I am writing to request recommendations on positive detection parameters in the analysis of the immunohistochemical marker with DAB of CD68. We had problems with positive cytoplasmic detections.
Thank you very much for your help.
Macrophages are particularly problematic to segment, and I usually recommend using some kind of area detection rather than counting. There is no guarantee that most of the staining you see is from cells with nuclei within the plane of your tissue slice.
There are a couple of other posts on macrophages around that might give you some ideas, but for anything more specific, I would recommend providing examples of exactly what is going wrong.
Mildly applicable, maybe: QuPath-Accurate cytoplasmic stain measurements
I agree with @Research_Associate. Macrophages are difficult to segment, specially because they vary a lot in terms of size, shape, they may have several nucleus, they can form clusters…
So in general, their sdensity is expressed as a percentage of stained tissue, rather than a number of cells per unit of tissue.
Did you ever find a solution to your problem? We are trying to something similar with CD68 and we would like to hear how you tackled this particular situation. Did you switch to another program or did you manage to use QuPath to your satisfaction?