MiPNet10.04 CellRespiration: Difference between revisions
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[[Image:MiPNet10.04.jpg|center|width=80px|frame|Figure 1. Respiration of 32D cells (1Β·10<sup>6</sup> ml<sup>-1</sup>). Traces for the two chambers of the O2k are superimposed. A: Oxygen flow [pmol O<sub>2</sub>Β·s<sup>-1</sup>Β·10<sup>6</sup>]; 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.]] | [[Image:MiPNet10.04.jpg|center|width=80px|frame|Figure 1. Respiration of 32D cells (1Β·10<sup>6</sup> ml<sup>-1</sup>). Traces for the two chambers of the O2k are superimposed. A: Oxygen flow [pmol O<sub>2</sub>Β·s<sup>-1</sup>Β·10<sup>6</sup>]; 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.]] | ||
== DatLab-Excel templates and DatLab Demo Files == | |||
=== DatLab 7 === | |||
::Calibration-File: [[Media:MiPNet10.04_CellRespiration_Calib.DLD|MiPNet10.04_CellRespiration_Calib.DLD]] | |||
::Demo-File: [[Media:MiPNet10.04_CellRespiration.DLD|MiPNet10.04_CellRespiration.DLD]] | |||
::Background Excel demo and template: [[Media:MiPNet10.04_CellRespiration O2-Background.xls|MiPNet10.04_CellRespiration O2-Background.xls]] | |||
::Chemical Background Excel demo and template: [[Media:MiPNet10.04_CellRespiration O2k-ChemBackground.xls|MiPNet10.04_CellRespiration O2k-ChemBackground.xls]] | |||
::DatLab-Excel template: [[Media:SUIT_MiPNet10.04_CellRespiration.xlsx|SUIT_MiPNet10.04_CellRespiration.xlsx]] | |||
=== DatLab 6 === | |||
::Calibration-File: [[Media:MIPNET08.09 2003-03-29 P1-01 CALIB.DLD|MIPNET08.09_2003-03-29_P1-01_CALIB.DLD]] | |||
::Demo-File: [[Media:MIPNET08.09 2003-03-29 P1-02 CELLS.DLD|MIPNET08.09_2003-03-29_P1-02_CELLS.DLD]] | |||
::Background Excel demo and template: [[Media:MiPNet08.09 O2k-Background Cells DL6.xlsx|MiPNet08.09_O2k-Background_Cells.xlsx]] | |||
::DatLab-Excel template: [[Media:MiPNet08.09 O2k-Analysis Cells DL6.xlsx|MiPNet08.09_O2k-Analysis_Cells.xlsx]] | |||
=== DatLab 5 === | |||
::Calibration-File: [[Media:MIPNET08.09 2003-03-29 P1-01 CALIB.DLD|MIPNET08.09_2003-03-29_P1-01_CALIB.DLD]] | |||
::Demo-File: [[Media:MIPNET08.09 2003-03-29 P1-02 CELLS.DLD|MIPNET08.09_2003-03-29_P1-02_CELLS.DLD]] | |||
::Background Excel demo and template: [[Media:O2k-Background Cells 0809 DL5.xlsx|O2k-Background_Cells_0809.xlsx]] | |||
::DatLab-Excel template: [[Media:O2k-Analysis Cells 0809 DL5.xlsx|O2k-Analysis_Cells_0809.xlsx]] | |||
== Further information == | == Further information == | ||
* [[Gnaiger 2014 MitoPathways|References]] | * [[Gnaiger 2014 MitoPathways|References]] |
Revision as of 16:42, 9 August 2016
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).
- >> Product: OROBOROS O2k, 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.
HRR: Oxygraph-2k, TIP2k, O2k-Protocol
O2k-Demo, O2k-Core
DatLab-Excel templates and DatLab Demo Files
DatLab 7
- Calibration-File: MiPNet10.04_CellRespiration_Calib.DLD
- Demo-File: MiPNet10.04_CellRespiration.DLD
- Background Excel demo and template: MiPNet10.04_CellRespiration O2-Background.xls
- Chemical Background Excel demo and template: MiPNet10.04_CellRespiration O2k-ChemBackground.xls
- DatLab-Excel template: SUIT_MiPNet10.04_CellRespiration.xlsx
DatLab 6
- Calibration-File: MIPNET08.09_2003-03-29_P1-01_CALIB.DLD
- Demo-File: MIPNET08.09_2003-03-29_P1-02_CELLS.DLD
- Background Excel demo and template: MiPNet08.09_O2k-Background_Cells.xlsx
- DatLab-Excel template: MiPNet08.09_O2k-Analysis_Cells.xlsx
DatLab 5
- Calibration-File: MIPNET08.09_2003-03-29_P1-01_CALIB.DLD
- Demo-File: MIPNET08.09_2003-03-29_P1-02_CELLS.DLD
- Background Excel demo and template: O2k-Background_Cells_0809.xlsx
- DatLab-Excel template: O2k-Analysis_Cells_0809.xlsx