Does anyone have experience with neurons on sapphire discs especially with the coating protocol? We use Poly-D-Lysine - it´s great on glass, but neurons do not grow on sapphire.
Eric,
You probably have to do some carbon-coating of the discs. Then I would apply Poly-L-lysine, then place cells....
Cheers,
Thomas
Try 50ug/ml poly-ethylene imine (Fluka, Buchs,Switzerland).see Ruardij 2000 IEEE trans Biomed eng 47:1593-9 and Mouveroux 2001 Proc 23rd Annual Int Conf IEEE 745-748.
Hej Jean-Marc,
yes we managed to grow neurons on saphirre discs. We use Poly-D-Lysine (100 µg/ml in 0,15M Borate pH 8,4). 60000 cells were seeded per well (24 well plate) in neurobasal (600 µl). For the first 3 days grow the neurons without antibiotics. Let me know if it works!
Best,
Eric
Hi Eric,
I have a problem right after the growth while taking this saphirre discs out of the 24 well plate to be fixed in epon for electron microscopy. Did you faced this problem? If yes please let me know how did you managed as every time we try to take it out the neurons on saphirre disc are pulled out by the rest of the neuronal network on the well plate as we seed the whole well. Kindly reply as soon as possible.
Best,
Sneha
Hej Sneha!
I´m sorry but I have never observed such a problem! To take out the sapphire discs, I use a simple tweezers! Maybe you could sputter the discs with carbon following by Poly-L coating, to improve the attachment of the neurons to the sapphire. Another possibility would be to fix the cells on the discs with PFA/GA before you take out the discs. But depends what you want to do afterwards.
Hopefully you find a way to fix this problem!
Best,
Eric
Hi Eric,
Thank you for the response, I will try fixing them before taking out, just one more question did you use 24 well plate as well?
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