MiPNet17.17 Amplex-Mouse-brain: Difference between revisions
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|year=2012 | |year=2012 | ||
|journal=Mitochondr Physiol Network | |journal=Mitochondr Physiol Network | ||
|abstract=High-resolution respirometry [1] is extended by MultiSensor techniques allowing the simultaneous measurement of oxygen consumption and one additional parameter (mitochondrial membrane potential, pH, | |abstract=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, 3]). Fluorometric methods are available for a wide range of analytical parameters of major interest in mitochochondrial physiology: H2O2 production, mitochondrial membrane potential, intracellular pH, Ca<sup>2+</sup>, Mg<sup>2+</sup>, and NADH levels. | ||
Advantages of the simultaneous measurement of additional parameters in a single respirometric chamber: | Advantages of the simultaneous measurement of additional parameters in a single respirometric chamber: | ||
# Quality control of preparations by respirometric performance. | # 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. | # Objective exclusion criteria for protocol related problems, e.g. overtitration of uncouplers, side effects of TPP<sup>+</sup> or fluorophores. | ||
# Elimination of artifacts of normalization for a mitochondrial marker when combining two methods in a single chamber. | # 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. | # Additional information is acquired for a limited amount of sample. | ||
# Oxygen dependence of parameters can be studied. | # 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 | 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. | ||
|keywords=Oxygraph-2k, High-resolution respirometry, Spectrofluorometry, MitoCom | |keywords=Oxygraph-2k, High-resolution respirometry, Spectrofluorometry, MitoCom | ||
|mipnetlab=AT_Innsbruck_OROBOROS | |mipnetlab=AT_Innsbruck_OROBOROS | ||
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|articletype=Workshop, MiPNet-online Publication | |articletype=Workshop, MiPNet-online Publication | ||
}} | }} | ||
# [[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.]] | |||
# [[Aguirre 2010 Biochim Biophys Acta| | |||
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.]] | |||
# [http://www.oroboros.at/index.php?mipnetanalyzer] |
Revision as of 16:48, 10 January 2013
Fasching M (2012) O2k-Fluorometry - A MitoCom Project. Mitochondr Physiol Network 17.17. |
Β» MiPNet17.17
Fasching M, Sumbalova Z, Gnaiger E (2012) Mitochondr Physiol Network
Abstract: 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, 3]). 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. β’ Keywords: Oxygraph-2k, High-resolution respirometry, Spectrofluorometry, MitoCom
β’ O2k-Network Lab: AT_Innsbruck_OROBOROS
Labels:
HRR: Oxygraph-2k, Spectrofluorimetry"Spectrofluorimetry" 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., MiPNet-Publication"MiPNet-Publication" 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.
- 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.
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