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MiPNet17.17 Amplex-Mouse-brain

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Publications in the MiPMap
Fasching M (2012) O2k-Fluorometry - A MitoCom Project. Mitochondr Physiol Network 17.17.

» MiPNet17.17 Open Access

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:

  1. Quality control of preparations by respirometric performance.
  2. Objective exclusion criteria for protocol related problems, e.g. overtitration of uncouplers, side effects of TPP+ or fluorophores.
  3. Elimination of artifacts of normalization for a mitochondrial marker when combining two methods in a single chamber.
  4. Additional information is acquired for a limited amount of sample.
  5. 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. Keywords: Oxygraph-2k, High-resolution respirometry, Spectrofluorometry, MitoCom

O2k-Network Lab: AT_Innsbruck_OROBOROS


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


References

  1. 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.
  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