Difference between revisions of "MiPNet10.04 CellRespiration"

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{{Publication
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|title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] An experiment with high-resolution respirometry: coupling control in cell respiration.
 
|title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] An experiment with high-resolution respirometry: coupling control in cell respiration.
 
|info=[[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/a/a6/MiPNet10.04_CellRespiration.pdf |Bioblast pdf]]  »[http://www.bioblast.at/index.php/File:MiPNet10.04_CellRespiration.pdf Versions]
 
|info=[[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/a/a6/MiPNet10.04_CellRespiration.pdf |Bioblast pdf]]  »[http://www.bioblast.at/index.php/File:MiPNet10.04_CellRespiration.pdf Versions]
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|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.'''
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|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.
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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 - DatLab 6 |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; Schröcken, Austria).
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{{MiPNet pdf page linking to MitoPedia}}
 
:» Product: [[Oroboros O2k]], [[Oroboros O2k-Catalogue | O2k-Catalogue]]
 
:» Product: [[Oroboros O2k]], [[Oroboros O2k-Catalogue | O2k-Catalogue]]
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|mipnetlab=AT Innsbruck Oroboros
 
|mipnetlab=AT Innsbruck Oroboros
 
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|pathways=ROX
 
|pathways=ROX
 
|instruments=Oxygraph-2k, TIP2k, O2k-Protocol
 
|instruments=Oxygraph-2k, TIP2k, O2k-Protocol
|additional=O2k-Demo, O2k-Core, SUIT-003 O2 ce D009, SUIT-003, SUIT-006 General O2 pce D053,
 
 
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[[Image:MiPNet10.04.jpg|left|500px|]] Respiration of 32D cells (1·10<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>·s<sup>-1</sup>·10<sup>6</sup>];  B: Oxygen concentration [μM] (graph layout “05b Specific flux  overlay”). After inhibition of ATP synthase with oligomycin, FCCP was  titrated at 0.5 μM 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.
 
 
 
== Cell coupling control protocol ==
 
****: ceCCP: [[1ce;2ceOmy;3ceU-]]
 
 
== DatLab-Analysis 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-Analysis template: [[Media:SUIT_MiPNet10.04_CellRespiration.xlsx|SUIT_MiPNet10.04_CellRespiration.xlsx]]
 
 
== Further information ==
 
 
::::» [[Gnaiger 2008 POS]]
 
::::» [[Gnaiger 2014 MitoPathways|References]]
 
::::» [[Coupling control protocol]]
 
::::» [[DatLab]]
 
 
[[Category:OroboPedia]]
 

Latest revision as of 11:51, 4 December 2019

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MiPNet10.04 CellRespiration


Publications in the MiPMap
O2k-Protocols
An experiment with high-resolution respirometry: coupling control in cell respiration.

» Bioblast pdf  »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.



O2k-technical support and open innovation
Open the pdf document above.
» Current O2k-series: O2k-FluoRespirometer
» Current software version: DatLab 7
» Product: Oroboros O2k, O2k-Catalogue


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