Difference between revisions of "MiPNet14.06 Instrumental O2 background"

Latest revision as of 17:19, 19 October 2023

MiPNet14.06 Instrumental O2 background

O2k Quality Control 2: Instrumental oxygen background correction and accuracy of oxygen flux.

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Oroboros (2023-10-19) Mitochondr Physiol Network

Abstract: Timón-Gómez A, Grings M, Baglivo E, Schmitt S, Gnaiger E (2023) O2k Quality Control 2: Instrumental oxygen background correction and accuracy of oxygen flux. Mitochondr Physiol Network 14.6(09):1-16.

Open the pdf document above.

» Current O2k-series: NextGen-O2k Series XB and O2k Series J

» Current software versions DatLab 8.0: MitoPedia: DatLab

Further details: » MitoPedia: O2k-Open Support

» Product: O2k-FluoRespirometer, O2k-Catalogue

• O2k-Network Lab: AT_Innsbruck_Oroboros

Keywords

»Bioblast links: Chamber volume - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>

» Volume

Units: Liter [L]; 1 L = 1 dm³ = 10^-3 m³; 1 mL = 1 cm³; 1 µL = 1 mm³

» O2k-chamber

» The experimental chamber in the instrumental O2k-Chamber

» O2k-Chamber sV

» Calibration of O2k-chamber volume

On chamber volume

» O2k-chamber

» Chamber volume

» MiPNet09.01 O2k-ParadigmShift

» Cell count and normalization in HRR

» Open chamber

» Closed chamber

Components of the experimental O2k-chambers

» Stirrer-Bar\white PVDF\15x6 mm

» Stirrer-Bar sV\white PVDF\11.5x6.2 mm

» Stopper\black PEEK\conical Shaft\central Port

» Stopper sV\black PEEK\conical Shaft\central Port

» O-ring\Viton\12.5x1 mm

» O-ring sV\Viton\9.5x1 mm

» Volume-Calibration Ring

» Volume-Calibration Ring sV

» Stopper-Spacer

» Cover-Slip\black

» O2k-Chamber Holder

» O2k-Chamber Holder sV

» OroboPOS-Holder

» OroboPOS-Holder sV

Maintencance

» O2k-cleaning and ISS

» Quality control: O2k-chamber test

Acknowledgements

Labels: MiParea: Respiration, Instruments;methods

HRR: Oxygraph-2k, O2k-Manual

DatLab, O2k-SOP, DL7, System, MitoFit2022QC

@@ Line 1: / Line 1: @@
+{{Template:OROBOROS support page name}}
 {{Publication
-|title=Fasching M, Gnaiger E. Instrumental background correction and accuracy of oxygen flux. Mitochondr Physiol Network 14.6.
+|title=[[Image:O2k-Manual.jpg|right|70px|link=O2k-Manual|O2k-Manual]] O2k Quality Control 2: Instrumental oxygen background correction and accuracy of oxygen flux.
-|info=[http://www.oroboros.at/index.php?backgroundcorrection www.oroboros.at/Standard Operating Procedures]
+|info=[[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/6/65/MiPNet14.06_InstrumentalO2Background.pdf |Bioblast pdf]] » [http://www.bioblast.at/index.php/File:MiPNet14.06_InstrumentalO2Background.pdf Versions]
-|authors=Fasching M, Gnaiger E
+|authors=Oroboros
-|year=*
+|year=2023-10-19
 |journal=Mitochondr Physiol Network
-|abstract=For calibration of the polarographic oxygen sensor (POS) and for measurement of instrumental background oxygen consumption, only incubation medium but no biological sample is added into the Oxygraph-2k chamber, at experimental conditions. In a closed chamber under these conditions, ideally oxygen concentration remains constant. In practice, however, instrumental background effects are caused by back-diffusion into the oxygraph chamber at low oxygen pressure, oxygen diffusion out of the oxygraph chamber at elevated oxygen levels, and oxygen consumption by the polarographic oxygen sensor (POS). Instrumental background interferes with accurate measurement of respiratory oxygen flux, if background effects remain undefined. Determination of instrumental background constitutes an important standard operating procedure (SOP) in high-resolution respirometry (HRR) Instrumental background oxygen flux is (i) minimized in the OROBOROS Oxygraph-2k by instrumental design and selection of appropriate materials. In addition, (ii) instrumenal background is routinely tested, and (iii) background correction of oxygen flux is applied automatically by DatLab.
+|abstract=Timón-Gómez A, Grings M, Baglivo E, Schmitt S, Gnaiger E (2023) O2k Quality Control 2: Instrumental oxygen background correction and accuracy of oxygen flux. Mitochondr Physiol Network 14.6(09):1-16.
-|keywords=Instrumental background
+{{MiPNet pdf page linking to MitoPedia}}
-|mipnetlab=AT_Innsbruck_OROBOROS
+:» Product: [[O2k-FluoRespirometer]], [[Oroboros O2k-Catalogue |O2k-Catalogue]]
-|discipline=Mitochondrial Physiology
+|mipnetlab=AT_Innsbruck_Oroboros
-|articletype=Protocol; Manual, MiPNet-online Publication
 }}
-{{Labeling
-|instruments=Oxygraph-2k, Protocol, MiPNet-Publication
-|additional=O2k-SOP
-|discipline=Mitochondrial Physiology
-|articletype=Protocol; Manual, MiPNet-online Publication
-}}
-== Instrumental background test for permeabilized muscle fibres ==
-=== Extended instrumental background test ===
+== Keywords ==
+{{Template:Keywords: Chamber volume}}
-# While biopsy sampling and fibre preparation proceed: Air calibration in [[MiR06Cr]], then close the chamber to evaluate instrumental background at air saturation (c. 10 min): This is a quality control of the medium, important under field conditions, where medium preservation (sterility) may be less controlled than in the lab.
-# Elevate oxygen concentration to 450 µM with oxygen gas ([[Syringe\60 ml\Gas-Injection]]), close and after two to three min perform a [[stirrer test]] (the new DatLab has an automatic stirrer test function, with defined duriation of stopping the stirrer and automatic restart of stirring, 40 s stirrer stop may be optimum). This is important, since the [[OroboPOS]] may have a different [[response time]] at elevated oxygen concentration. If the response time increases dramatically, then the sensor may even show a non-linear response to oxygen concentration at high oxygen levels.
-# Instrumental background: After 20 min, open the chamber and allow O<sub>2</sub> to drop to c. 350 µM, close for 20 min, open and drop O<sub>2</sub> to c. 250 µM (this should be the lowest experimental O<sub>2</sub> concentration).
-# Increase O<sub>2</sub> with H<sub>2</sub>O<sub>2</sub> injection (c. 2 µl) to 400 µM, measure for 15-20 min instrumental background, simulating a re-oxygenation during the experiment.
-# Increase O<sub>2</sub> with H<sub>2</sub>O<sub>2</sub> injection (c. 1 µl) to 450 µM, until the fibres are added, for equilibrating the instrument at high O<sub>2</sub>.
-# Addition of permeabilized fibres into the O2k-Chamber: >> [[Permeabilized muscle fibres]]
-==Info==
+'''Acknowledgements'''
+[[File:Template NextGen-O2k.jpg|left|400px|link=NextGen-O2k]]
+<br/>
+<br/>
+<br/>
+<br/>
-MiPNet10.07 is integrated in MiPNet14.06.
+{{Labeling
+|area=Respiration, Instruments;methods
+|instruments=Oxygraph-2k, O2k-Manual
+|additional=DatLab, O2k-SOP, DL7, System, MitoFit2022QC
+}}