Talk:MiPNet24.10 H2O2 flux analysis
High-resolution respirometry is extended by O2k-MultiSensor modules allowing the simultaneous measurement of oxygen consumption and one additional parameter. 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.
In the publication by Krumschnabel et al. 2015 we introduce the O2k-Fluo 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. The paper by Makrecka-Kuka et al. 2015 focuses on the application of the Amplex red assay for determining H2O2 production in three different models of mitochondrial preparations.