cleaning coverslip for protein/cell attachment

[ruchi jain]

Hi, Although the protocol looks quite standard and similar one works for me
but i quote my coverslips with polylysine at the end. In any case u can buy
precleaned coverslips from various companies. Just search for it!!! A quick
search gave me this link
http://www.sorvall.com/com/cda/landingpage/0,10255,2852,00.html
<http://www.sorvall.com/com/cda/landingpage/0,10255,2852,00.html>hope it
will b of use to you!!!

Best
Ruchi

On Thu, Jul 16, 2009 at 6:41 AM, guanyuan fu <guanyuanfu from gmail.com> wrote:

> 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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>



-- 
Ruchi Jain, Ph.D.
Postdoc Associate
Internal Medicine-Infectious Diseases
Yale University
ruchi.jain from yale.edu
203-737-1197