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| ==Problem==
| | #REDIRECT [[MiPNet19.03 O2k-cleaning and ISS]] |
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| Lasting effect of inhibitors (i.e. rotenone, antimycin A), as observed by persistent low fluxes, when working with permeabilized fibers.
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| ==Cleaning the Chamber==
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| '''Clean the chamber after an experiment involving lipid-soluble inhibitors (such as oligomycin, rotenone, or antimycin A):'''
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| *The chamber must be cleaned rigorously with ethanol (100%), since such inhibitor(s) are difficult to be washed out from the chamber and may inhibit mitochondrial respiration in subsequent experiments.
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| *Siphon off the cell/mitochondrial suspension at the end of the experiment and rinse the chamber with distilled water three times, by filling the chamber up to the rim.
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| *Also rinse the surface and capillary of the stopper with distilled water properly.
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| *Fill the water-cleaned chamber with 70 % ethanol and replace the stopper making sure that the ethanol fills up the receptacle, and leave for 5 min.
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| *Remove the stopper and siphon off the ethanol to empty the chamber. Repeat these cleaning steps with 70 % ethanol three times.
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| *Then fill the chamber with absolute ethanol (99.6 %) and insert the stopper making sure that the ethanol fills up the receptacle. Place the perspex cover on top of the stopper and leave for 15-20 min.
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| *At the end of this rigorous cleaning procedure remove the stopper and place it inverted (with the receptacle at the bottom) on a clean paper towel.
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| *Siphon off the ethanol from the chamber and rinse it with distilled water three times.
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| *Rinse the stopper by holding it at the receptacle, not at the shaft that fits into the chamber to avoid contamination.
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| *After the cleaning procedure store the chamber in 70% ethanol.
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| '''Cleaning with "Dead Cells":'''
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| *Every once in a while it might be necessary to wash the chambers with "dead cells" obtained from a e.g. former cell-experiment (frozen at -20°C). Inhibitors will be taken up by the 'dead cells' when filling the chamber up to the rim with a 'dead cell' suspension. The suspension should be left in the chambers with inserted stoppers for at least 30 min.
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| {{#set: Technical service =Chamber| Technical service =Contamination | Scientific service=Inhibitor }}
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| __SHOWFACTBOX__
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| {{Technical service}}
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