Is Flow Cytometry Real?
Hey folks,
I've got cells expressing a fluorescent protein. Am getting a bizarre expression pattern I've never seen before:
Sample showing weird expression.
We want to quantify how many cells are expressing the fluorescent protein. I've done this hundreds of times. But I've never seen a treatment which causes an expression pattern like this, where the negative(?) population shifts so deeply into the positive gate.
See negative control, for reference:
Negative control with no mCherry.
Meanwhile, our positive control exemplifies the expression I'd expect to see:
Positive control showing typical expression.
Wondering the flow community has any advice? I've repeated this experiment a few times and it's all similar (I've deliberately chosen the most stark, though). Our cytometry techie took a quick look and found no issue. We're getting to the stage where the PI's are thinking up future research based off this finding, but something about the way this looks just isn't sitting right with me...
Any opinions appreciated,
King_of_Mints
P.S. There is technically some GFP expression in the sample as well, but it is incredibly insignificant (<0.5%)!