A phenomenon frequently forgotten is coincidence. If two cells enter the observation volume at the same time they will be seen as one event with the properties of both cells. This is a very common problem in my world (platelets) and should be considered as a possible explanation for any kind of unexpected dual expression of markers. It would be interesting to know if the frequency of CD4/CD8 doubles changes as the particle flow rate changes (i.e change the sample pressure or dilute the specimen.) Ken Ault
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