I am new in this forum and I was wondering if you could help me a little bit.
My aim is to quantify the fluorescent signal (nuclear and cytoplasmic) in treated and untreated samples.
After subtracting background and setting the threshold, I tried 3 different ways:
- I just did ‘Measure’ and I got for example:
Area= 1240.45; Mean=13.494; IntDen=16738.21
- I did Analize Particles (size and circularity=0 for this example) and I got:
Area= 1216.175; Mean=6.552; IntDen=69.696
- I applied the Threshold and converted to a mask and I got:
Area= 1216.175; Mean=254.925; IntDen=1318.851
Which one would be the best approach and, I know that the IntDen= Area*Mean, but why the IntDen is so different in the 3 cases?
Looking forward to your reply.
Thanks a lot!