I'm examining double-labeling using cFos (nuclear) and oxytocin (cytoplasmic) and I have found that cells that appear to be co-labeled only have cFos around the surface of the nucleus, while the single-labeled Fos cells have clear and normal looking (dense, circular) labeling. I've attached a picture with cFos in green and oxytocin in red.
I can't find anything on this. Are these weirdo cells actually cFos positive? Or is it impossible to call a cell cFos+ if it isn't densely filled?
Although not in the specific case, I have seen this before when perfusion-fixation is not very good and large neurons get fixed unevenly, with the periphery of the nucleus always better preserved than the core. If I were a reviewer I won´t accept this as a definitive proof of cfos expression in these cells.
I did that - it's definitely just sitting on the outside of the nucleus. You can see the outside of the ball and when you get the "completely focused" image the center just looks spotty and sparse.
I tried a fresh antibody in 4 other rats and haven't seen it again. I think you two are right, bad fixation or something similar.
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