Is anyone doing T17/Treg staining on liver biopsy specimens (paraffin fixed)?
I am getting a huge background.
Can I use ImageJ to quantify the degree of positivity I get, such that I could compare the relative infiltratino in two different samples?
For paraffin sections antibody selection and related epitope retrieval method are the most important steps.
For the background problem I recommend envision kit from Dako.
In my case, It was not possible to get reliable results for neutrophil counts in liver by ImageJ. In many case you will see unspecific staining of other cells/structures that make automated single cell counting impossible.
FoxP3 staining on frozen liver sections works fine (Dako envision with AP). Due to the minimal background, ImageJ could be used for determination of % Foxp3 stained area (of total area). However, I haven't tried paraffin sections with the clone FJK-16s antibody.
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