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Difference between revisions of "MiPNet15.05 NO-manual"

From Bioblast
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|area=Instruments;methods
|area=Instruments;methods
|instruments=Oxygraph-2k, NO, O2k-Manual
|instruments=Oxygraph-2k, NO, O2k-Manual
|additional=O2k-MultiSensor, O2k-SOP, DL6, DL7, DL6a7
|additional=O2k-MultiSensor, DL6, DL7, DL6a7
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Revision as of 12:22, 6 September 2018

                



MiPNet15.05 NO-manual

Publications in the MiPMap
O2k-Manual
O2k-MultiSensor system with amperometric sensors: NO, H2S, H2O2.

Β» Bioblast pdf  Β»Versions

Oroboros (2014-03-06) Mitochondr Physiol Network

Abstract: Fasching M, Gnaiger E (2014) O2k-MultiSensor system with amperometric sensors: NO, H2S, H2O2. Mitochondr Physiol Network 15.05(04):1-8. - Versions


O2k-Manual: Two additional amperometric amplifiers are integrated into the OROBOROS O2k-Core. Two NO sensors can be directly connected, and both chambers can be used simultaneously for oxygen and NO, and two additional potentiometric electrodes can be monitored simultaneously. NO, H2O2 and similar sensors are amperometric sensors, as is the Oroboros polarographic oxygen sensor (OroboPOS). The measured current is converted to a voltage, amplified and recorded by DatLab.

Β» Product: O2k-NO Amp-Module, O2k-Catalogue


β€’ O2k-Network Lab: AT_Innsbruck_Oroboros


Labels: MiParea: Instruments;methods 





HRR: Oxygraph-2k, NO, O2k-Manual 

O2k-MultiSensor, DL6, DL7, DL6a7 



Template NextGen-O2k.jpg


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




O2k signal and output

  1. O2k signal: The O2k-NO Amp-Module is operated through the amperometric (Amp)-Channel of the O2k, with electric current (ampere [A]) as the primary signal.
  2. O2k output: type A and B


Electrochemical NO and H2O2 sensors

NO and H2O2 sensors use an amperometric measurement principle (Amp-Module), comparable to the polarographic oxygen sensor (OroboPOS), which yields an amperometric signal at a given polarization voltage. We supply custom-made stoppers that can accommodate such sensors. All amperometric sensors (NO, H2S, H2O2) that have the same connection can be used with the O2k.

Requirements for NO or other amperometric sensors

Figure 1. MultiSensor Features of DatLab 4: NO-Signal - Graph layout


1.NO sensor (not yet available from Oroboros Instruments; see Aguirre et al 2010).
2.PVDF stopper to accommodate the NO sensor (#30221-24 for 2 mm shaft NO sensor).


Oxygraph-2k Series A-C
A.3. NO amplification box (#30430-24).


Oxygraph-2k Series D (and higher)
D.3. MultiSensor electronic upgrading (#30100-34).


Questions.jpg

NO-sensor calibration

  • I have a question regarding NO-sensor calibration. What method would you recommend? I just tried with a solution (outside of the Oroboros O2k) recommended from the factory (0.05 M H2SO4, 0.02 M KI with addition of nitrite in different concentrations). This method worked fine, however, is this possible to do in the O2k? Is that extremely acidic solution harmful to the POS membranes? Best regards ~ Schiffer TA
  1. Answer: That is the method we always used. It is fine for the O2k and no problem for the PTFE O2 membranes. But I have to admit that the laboratory table got some "scars" :-) from the KI, and the blue POM chamber holders may change colour. ~ support@oroboros.at, 10 March 2011


Questions.jpg

Pre-polarisation times

  • For some sensors WPI states the suggested pre-polarisation time not in the manual of their sensor but only in the manual of the WPI electronics.
  • User feedback for the WPI HPO-2 sensor: The suggested polarization voltage for the WPI H2O2 electrodes is 400 mV, that means -400 mV have to be set in the O2k (see MiPNet15.05 NO-Manual). I have also contacted with WPI. They said that "The ISO-HPO-2 polarized at 450 mV for 2 hours in 0.1 M PBS solution." But I find 2 hours is not enough. I send this information to you, maybe it is helpful for other customers.
  1. Comment by Oroboros: Based on our experience with the WPI NO electrode and our users experience with the WPI HPO electrode, the required prepolarisation time is longer than the (minimum) cited by WPI. Especially for the first run we suggest to allow for a very long pre-polarisation time (minimum over-night, days for NO) and to observe the slope of the signal. Only when the slope of the signal (not the signal itself !) is constant for a long time (hours), pre-polarisation is finished. This will give an indication for the proper pre-polarisation time in further experiments. It is important to continue the pre-polarisation for a few hours after the slope appears to be constant. In the later phase of pre-polarisation the slope may change very slowly but may still approach closer to zero. This can only be detected by recording the slope for a sufficiently long period of time. ~ support@oroboros.at, 23 January 2014