Manual vs. Auto compensation

Hope all are having a great weekend and not in the lab on a Saturday night.

I'm a relatively knew guy to the sort/flow world and need to set some things
straight so I hope those with more experience can give me their feedback.

I have people in the lab who complain about me not doing manual
compensations and instead have the Aria calculate them for me. And when I
have the Aria compensate, they want to manually compensate after they record
the data because the plots "don't look right." I mean, isn't that simply
producing "make-belief" data? Or am I missing something? I'm not trying to
criticize those who do manual comp. I just want to understand why do I need
to manually compensate when it's all based on mathematics and can be done by
the machine?

And the frustrating part is that when they bring me the compensation
samples, they contain cells that are different from the ones in their
analysis samples. I try to tell them that the autofluorescence of both cell
types is most probably different and that might be the reason why some of
the compensations don't turn out correctly. But they dismiss my suspicion,
claiming that it should not affect the compensation since we're dealing with
colors and not cell density. Is that true?

I'm stressed out. I need some ice cream.

-- 
Nidal Muvarak
Abramson Research Center
Children's Hospital of Philadelphia
3615 Civic Center Blvd
Philadelphia, PA 19104