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I wanted to do a few microscopy experiments with PC12 cells but when I plate them on glass they don't look very nice and there are hardly any cells attached. For growing them on plastic surface I coat the plastic with collagen. I tried coating class with collagen but the result was not great (still lots of floating cells). Any suggestions what kind of coating should I try? Maybe something else, like pre-coated dishes or maybe dishes from a particular manufacturer would be a better option?
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Vibor Laketa Tuesday, 29 October 2013 - 16:54 UTC
Edit: not as good for glass
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Vibor Laketa Tuesday, 29 October 2013 - 16:53 UTC
I did not try Cell Tek, I tried bunch of other extracellular matrix proteins (like fibronectin, gelatine etc.) and they all worked fine for plastic coating but as good for glass. If some one has tried Cell Ten please let me know...
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Alen Piljić Monday, 28 October 2013 - 17:02 UTC
OK... thanks Michael.
Vibor, did you ever try Cell-Tak? -
Michael Model Monday, 28 October 2013 - 13:29 UTC
There is a product call Cell-Tak from BD. But you may simply wait till they settle down and then take images if that works for you
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Alen Piljić Friday, 13 September 2013 - 13:34 UTC
Can you upload a protocol for this? :)
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Vibor Laketa Thursday, 12 September 2013 - 20:34 UTC
You should try poly-L-Lysine (PLL) coating. It works great for attaching PC12 cells to the glass (at least the ones in my lab... there are many different clones out there). The protocol is very simple, first you have to determine the optimal PLL concentration (that is the minimum concentration necessary for cell attachment, in my case that was 50uM). Than,
- cover the glass (coverslips or glass bottom dish) with PLL solution for 15min, room temp
- wash once with PBS
- wash twice with H2O
- cover the glass with PC12 growth medium for 5min (no cells, just the medium)
- seed the PC12 cells
- enjoy
good luck!