|
Β |
(26 intermediate revisions by 9 users not shown) |
Line 1: |
Line 1: |
| | {{OROBOROS header page name}} |
| | |
| {{Publication | | {{Publication |
| |title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] An experiment with high-resolution respirometry: coupling control in cell respiration. Β | | Β |
| |info=[[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/a/a6/MiPNet10.04_CellRespiration.pdf |Bioblast pdf]] Β | | |title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] An experiment with high-resolution respirometry: coupling control in cell respiration. |
| |authors=OROBOROS | | |info=[[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/a/a6/MiPNet10.04_CellRespiration.pdf |Bioblast pdf]]Β Β»[http://www.bioblast.at/index.php/File:MiPNet10.04_CellRespiration.pdf Versions] |
| | |authors=Oroboros |
| |year=2016-08-08 | | |year=2016-08-08 |
| |journal=Mitochondr Physiol Network | | |journal=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»[http://www.bioblast.at/index.php/File:MiPNet10.04_CellRespiration.pdf Versions] | | |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 |
|
| |
|
| Methodological and conceptual features of highresolution respirometry are illustrated in an experiment with cultured, suspended cells in the OROBOROS O2k. The experiment demonstrates manual [[MiPNet09.12 O2k-Titrations |titrations]] of inhibitors, and automatic titrations of an uncoupler using the electronic [[O2k-Catalogue: TIP2k |Titration-Injection microPump TIP2k]]. Application of the DatLab 7 software is shown for instrumental control (O2k and TIP2k), and [[MiPNet19.18E O2 Flux Analysis |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).
| | {{MiPNet pdf page linking to MitoPedia}} |
| :>> Product: [http://www.oroboros.at/?oxygraph OROBOROS O2k], [[OROBOROS O2k-Catalogue | O2k-Catalogue]] | | :Β» Product: [[Oroboros O2k]], [[Oroboros O2k-Catalogue | O2k-Catalogue]] |
| |mipnetlab=AT Innsbruck OROBOROS | | Β |
| | |mipnetlab=AT Innsbruck Oroboros |
| }} | | }} |
| {{Labeling | | {{Labeling |
Line 17: |
Line 22: |
| |preparations=Intact cells | | |preparations=Intact cells |
| |topics=Coupling efficiency;uncoupling, Uncoupler | | |topics=Coupling efficiency;uncoupling, Uncoupler |
| |couplingstates=LEAK, ROUTINE, ETS | | |couplingstates=LEAK, ROUTINE, ET |
| |substratestates=ROX | | |pathways=ROX |
| |instruments=Oxygraph-2k, TIP2k, O2k-Protocol | | |instruments=Oxygraph-2k, TIP2k, O2k-Protocol |
| |additional=O2k-Demo, O2k-Core
| |
| }} | | }} |
| [[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]]
| |
|
| |
| ::Chemical Background Excel demo and template: [[Media:MiPNet10.04_CellRespiration O2k-ChemBackground.xlsx|MiPNet10.04_CellRespiration O2k-ChemBackground.xlsx]]
| |
|
| |
| ::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 ==
| |
|
| |
| * [[Gnaiger 2014 MitoPathways|References]]
| |
| * [[Coupling control protocol]]
| |
|
| |
|
| |
| [[Category:OroboPedia]]
| |
MiPNet10.04 CellRespiration
An experiment with high-resolution respirometry: coupling control in cell respiration.
|
Β» Β»Versions
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.
- Open the pdf document above.
- Β» 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, ET
Pathway: ROX
HRR: Oxygraph-2k, TIP2k, O2k-Protocol