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| == Problem ==
| | #REDIRECT [[MitoPedia:_Respiratory_control_ratios]] |
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| At a high mitochondrial concentration (0.5 mg protein/ml), ADP is exhausted very rapidly. The slope increases sharply and returns to State 4 (LEAK state), even before the plot of flux shows the true maximum value of OXPHOS capacity (State 3), resulting in a low apparent RCR.
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| == Tests and Solutions ==
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| # Reduce the data recording interval (standard is 2 seconds) to the minimum of 0.2 s (setting in the Oxygraph Control window; see [[MiPNet12.06 O2k-Start]]). As the data recording interval is reduced, the flux appears more noisy, but represents transitions more accurately and reduces the apparent time-delay. At high flux per volume, the increased noise level presents no problem. Flux then shows a period of constant state 3 respiration rather than a sharp peak, and routine analysis is possible in DatLab 4. | |
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| # Reduce the mitochondrial concentration, thus prolonging the duration of state 3 respiration over 120 seconds. If flux reaches a constant value at state 3, then the maximum value is represented accurately by the plot of flux, and routine analysis is possible in DatLab 4.
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| # Export the data into [http://www.oroboros.at/index.php?id=o2k-datlab2 DatLab 2], and apply a time correction (signal deconvolution), as described by [[Gnaiger 2001 RespPhysiol]].
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| # See also ‘Notes on [[Time resolution]]’.
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| {{#set:Scientific service=Isolated Mitochondria | Scientific service=Respiratory States}}
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| __SHOWFACTBOX__
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| {{Troubleshooting}}
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