Hi, everyone. I have been having some problems setting a pipeline so I hope you can help me. I need to quantify a protein in different cellular compartments (cytoplasm, nucleus and nucleolus) and meassure different variables and then export it to an excel.I have special interest in measure colocalization and intensity on nucleai on my protein of interest, and also for the nucleolus marker. I used IF microscopy and the nucleolus was stained with fibrillarin (nucleolus marker (red chanel)). The pipeline I am trying to build should have a way to identify nucleolus, nucleous and cytoplasm and be able to quantify my protein of interest in those compartments and excluding the others. This is what I have so far, but I think, I might have excesive modules or it might have some mistakes. I forget to add, that I am particularly interested in measure the area and shape of the nucleolus and nucleus (note, that fibrillarin does not stain completely the nucleolus therefore I used DAPI low intensities as predictor of nuclolus).
I will 20.12.17 nucleolo and protein interest.cppipe (13.7 KB)
Thanks in advance