VSquared

I would try giving the cells an extra wash in fixative, maybe two more, but I wouldn't do more than 3-5 total washes. That should alleviate the cytoplasm problem. As for the chromatids splitting, I'm not sure. We sometimes have that happen too, but I'm only a "year old" in this lab and I'm not sure why that happens yet. I know your question is from a while ago so I'm betting you already fixed your problem...?