I have a triple fluorescent staining (DAPI nuclei in blue, Ki67 nuclei staining in red and Mac3 staining membrane an citoplasm in green). I would like to quantify the amount of cells positive for Mac3 and Ki67. I managed to quantify cells positive for DAPI and Ki67 because those two staining colocalize, but I don’t know how to then screen for only cells Mac3 positive. Is there anyone who has an idea? I usually work with Fiji, but if Cellprofiler is a better option I can download it
I upload some images that I hope will help with understanding the problem.
Thank you very much,