I tried as your suggestions and I obtained this excel data:
In same case, I obtained very high values (as you can see in the highlighted line): how can interpret them? At the end of this analysis I want to represent every single object in a graph with a scale from 0 (negative 488 object) to 1 (100% 488 positive) , where the 0 object are the negative for 488, 0.5 an object with a average positivity, etc..so I need also the 488_positive intermediate values.
I need also some suggestions to put a threshold to define the 488-negative objects ( I obtained only 2 objects that are 0.0, but a lot of them had a 0.000XXX value, in fact the 488 positive objects are 101 on 113 total Dapi objects).Foxg1_488.cpproj
Thanks for your helpful advices.