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| '''''Do I have to stop a run, when oxygen is approaching to zero and becomes limiting?'''''
| | #REDIRECT [[MiPNet14.13_Medium-MiR06#Re-oxygenation_with_H2O2_titrations]] |
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| An experiment is not finished, because of running out of oxygen. There are different possibilities to re-oxygenate.
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| *'''Re-oxygenation by adding H2O2 to a catalase containing medium ([[MiPNet14.13 Medium-MiR06]])'''
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| :Add catalase at a final concentration of 280 IU/ml to the medium at the beginning of an experiment. When oxygen starts to become limited, inject 1-3 µl of an approximately 200 mM H2O2 stock solution. The H2O2 will immediately be degraded to O2; the catalase concentration is high enough to avoid any oxidative stress. In this approach the chamber is not opened and closed, so the disturbance of the system is less and the stabilization phase of the sensor is short compared to a re-oxygenatio nby opening the chamber.
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| *'''Re-oxygenation by opening the chamber'''
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| :Lift the stopper and leave a gas volume above the liquid phase (use the stopper-spacer tool to set the stopper in the right position). Leave the chamber open till oxygen is again up to approximately air calibration level. Close the chamber by inserting the stopper completely and wait till the sensor is stable again (may take 5 to 10 minutes).
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| If possible, it is preferable to re-oxygenate in a phase of low respiratory activity, only little amounts of oxygen are consumed during the stabilization phase of the sensor after closing the chamber.
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| If you replace the air phase above the liquid phase by pure oxygen you can increase oxygen levels above air saturation, as it is recommended for measuring mitochondrial respiratory function in muscle biopsies.
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| ==See also==
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| * [[MiR06]]
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