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.
- 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.
- MiPNetAnalyzer
- 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.
- 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].
- 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.