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Flow Cytometry Inconsistent Between Days - Mitotracker Green FM Staining

flow cytometry mitochondria

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#1 aegreen

aegreen

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Posted 06 July 2013 - 03:30 PM

I'm using Mitotracker Green FM to measure mitochondrial mass on cells on separate days. Following staining, I run the cells through our FACSCalibur with the same settings every.

The problem is that my my geometric mean varies from 50-200 depending on the day for the same cell line and treatment. I run an unstained sample and it is fairly consistent plus or minus ~3. Immediately this makes me think there is variance in my staining. I aliquot my stain and never reuse aliquots. My staining concentration and time is the same. No matter what I still have a lot of variance.

Is it possible that this variability is from the flow cytometer PMTs? Other than using calibration beads is there anything I can do to correct for these day to day variances?

Any help would be apprecaited





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