Difference between revisions of "Fasching 2012 Abstract Bioblast"

Revision as of 14:14, 12 November 2012

*Fasching M (2012) O2k-Fluorimetry - a MitoCom* project. Mitochondr Physiol Network 17.12.**

Link: MiPNet17.12 Bioblast 2012 - Open Access

Fasching M, Sumbalova Z, Gnaiger E (2012)

Event: Bioblast 2012

High-resolution respiromety [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]). Combining optical measurements with high-resolution respirometry further increased the range of analytical opportunities. In particular, 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.

The simultaneous measurement of additional parameters in a single respirometric chamber under strictly identical conditions offers important advantages: (i) respirometric performance provides a quality control of cells or mitochondrial preparations; (ii) overtitration of uncouplers, incomplete action of inhibitors, or non-saturating substrate concentrations are evaluated by the simultaneous response of multiple parameters, providing objective exclusion criteria; (iii) side effects of TPP⁺ or fluorophores (inhibition, dyscoupling) are detected by respiration and artifacts are, therefore, excluded; (iv) the direct relationship between the different parameters eliminates artifacts of normalization for a mitochondrial marker; (v) additional information is acquired for a limited amount of sample. In addition, there are practical and economical advantages, saving handling time and money. The glass chamber of the OROBOROS Oxygraph-2k (O2k) respirometer allows transmitting optical signals through the chamber wall. This extends MultiSensor applications, obtaining the optical signal in addition to the signals of the oxygen sensor and of other electrodes inserted through the stopper (e.g. respiration, mitochondrial membrane potential and H₂O₂ production).

[1] Gnaiger E (2008) Polarographic oxygen sensors, the oxygraph and high-resolution respirometry to assess mitochondrial function. In: Mitochondrial Dysfunction in Drug-Induced Toxicity (Dykens JA, Will Y, eds.) John Wiley: 327-352.

[2] Aguirre E, Rodríguez-Juárez 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.

[3] Mipnetanalyzer

• O2k-Network Lab: AT Innsbruck OROBOROS, AT Innsbruck Gnaiger E

Labels:

Organism: Mouse Tissue;cell: Neurons; Brain"Neurons; Brain" is not in the list (Heart, Skeletal muscle, Nervous system, Liver, Kidney, Lung;gill, Islet cell;pancreas;thymus, Endothelial;epithelial;mesothelial cell, Blood cells, Fat, ...) of allowed values for the "Tissue and cell" property.

HRR: Oxygraph-2k, Spectrophotometry"Spectrophotometry" 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.

Affiliations and author contributions

Mario Fasching (1), Zuzana Sumbalova (2), Erich Gnaiger (1,3)

(1) Oroboros Instruments GmbH, Innsbruck, Austria

(2) Comenius University, Bratislava, Slovak Republic

(3) Innsbruck Medical University, Austria. – [email protected]

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@@ Line 6: / Line 6: @@
 |event=[[Bioblast 2012]]
 |abstract=[[File:Mario.jpg|right|150px|Fasching Mario]] High-resolution respiromety [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]). Combining optical measurements with high-resolution respirometry further increased the range of analytical opportunities. In particular, 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.
+The simultaneous measurement of additional parameters in a single respirometric chamber under strictly identical conditions offers important advantages: (i) respirometric performance provides a quality control of cells or mitochondrial preparations; (ii) overtitration of uncouplers, incomplete action of inhibitors, or non-saturating substrate concentrations are evaluated by the simultaneous response of multiple parameters, providing objective exclusion criteria; (iii) side effects of TPP<sup>+</sup> or fluorophores (inhibition, dyscoupling) are detected by respiration and artifacts are, therefore, excluded; (iv) the direct relationship between the different parameters eliminates artifacts of normalization for a mitochondrial marker; (v) additional information is acquired for a limited amount of sample. In addition, there are practical and economical advantages, saving handling time and money.
+The glass chamber of the OROBOROS Oxygraph-2k (O2k) respirometer allows transmitting optical signals through the chamber wall. This extends MultiSensor applications, obtaining the optical signal in addition to the signals of the oxygen sensor and of other electrodes inserted through the stopper (e.g. respiration, mitochondrial membrane potential and H<sub>2</sub>O<sub>2</sub> production).
 [1] Gnaiger E (2008) Polarographic oxygen sensors, the oxygraph and high-resolution respirometry to assess mitochondrial function. In: Mitochondrial Dysfunction in Drug-Induced Toxicity (Dykens JA, Will Y, eds.) John Wiley: 327-352.
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 [3] [http://www.oroboros.at/index.php?mipnetanalyzer Mipnetanalyzer]
 |mipnetlab=AT Innsbruck OROBOROS, AT Innsbruck Gnaiger E,
 |journal=Mitochondr Physiol Network
 |articletype=Abstract