I think I posted this in the wrong place. So I copied in here.
I labeled my cells with DAPI and phalloidin to identify nuclei and outline of individual cells, respectively. Although I can clearly see single cell outlines in my fluorescent images, “identify secondary objects” module doesn’t follow true outlines but mostly find non-specific particles.
What am I doing wrong? Can you help me?
P.S.: I attached my DAPI and phalloidin stained images. Could you help me finding individual cell outlines?