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Dear Steffen

I found some melted parafin most useful to replace nail polish (I picked
this up from a newsgroup discussion some time ago).  Simply apply with the
edge of a heated slide that you dip into a block of parafin.  With a little
practice this works very easily. No effect on GFP and other fluorescent dyes
at all.

Jacques


----- Original Message -----
Sent: Friday, March 10, 2000 8:28 PM
Subject: Nailpolish. was: Losing GFP Signal


> Anda,
>
> wheras I never had problems with overnight fixation of cells in 1.6 %
> formaldehyde, I did have problems with the nailpolish: I have tried
several
> ones and they all killed the GFP before the slide was even on the scope. I
have
> heard rumors of nailpolish that does not harm GFP, but was never able to
hunt
> one down, therefore PLEASE:
> Could you let us know the exact brand name, product number or whatever is
> usefull to find the nailpolish you are using?
>
> Thanks!  Steffen
>
> --- Anda Cornea <corneaan@OHSU.EDU> schrieb:
> >
> > >>> "Goodhouse, Joseph" Long
> > For fixation employ low percentage paraformaldehyde for the shortest
periods
> > of fixation as possible( 5- 10 minutes for cells).  Samples should be
> > mounted in glycerol based mounting medium.  I have found 90% buffered
> > glycerol to be best.  Avoid using nail polish as sealant of coverslips.
In
> > a short time this will wipe out your GFP signal.
> >
> > We routinely use 4%paraformaldehyde, 30minutes fixation, for cells
expressing
> > a GFP-membrane receptor chimera and seal slides with nailpolish.  We see
no
> > loss of intensity when compared with live cells expressing the same
> > construct, not even after a few months storage.
> >
>
> =====
> Dr. Steffen Dietzel
> Dept. of Cell and Structural Biology
> University of Illinois at Urbana-Champaign
> 601 S. Goodwin Ave.
> Urbana, IL 61801
> Phone: +1/217/333-8372  Fax: +1/217/244-1648
> e-mail: dietzels@yahoo.de  Web:  http://www.life.uiuc.edu/belmont/dietzel
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