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High-resolution respirometry

From Bioblast


high-resolution terminology - matching measurements at high-resolution


High-resolution respirometry

Description

O2k-Fluorometer Series G.jpg

Mitochondrial function and dysfunction have gained increased interest over the past ten years, reflecting growing awareness of the fact that mitochondria play a pivotal role in human health and disease. The most appropriate way to analyze mitochondrial function is by means of O2k high-resolution respirometry (HRR) applying Substrate-uncoupler-inhibitor titration (SUIT) protocols that allow the diagnosis of multiple mitochondrial pathway defects in a single respirometry assay. Technically, HRR is based on the OROBOROS O2k, combining optimized chamber design, application of oxygen-tight materials, electrochemical sensors, Peltier-temperature control and software features (DatLab) to obtain the unique sensitive and quantitative resolution of oxygen concentration and oxygen flux, with a closed-chamber or open-chamber mode of operation (TIP2k). Standardized 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. The most important extension to O2k-MultiSensor systems is the O2k-Fluorometer.

Abbreviation: HRR

Reference: Gnaiger 1995 J Bioenerg Biomembr, Gnaiger 2008 POS, Gnaiger 2001 Respir Physiol


MitoPedia methods: Respirometry, Fluorometry, Spectrophotometry 


MitoPedia O2k and high-resolution respirometry: DatLab 

Dioxygen


What is an Oxygraph?

  • O2k: The O2k measures oxygen (dioxygen, O2) as a gas dissolved in aqueous solution.
  • With the O2k, experimental data are displayed not only of oxygen concentration over time, but real-time transformation of these data allows the experimenter to follow respiration (oxygen consumption, oxygen flux) of biological and biochemical samples while the experiment is still running.
  • An Oxygraph is not used for measurement of oxygen concentration in a tissue, but for high-resolution analysis of respiratory activity of biological or biochemical samples transferred into the experimental chamber.
  • Multiple O2k-units are offered as a ‘Power-O2k System’ for HRR combined with high-throughput - at high output. More details? Consult Power-O2k.


MitoPathways
Pesta and Gnaiger 2012


Specific substrate-uncoupler-inhibitor titration (SUIT) protocols are applied in studies of cell respiration or mitochondrial activity. In experiments with intact cells different coupling states can be studied to evaluate ROUTINE respiratory activity, respiration coupled to ATP production, respiratory excess capacity, residual oxygen consumption, and, by extension of conventional protocols, intactness of cells. In mitochondrial preparations such as isolated mitochondria, permeablized cells and tissue samples, or homogenized tissue, quantitative information is obtained on selected mitochondrial 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-universe.jpg
  • O2k: The O2k is the 8th 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 O2k 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-output.



OROBOROS INSTRUMENTS

OROBOROS INSTRUMENTS distributes the world-wide leading O2k-technology of high-resolution respirometry - O2k and HRR. The OROBOROS Company is fully scientifically oriented, with emphasis on continuous innovation. The extension of the Oxygraph-2k (O2k) to the O2k-Fluorometer has set a new standard for O2k-MultiSensor approaches (see OROBOROS O2k-Catalogue). The O2k is established internationally, with »4685 O2k-Publications in the scientific literature covering areas ranging from fundamental bioenergetics to the analysis of mitochondrial and metabolic diseases, advancing the rapidly growing field of preventive mitochondrial medicine. The OROBOROS science team actively participates in science and research publications by the OROBOROS science team and the OROBOROS lab continuously hosts international researchers as visiting scientists. OROBOROS has organized more than 100 international O2k-Workshops. The O2k-Network includes more than 760 Reference Laboratories in 49 countries.


MitoPedia: O2k and high-resolution respirometry

» MitoPedia: O2k hardware
» MitoPedia: DatLab
» MitoPedia: SUIT


Links

» MitoPedia: Respiratory states OXPHOS ROUTINE ETS LEAK - ROX