Difference between revisions of "Talk:Oxygen calibration - DatLab"

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* [[Air_calibration|Air calibration]]
* [[Air_calibration|Air calibration]]
* [[Oxygen_sensor_test|Oxygen sensor test]]
* [[Run_DL-Protocol/Set_O2_limit|Run DL-Protocol]]
* [[Open_chamber|Open chamber]]
* [[Open_chamber|Open chamber]]
* [[Closed_chamber|Closed chamber]]
* [[Closed_chamber|Closed chamber]]
Line 71: Line 73:
* Quality control: [[Oxygen_sensor_test]]
* Quality control: [[Oxygen_sensor_test]]
* removing oxygen from the chamber with [[O2-Zero_Powder]]
* removing oxygen from the chamber with [[O2-Zero_Powder]]
* [[Oxygen_solubility_factor|Oxygen solubility factor]]
* [[Oxygen_flux_-_instrumental_background|Oxygen flux - instrumental background]]

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Talk:Oxygen calibration - DatLab

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O2k-Open Support

MitoPedia O2k and high-resolution respirometry: O2k-Open Support 


O2 calibration is the calibration in DatLab of the oxygen sensor. It is a prerequisite for obtaining accurate measurements of respiration. Accurate calibration of the oxygen sensor depends on (1) equilibration of the incubation medium with air oxygen partial pressure at the temperature defined by the experimenter; (2) zero oxygen calibration; (3) high stability of the POS signal tested for sufficiently long periods of time; (4) linearity of signal output with oxygen pressure in the range between oxygen saturation and zero oxygen pressure; and (5) accurate oxygen solubility for aqueous solutions for the conversion of partial oxygen pressure into oxygen concentration. The standard oxygen calibration procedure is described below for high-resolution respirometry with the calibration routine using instrumental calibration DL-Protocols in DatLab.

Reference: MiPNet06.03 POS-calibration-SOP, MiPNet12.20_O2k-calibration_tutorial

Oxygen calibration - when and how?

Air calibration / Quality control - R1

  • Daily air calibration goes hand in hand with a stirrer test and inspection of the recorded data in DatLab, also called oxygen sensor test. The procedure is decisive for maintaining a high level of O2k quality control as described in QC 1: Oxygen sensor test.
  • Use the air calibration protocol as a guide to generate R1. Main steps:
  • Record the oxygen signal while the chamber is filled with medium and the stopper is in the 'open chamber' position.
  • After equilibration set a mark and call it R1 (named automatically when the DatLab protocol is used)
O2 calibration indicator fields in DatLab 7
O2 calibration indicator fields in DatLab 8
Indicator fields
As a reminder, the calibration indicator fields in the bottom right and bottom left corner of the experiment window are colored yellow after starting an experiment. After opening the calibration window by pressing F5 (or via the top menu bar) and confirming the calibration, the field changes to green (in DatLab 8 as well as in DatLab 7.4).

Zero oxygen calibration - R0

  • Less frequent than air calibration, zero oxygen calibration should be done from time to time over weeks. In addition, zero oxygen calibration is recommended when the experimental workflow requires recording of low oxygen concentrations. It is mandatory after application of a new membrane and O2-sensor service to confirm sensor serviceability.
  • Use the 'air and zero' calibration protocol as a guide to generate R0. Main steps:
  • In the closed chamber remove the oxygen from the medium (chemically by adding sodium dithionite or biologically by adding oxygen consuming sample)
  • Set a mark and call it R0 (named automatically during the DatLab protocol)
Instrumental O2 background test
As described in more detail in MiPNet14.06_Instrumental_O2_background, the instrumental O2 background test is necessary to assess the oxygen fluxes into or out of the respirometer chamber that are not directly related to respiratory activity of the sample. Is is recommended to perform it about once a month. Air calibration and zero oxygen calibration are part of the procedure.

DatLab 8

O2 calibration window manual.png
O2 calibration window calib and active file.png
  • How to get R1 and R0
  • calibration values are stored together with instrument identifiers and sensor numbers
  • bottom left and bottom right: yellow until calibration has been confirmed

DatLab 7.4

DLP O2-calibration air and zero.png
Calibration window steps.png

Related Topics