Difference between revisions of "MiPNet10.04 CellRespiration"
Line 1: | Line 1: | ||
{{Publication | {{Publication | ||
|title=[[Image:O2k-Protocols.jpg|right|80px|link= | |title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] An experiment with high-resolution respirometry: Coupling control in cell respiration. | ||
|info=[[Media:MiPNet10.04 CellRespiration.pdf|'''Open Access-MiPNet10.04.pdf''']]; [http://www.bioblast.at/index.php/File:MiPNet10.04_CellRespiration.pdf Versions] | |info=[[Media:MiPNet10.04 CellRespiration.pdf|'''Open Access-MiPNet10.04.pdf''']]; [http://www.bioblast.at/index.php/File:MiPNet10.04_CellRespiration.pdf Versions] | ||
|authors=OROBOROS | |authors=OROBOROS | ||
|year= | |year=2015-01 | ||
|journal=Mitochondr Physiol Network | |journal=Mitochondr Physiol Network | ||
|abstract='''Gnaiger E ( | |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 Oxygraph-2k (O2k). The experiment demonstrates manual [ | Methodological and conceptual features of highresolution respirometry are illustrated in an experiment with cultured, suspended cells in the OROBOROS Oxygraph-2k (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 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 Oxygraph-2k Manual. The experiments were carried out by participants of an O2k-Workshop on high-resolution respirometry in April 2005 (IOC30; Schröcken, Austria). | ||
:>> Product: [http://www.oroboros.at/?oxygraph OROBOROS Oxygraph-2k], [[O2k-Catalogue_OROBOROS| O2k-Catalogue]] | :>> Product: [http://www.oroboros.at/?oxygraph OROBOROS Oxygraph-2k], [[O2k-Catalogue_OROBOROS| O2k-Catalogue]] | ||
|mipnetlab=AT_Innsbruck_OROBOROS}} | |mipnetlab=AT_Innsbruck_OROBOROS}} | ||
Line 20: | Line 20: | ||
|instruments=Oxygraph-2k, TIP2k, Protocol | |instruments=Oxygraph-2k, TIP2k, Protocol | ||
|additional=O2k-Demo, O2k-Core}} | |additional=O2k-Demo, O2k-Core}} | ||
[[Image:MiPNet10.04.jpg|center|width=80px|frame|Figure 1. Respiration of 32D cells (1.1106 ml-1). Traces for the two chambers of the Oxygraph-2k are superimposed. A: Oxygen flow [pmol | [[Image:MiPNet10.04.jpg|center|width=80px|frame|Figure 1. Respiration of 32D cells (1.1106 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.]] | ||
== Further information == | == Further information == | ||
:>> [http://www.oroboros.at/?protocols_cell_pcp Excel Templates and DatLab-Demo Files] | :>> [http://www.oroboros.at/?protocols_cell_pcp Excel Templates and DatLab-Demo Files] | ||
* [[Gnaiger | * [[Gnaiger 2014 MitoPathways|References]] | ||
* [[Coupling control protocol]] | |||
[[Category:OroboPedia]] | [[Category:OroboPedia]] |
Revision as of 12:29, 11 January 2015
An experiment with high-resolution respirometry: Coupling control in cell respiration. |
» Open Access-MiPNet10.04.pdf; Versions
OROBOROS (2015-01) 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 Oxygraph-2k (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 Oxygraph-2k Manual. The experiments were carried out by participants of an O2k-Workshop on high-resolution respirometry in April 2005 (IOC30; Schröcken, Austria).
- >> Product: OROBOROS Oxygraph-2k, 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, O2k-Core