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How to coat coverslips before seeding cells?



jennycharlies Posted 18 August 2011 - 06:55 AM
Hi,

I am going to seed my cells onto coverslips inside the wells of a 12 well plate and was wondering what is the best way to coat the coverslips?

I have 18mm coverslips and need to coat with poly-D-lysine. But presumably I need to sterilise the coverslips first.. What is the best way to go about this? I assume I need to coat the slides while in the wells of the plate?

Thanks for your advice!!

Chakchel Posted 18 August 2011 - 11:10 PM

Hi!

Yes, you first should sterilize your coverslips. You can do this either by putting them into the autoclave, e.g. wrap several in a little "bag" of aluminium foil, or by dipping them in Ethanol and shortly flame them (too long flaming will let them burst).

Then you can directly cover them in your well. Put your poly-D-lysine in the well, so that the coverslip is covered, but not too much, then let it dry in the hood for several hours. If then there is still some liquid, you can suck it of.
Finally wash the wells once or twice with PBS.

Done, and ready to use.

Edited by Chakchel, 18 August 2011 - 11:11 PM.

fysio lab Posted 19 August 2011 - 02:25 AM
Hello,

we put non-sterile coverslips in a glass petridish, wrap that in alu-foil, autoclave it and before use we put the CS in the wells with sterile forceps. After that we put our coating solution on the CS (in our case 2% gelatin), we leave it 30' in the incubator. Then we remove the gelatin and let it dry for a few minutes in the flow, lid open... meanwhile we split and count our cells. All together this doesn't take much time.
Good luck

1 Comments

I use cover-slips for my primary neuronal cultures and I DO NOT STERILIZE them at all. I open the package under the hood, I put them into the wells using my fingers (wearing gloves, EtOH spray first) and then after I add the poly-D-Lysine + Laminin solution in PBS. Just before platting the cells I suck off the solution and that is all. I had not any contamination for the last 5 years.
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