General Research Topics > Cell Biology

Collagen background with cell monolayers

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phergert:
Ok, so I have a PI that grows his fibroblasts  on collagen (imperative to his experiment) and the back ground behind the cells of the collagen is driving me nuts. He is looking of course for nuc vs cyto protein location, but dang the images are awful.  I use a methanol fixation and do  simple IF protocol and it turns out beautifully without cells growing on collagen.  I am wondering if anyone has any suggestions so I can produce a publishable image?  I used to use NaBH4 with tissue to help with crosslinking,  and it might just be impossible, but I thought I'd ask.  I've been doing serial EM mostly until I relocated back home and have been out of the IF game for a while, I thought someone might share an idea?  Thank you very much...

MT Scientist:
What are you staining for?

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