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Difference between revisions of "MiPNet10.04 CellRespiration"

From Bioblast
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|substratestates=ROX
|substratestates=ROX
|instruments=Oxygraph-2k, TIP2k, Protocol
|instruments=Oxygraph-2k, TIP2k, Protocol
|additional=O2k-Demo, O2k-Core
|additional=O2k-Demo
}}
}}
[[Image:MiPNet10.04.jpg|center|width=80px|frame|Figure 1. Respiration of 32D cells (1.1106 ml-1). Traces for the two chambers of the Oxygraph-2k are superimposed. A: Oxygen flow [pmol O<sub>2</sub>·s<sup>-1</sup>·10<sup>6</sup>]; B: Oxygen concentration [μM] (graph layout “7 Gr1-Flux Gr2-O2 Conc”). 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.]]
[[Image:MiPNet10.04.jpg|center|width=80px|frame|Figure 1. Respiration of 32D cells (1.1106 ml-1). Traces for the two chambers of the Oxygraph-2k are superimposed. A: Oxygen flow [pmol O<sub>2</sub>·s<sup>-1</sup>·10<sup>6</sup>]; B: Oxygen concentration [μM] (graph layout “7 Gr1-Flux Gr2-O2 Conc”). 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.]]

Revision as of 15:35, 19 May 2015

Publications in the MiPMap
O2k-Protocols
An experiment with high-resolution respirometry: Coupling control in cell respiration. »Bioblast pdf«

» Versions

OROBOROS (2015-01-11) Mitochondr Physiol Network

Abstract: Gnaiger E (2015) An experiment with high-resolution respirometry: Coupling control in cell respiration. Mitochondr Physiol Network 10.04(08):1-12.

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 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; Schröcken, Austria).

>> 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, ETS"ETS" is not in the list (LEAK, ROUTINE, OXPHOS, ET) of allowed values for the "Coupling states" property. 

HRR: Oxygraph-2k, TIP2k, Protocol"Protocol" is not in the list (Oxygraph-2k, TIP2k, O2k-Fluorometer, pH, NO, TPP, Ca, O2k-Spectrophotometer, O2k-Manual, O2k-Protocol, ...) of allowed values for the "Instrument and method" property. 

O2k-Demo 

Figure 1. Respiration of 32D cells (1.1106 ml-1). Traces for the two chambers of the Oxygraph-2k are superimposed. A: Oxygen flow [pmol O2·s-1·106]; B: Oxygen concentration [μM] (graph layout “7 Gr1-Flux Gr2-O2 Conc”). 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.

Further information

>> Excel Templates and DatLab-Demo Files