cleaning coverslip for protein/cell attachment

guanyuan fu via methods%40net.bio.net (by guanyuanfu from gmail.com)
Wed Jul 15 20:11:57 EST 2009


Hi, all:

I have some trouble with cleansing my coverslips for protein coating. The
coverslips (22*22 mm) seemed to be very hydrophobic, as the protein solution
(500 microliter) tended to aggregate into a big drop instead of disperse all
over the surface of the coverslip. Normally, about 100 microliter solution
should be able to cover the whole surface. I don't have a way to measure how
much the protein is attached, but it seems to be very few if any based on
the fact that my cells expressing activated receptor for that protein could
not attach to the surface.

The procedure is listed below:

1) Wash the coverslips with chromic sulfuric acid (fisherbrand cleaning
solution) for overnight.
2) Rinse with running tape water for 3 hrs to remove chemical residues from
step 1.
3) Rinse with distilled water 3 times each 30 mins.
4) Wash coverslips with 100% alcohol (mixed alcohol, not pure ethanol) for 2
hrs
5) Discard alcohol, and separate coverslips one by one to let it air dry.
6) After dry, store in glass plate then autoclave.

Please give me any suggestion on modifying the above procedures since i am
new to this methods. or if you have a good substitute (precleaned coverslip
for protein coating to study cell spreading on the coated protein only),
please also tell me where i can buy it. Thanks,

Guanyuan Fu


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