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Tony Hodge tph at mrc-lmb.cam.ac.uk
Fri Feb 5 07:43:49 EST 1999


Louise Neville <louise.neville at MAIL.CHMEDS.AC.NZ> wrote:

> Hi all
> 
> I have just found this site and hope someone can help me!  I have grown and
> transfected cos-7 cells on coverslips with EGFP variants.  I now want to
> fix and mount these onto slides to visualise fluorescence.  The clontech
> booklet suggests mounting with PBS and sealing with molten agarose or
> rubber cement, I would rather use a mounting agent as the slides seemed to
> dry out using agarose.  Does anyone know of an appropriate mounting reagent
> which doesn't interfere with GFP fluorescence.  I would really aoppreciate
> any suggestions
> 
> thanks in advance
> 
> Louise Neville

I have used Mowiol (essentially Polyvynyl alcohol) as a mounting agent.
Only drawback is it take a while to dry out.  A drop on a slide with the
(rinsed in PBS) coverslip inverted onto it is great for up to a week.

2.4g Mowiol (Hoechst) or Gelvatol/Gelutol (Monsanto), 6g Glycerol.
Stirr to mix.
Add 6ml water and leave for several hours at room temperature.
Add 12ml 0.2M  (or 0.1M) Tris pH8.5, heat to 50C for 10 minutes with
occasional mixing.
Once dissolved clarify by centrifugation at 5000 x g for 15 minutes

Aliquot and store sealed at 4C (or may be frozen).

tony

-- 
Tony P Hodge
tph@*nospam*mrc-lmb.cam.ac.uk  (remove *nospam* )
http://131.111.84.120/tph/
http://131.111.84.120/tph/psion/



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