First one is singlets from Wild-Type mouse (bone marrow):
Next is singlets from GFP+ mouse (bone marrow):
The bone marrow cells from GFP mice have a lot of fluorescence also in the PE (575nm) and PI (610nm) channels; PI isn't so important for our experiment (thus why the PI voltage probably isn't optimal), but PE is...another marker uses PE as a label:
For comparison, the wild-type cells:
I compensated the FITC and PE channels, thinking this was the optimal/best I could do given the circumstances. The assumption was that all of the cells are positive, and thus the cells centered around 10^2 are positives. (FlowJo warns me that 6% of the cells are on the chart edges):
I compensated the FITC and PE channels, thinking this was the optimal/best I could do given the circumstances. The assumption was that all of the cells are positive, and thus the cells centered around 10^2 are positives. (FlowJo warns me that 6% of the cells are on the chart edges):
I suppose I could compensate MORE? Maybe the cells centered around 10^2 are negative (different autofluorescence than the wild-type?), and the ones between 10^3-10^4 are the positives? If I do, now 58% of the cells are on the chart edges! And I still get a very weird vertical line at the right-hand side.
