This is another question about ambiguity and the question itself may be ambiguous: What can one do with a stain in which the whole population shifts into a higher fluorescence and no discernible separation of a subpopulation ocurrs? (The population leaves a gap in the left hand side) Say, for example in the case of activation markers CD69 and CD25. After activation all the cells are on their way of expressing it, but after short term incubation only a percentage will shift away from the "background fluorescence", "isotype fluorescence" or "negative" quadrant. Can the change in MFI for the whole population be considered relevant (if indeed they are all positive?), or is the only right way to look at this by cutting off the "negative" and looking at the MFI of the "positive" population? Could a possible explanation be that cells who were initially in the low "negative" population have actually shifted into a higher "positive" fluorescence, but this shift is being masked by the background fluorescence? Any answer and /or lengthy explanations would be greatly appreciated. Thank You. Artur -- ÐÏࡱ
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