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| == Leaky Chamber ==
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| === Leaky chamber test ===
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| :::* When the leakage is so little that cannot be noticed in a short amount of time, we recommend to follow the ''Leaky chamber test'' below:
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| ::::# Decrease the oxygen concentration down to ~ 50 ฮผM (using [[Dithionite]] or N<sub>2</sub> injection)
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| ::::# Let the O2k record overnight with the [[closed chamber|chamber closed]] and illumination switched off
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| ::::# The oxygen concentration should decrease overtime due to the O<sub>2</sub> consumed by [[OroboPOS]] as it is shown in the figure. In this case, the chamber is assembledย properly.
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| ::::# If the oxygen concentration increases over time, a reassemble of the chamber is needed.
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| [[File:Leaking_chamber_2020-02-03 P2-03.png|1500px]]
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| ::::*'''Text in discussion for the page:'''
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| ::::* To exclude the existence of an unnoticed leak from the chamber, we recommend performing the following ''Leaky chamber test'' - especially after chamber reassembly:
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| ::::# Decrease the oxygen concentration down to ~ 50 ฮผM (using [[Dithionite]] or N<sub>2</sub> injection).
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| ::::# Record the O2k measurement overnight with the [[closed chamber|chamber closed]] and illumination switched off.
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| ::::# The oxygen concentration should decrease overtime due to the O<sub>2</sub> consumed by the [[OroboPOS]] as shown in the figure below.
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| ::::# If the oxygen concentration increases over time, a reassemblye of the chamber is needed.
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| ::::[[User:CecattoC|CecattoC]] ([[User talk:CecattoC|talk]]) 15:52, 19 March 2020 (CET). So, it should be done every reassemble otherwise nobody will noticed that? How frequent you received cases with this or noticed leaky chamber in our lab? If the user can see the decrease of oxygen consumed by the sensor in an half and hour or an hour closed chamber run, they don't need to perform the leaky chamber test, right?
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| === A story about gas bubbles appearing in the closed chamber ===
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| [[User:Gnaiger Erich|Gnaiger Erich]] ([[User talk:Gnaiger Erich|talk]]) 16:27, 19 March 2020 (CET)
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| ::::* You are an O2k user who assembles the O2k-Chamber carefully following the SOP (manual & video). Usually this works very well - but not always ..
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| ::::# You are stressed because you have a persistent problem with a leak after chamber assembly, loosing medium from the chamber (observed by medium dropping out from the OroboPOS holder and connector; or a gas bubble appearing slowly in the closed chamber).
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| ::::# โLeaky chamberโ, therefore, has to be defined: Here it is not just the oxygen leak (as observed in an instrumental O2-background test), it is actual medium leak โ two entirely different processes, although medium leak causes simultaneously an O2 leak, but not the other way around.
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| ::::# You re-assemble again and again, until the chamber appears to be tight. You have no clue, what you did differently. And you are not 100 % sure, if the chamber is now really tight. (This is meant by โWhen the leakage is so little that cannot be noticedโ).
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| ::::# You could test this easily with an instrumental O<sub>2</sub>-background test - SOP! You can do this even in a short amount of time, if you have other tasks to complete meanwhile. But unfortunately it is already evening, and the lab will be closed. (This is meant by โin a short amount of timeโ). Well, then you may quickly reduce the oxygen level .. the overnight โleaky chamber testโ (we will find a better title).
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| ::::# You may use dithionite quickly. But probably you performed your tests of loosing medium after chamber assembly with H<sub>2</sub>O, to avoid any sticky medium clogging your sensor connector when medium leaks from the chamber. Do you now titrate dithionite solution into the closed chamber containing H<sub>2</sub>O? YOU WOULD BE IN TROUBLE! This is well explained in our SOPs, but it is a good idea to clarify, that you should use MiR05 (or a highly concentrated phosphate buffer). (Marco: did you actually use MiR05 โ I guess absolutely YES).
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| ::::# You come back to the lab in the morning โ BINGO. The chamber is tight. Does this replace the instrumental O<sub>2</sub>-background test?
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