I have images of tissue sections stained with three fluorescent dyes (DAPI and two cell markers). I need to count double and triple positive cells.
I started by dividing my multichannel image to single channels and was able to create binary mask of DAPI channel with nicely segmented nuclei (used gaussian blur, thresholding, LoG, and watersheding) and binary masks for my other channels (simple thresholding was just fine).
Now I would like to slightly grow my DAPI objects (by 1-2px), but without merging objects, and, by comparing the masks I have, create new masks for DAPI objects that are positive for one or both of my cell markers. Could anybody advice me on how to crate these new masks?
It would be perfect if I could have, in my new masks, only those DAPI objects that have more than few pixels in common with the other masks. As my images are from wide-field microscope I can sometimes see that I have two cells from which one is positive and the other is negative for my marker but they are most likely touching at some depth causing there to be a few bright pixels on my negative cell.
Thanks in advance.