Difference between revisions of "Talk:O2k-chamber"
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== Different fluxes in left and right chmaber == | |||
'''Question:''' In our experiments we get consistently higher fluxes in chamber A as compared to chamber B. | |||
'''Answer:''' To exclude instrumental causes please follow the procedure described in the section | |||
'''Calibration and quality control of the OroboPOS (O2k-SOP)''' in [[MiPNet06.03 POS-Calibration-SOP]]. | |||
Possible instrumental and protocol related causes for fluxes differing between chambers. | |||
* wrong O2 calibration in one chamber | |||
* wrong chamber volume calibration: The total amount of oxygen in the chamber with the smaller volume will be smaller. Therefore the O2 concentration will drop faster resulting in a higher flux. | |||
* wrong background parameters: wrong background parameters will cause a O2 concentration dependent offset to the flux, the value of the offset will be independent of the absolute value of the flux. E.g. a constant offset of 10 pmol /ml s at ca 100 µM O2 concentration and 20 pmol/ml s at 300 µM O2 concentration could be caused by wrong background parameters, but a difference of always 20% at very different absolute values of the flux can not be caused by wrong background parameters. In any case it is advisable to check the background parameters ( redo an instrumental O2 background experiment). | |||
* hydrophobic inhibitors | |||
* biological contamination |
Revision as of 11:00, 6 November 2014
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O2k-Chamber has been accessed more than
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Different fluxes in left and right chmaber
Question: In our experiments we get consistently higher fluxes in chamber A as compared to chamber B.
Answer: To exclude instrumental causes please follow the procedure described in the section Calibration and quality control of the OroboPOS (O2k-SOP) in MiPNet06.03 POS-Calibration-SOP.
Possible instrumental and protocol related causes for fluxes differing between chambers.
- wrong O2 calibration in one chamber
- wrong chamber volume calibration: The total amount of oxygen in the chamber with the smaller volume will be smaller. Therefore the O2 concentration will drop faster resulting in a higher flux.
- wrong background parameters: wrong background parameters will cause a O2 concentration dependent offset to the flux, the value of the offset will be independent of the absolute value of the flux. E.g. a constant offset of 10 pmol /ml s at ca 100 µM O2 concentration and 20 pmol/ml s at 300 µM O2 concentration could be caused by wrong background parameters, but a difference of always 20% at very different absolute values of the flux can not be caused by wrong background parameters. In any case it is advisable to check the background parameters ( redo an instrumental O2 background experiment).
- hydrophobic inhibitors
- biological contamination