Respirometry - OROBOROS Oxygraph-2k
OROBOROS Oxygraph-2k (O2k) is currently unavailable. For the alternative use, please see Sea horse XF96 & XF24-3 Analyzer at Bosch Molecular Biology Facility
- Diagnosis of acquired and genetic mitochondrial diseases
- Diagnosis of myopathies and neuromuscular pathologies
- Studies on cell function and cell death-apoptosis and necrosis, especially in non-mammalian cells such as yeast
Oxygraph-2k covers major areas of interest and demands:
- Routine analysis of oxidative phosphorylation (OXPHOS) in diverse fields of research such as redox signaling, oxidative stress, ischemia-reperfusion, aging, degenerative and inherited diseases, apoptosis, cancer research and drug testing, cell signaling and environmental stress.
- Full software support by DatLab includes automatic oxygen calibration, continuous display of oxygen concentration and respiratory flux, and on-line tabulation of final results during selected experimental sections. Standardized O2k quality controls eliminate interferences by instrumental artifacts.
- Two independent chambers for parallel experimental runs ensure high throughput. With the multi-sensor upgrade, the Oxygraph-2k is able to do multiple samples as well as detect multiple molecules, including Ca2+, NO, H2S.
- For diagnosis of OXPHOS in living and permeabilized cells, small amounts of tissue from transgenic animal models, and human needle biopsies requires high resolution respirometry to obtain accurate results even at high sample dilution. Less than 1 mg of fresh muscle tissue or <500,000 cells are required per chamber (10-fold less compared to conventional instruments).
- Multiple substrate/ inhibitor titration protocols extend the information gained on various OXPHOS complexes in a single incubation.
- O2k has an optimum chamber design and uses glass chambers and titanium stoppers avoiding oxygen-permeable materials such as Perspex or Teflon-coated stirrers, used in other respirometry instruments.
- Instrument also includes integrated Peltier-temperature control and an automatic titration-injection micropump.
- great sensitivity (the limit of detection of oxygen flux is ±1 pmol s-1ml-1 over the entire experimental oxygen range, and oxygen levels can be measured accurately and maintained at steady state by automatic feedback control down to 5 nM or 0.0005 kPa )
- temperature stability (± 0.001 °C in the range of 2 °C to 47 °C, with highly homogenous temperature in the chamber surrounded by the constant-temperature copper block with well stirred medium)
- robust calibration procedure, quality controls and instrumental performance/validation tests