|
|
| (6 intermediate revisions by 3 users not shown) |
| Line 1: |
Line 1: |
| ==Problem==
| | #REDIRECT [[MiPNet19.03_O2k-cleaning_and_ISS]] |
| | |
| Lasting effect of inhibitors (i.e. rotenone, antimycin A), as observed by persistent low fluxes, when working with permeabilized fibers.
| |
|
| |
| | |
| ==Cleaning the Chamber==
| |
| | |
| | |
| '''Clean the chamber after an experiment involving lipid-soluble inhibitors (such as oligomycin, rotenone, or antimycin A):'''
| |
| | |
| *The chamber must be cleaned rigorously with ethanol, since such inhibitor(s) are difficult to be washed out from the chamber and may inhibit mitochondrial respiration in subsequent experiments.
| |
| *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.
| |
| *Also rinse the surface and cannula of the stopper with distilled water properly.
| |
| *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.
| |
| *Remove the stopper and siphon off the ethanol to empty the chamber. Repeat these cleaning steps with 70 % ethanol three times.
| |
| *Then fill the chamber with absolute ethanol (99.6 %) and insert the stopper making sure that the ethanol fills up the receptacle. *Replace the perspex cover on top of the stopper and leave for 15-20 min.
| |
| *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.
| |
| *Siphon off the ethanol from the chamber and rinse it with distilled water three times.
| |
| *Rinse the stopper by holding it at the receptacle, not at the shaft that fits into the chamber to avoid contamination.
| |
| | |
| | |
| {{#set: Scientific service= Inhibitor|Scientific service=Chamber cleaning}}
| |
| | |
| {{Scientific service}}
| |
| | |
| | |
| __SHOWFACTBOX__
| |
| {{Troubleshooting}}
| |
