Good morning and thank you for putting to everyone’s reach such a great software!
I have gotten a lot of very helpful tips from previous blog entries but I still have a couple of pending questions. I am trying to quantify cytoplasmic vs. nuclear staining and am attaching sample pictures as well as the project I have designed.
a) The lymphocytes I am analysing tend to have shapes that are not always circular and I have found that using propagation as means to identify secondary objects yields better results then Distance - B (that most times disregards any cytoplasm that is not circular). Are there any caveats to this approach?
b) In the attached example, the cytoplasmic outline in blue sometimes fills up the whole cytoplasm (for e.g. rectangular looking cell in the bottom left), and in other instances the outline does not fill the cytoplasm but just outlines it. It is really important for me to be able to accurately quantify the cytoplasmic intensity. Will such 2 cells be evaluated differently by the software? Which outlining should I be aiming for? Would you suggest doing anything else differently?
Many thanks in advance for your support!
IFNg.cpproj (951.1 KB)