Dear all,

I am working with melanoma cryosections from minipigs. My aim is to characterize tumor-infiltrating immune cells between different age groups, mainly by immunohistochemistry. I do multiplex indirect immunofluorescence staining with primary antibody e.g., CD4 and CD8, then incubation with secondary antibody e.g., Alexa 488 and Alexa 555, and of course DNA staining with DAPI. For image acquisition I am using Leica SP5 confocal laser microscopy, then for image analysis I am using imageJ software. My question is how to quantify double positive signals using imageJ? Is there an easy way to do that? I searched in the internet for solutions but they got me very confused. Is there anyone who is experienced and can probably assist me with this please?

Thank you so much in advance!