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coatings for growing cells on glass coverslips - tissue culture and confocal microscopy (May/03/2005 )

Hi
I am trying to stain live cells grown on glass coverslips. My expiriments include a few rinses, and by the time i'm finished, most of my cells are gone! sad.gif
Does anyone have expirience with different coatings for the coverslips?
my cells are mesancephalic neurons (no need for trypsin-they are washed off by pipeting medium on them) and I am trying polylysine, fibronectin, goat serum, and laminin as coating.

-Rochelle-t-

Have you tried gelatin, collagen? Also if possible why not use chamber slides? Perhaps your cells may stick to those better instead and they are awfully handy.

-MaximinaNYC-

I have grown neuroblastoma cells on cover slips in the past. I found that simply leaving the cover slips in HCl for a few hours allowed the cells to stick better and stay on through staining.

-neuromatt-

QUOTE (MaximinaNYC @ May 4 2005, 05:46 AM)
Have you tried gelatin, collagen? Also if possible why not use chamber slides? Perhaps your cells may stick to those better instead and they are awfully handy.


Thanks,what is the protocol for colagen and gelatin?

-Rochelle-t-

Hey,
Try poly-lysine coated slides. If not you can try coating with poly-lysine yourself. Allow it to dry well.
But if you r trying for experiments which do not need live cells then just fix the cells using methanol or paraformaldehyde.
All the best

-ramakn-