I’m trying to quantify RNAscope data. I have three probes and DAPI(C4). I have identified nuclei and spots.
One of my probes is highly dendritic , why it can be found every where(image C3).
If I put a pseudo nucleus in the image, I will get as many spots as I have for a true nucleus.
If I put a threshold for C3=30 spots, how can I be sure those spots is nuclear and not dendritic.?