- high-resolution terminology - matching measurements at high-resolution
High-resolution respirometry
Description
High-resolution respirometry (HRR) is based on the OROBOROS Oxygraph-2k, combining chamber design, application of oxygen-tight materials, electrochemical sensors and electronics, Peltier-temperature control and software features (DatLab) to obtain a unique level of quantitative resolution of oxygen concentration and oxygen flux, with a closed-chamber or open-chamber mode of operation (TIP2k). Standardized two-point calibration of the polarographic oxygen sensor (static sensor calibration), calibration of the sensor response time (dynamic sensor calibration), and evaluation of instrumental background oxygen flux (systemic flux compensation) provide the experimental basis for high accuracy of quantitative results and quality control in HRR.
Abbreviation: HRR
Reference: Gnaiger_2008_POS, Gnaiger_2001_Respir Physiol
MitoPedia methods:
Respirometry,
Fluorometry,
Spectrophotometry
What is an Oxygraph?
- O2k: The Oxygraph-2k measures oxygen (dioxygen, O2) as a gas dissolved in aqueous solution.
- With the Oxygraph-2k, a graph is displayed not only of oxygen concentration over time, but of respiration (oxygen consumption, oxygen flux) of the biological sample.
An Oxygraph is not used for measurement of oxygen concentration in a tissue, but for high-resolution analysis of respiratory activity of a biological sample transferred into the experimental chamber.
Specific substrate-uncoupler-inhibitor titration (SUIT) protocols are applied in studies of cell respiration or mitochondrial activity. In such experiments, quantitative information is obtained on selected biochemical pathways, particularly of the process of oxidative phosphorylation (OXPHOS) in mitochondria and bacteria. Comprehensive OXPHOS analysis leads us beyond the highly informative process of oxygen consumption, when high-resolution respirometry is combined with O2k-MultiSensor Modules to measure simultaneously the production of reactive oxygen species or mitochondrial membrane potential, nitric oxide production, acid-base balance or calcium with O2k-Fluorometry or ion selective electrodes. In the O2k, these bioenergetic parameters are always evaluated in combination with the measurement of oxygen concentration and respiration.
- O2k: The Oxygraph-2k is the 2nd generation of our high-resolution respirometers. The first model was available since 1992. Development of the new O2k started in 2k (2000) and the first O2k was delivered in 2002.
- O2k: The Oxygraph-2k has two experimental chambers (k: Kammern, German for chambers). Why 2 ml?
- Power-O2k: Several O2k units are operated simultaneously (P1, P2, ..) to combine high-resolution respirometry with high-ouput.
Links
» Bioblast Glossary: Respiratory states
