MiPNet10.04 CellRespiration: Difference between revisions
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::::Β» [[Gnaiger 2014 MitoPathways|References]] | ::::Β» [[Gnaiger 2014 MitoPathways|References]] | ||
::::Β» [[Coupling control protocol]] | ::::Β» [[Coupling control protocol]] | ||
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[[Category:OroboPedia]] | [[Category:OroboPedia]] |
Revision as of 09:50, 9 May 2017
An experiment with high-resolution respirometry: coupling control in cell respiration. |
OROBOROS (2016-08-08) Mitochondr Physiol Network
Abstract: Doerrier C, Gnaiger E (2005-2016) An experiment with high-resolution respirometry: Coupling control in cell respiration. Mitochondr Physiol Network 10.04(09):1-12. Β»Versions
Methodological and conceptual features of highresolution respirometry are illustrated in an experiment with cultured, suspended cells in the OROBOROS O2k. The experiment demonstrates manual titrations of inhibitors, and automatic titrations of an uncoupler using the electronic Titration-Injection microPump TIP2k. Application of the DatLab 7 software is shown for instrumental control (O2k and TIP2k), and real-time data analysis. The following guideline describes the experiment in the form of a laboratory protocol, complementary to the relevant sections of the O2k Manual. The experiments were carried out by participants of an O2k-Workshop on high-resolution respirometry in April 2005 (IOC30; SchrΓΆcken, Austria).
- Β» Product: Oxygraph-2k, O2k-Catalogue
β’ O2k-Network Lab: AT Innsbruck OROBOROS
Labels: MiParea: Respiration, Instruments;methods
Organism: Mouse
Tissue;cell: Blood cells
Preparation: Intact cells
Regulation: Coupling efficiency;uncoupling, Uncoupler Coupling state: LEAK, ROUTINE, ETS"ETS" is not in the list (LEAK, ROUTINE, OXPHOS, ET) of allowed values for the "Coupling states" property. Pathway: ROX HRR: Oxygraph-2k, TIP2k, O2k-Protocol
O2k-Demo, O2k-Core, CCP02
Respiration of 32D cells (1Β·106 ml-1). Traces for the two chambers of the O2k are superimposed. A: Oxygen flow [pmol O2Β·s-1Β·106]; B: Oxygen concentration [ΞΌM] (graph layout β05b Specific flux overlayβ). After inhibition of ATP synthase with oligomycin, FCCP was titrated at 0.5 ΞΌM steps with the TIP2k. Each titration step is automatically marked by an event (vertical lines). After the aerobic-anoxic transition, the chambers were opened for re-oxygenation, causing a 10-min disturbance of the traces of respiratory flow. The TIP2k needles were removed before opening, and re-inserted after closing the chamber.
Coupling control protocol
- CCP: CCP02
DatLab-Analysis templates and DatLab Demo Files
DatLab 7
- Calibration-File: MiPNet10.04_CellRespiration_Calib.DLD
- Demo-File: MiPNet10.04_CellRespiration.DLD
- Chemical Background Excel demo and template: MiPNet10.04_CellRespiration O2k-ChemBackground.xlsx
- DatLab-Analysis template: SUIT_MiPNet10.04_CellRespiration.xlsx
Further information
- Β» Gnaiger 2008 POS
- Β» References
- Β» Coupling control protocol
- Β» DatLab