🚀 go-pugleaf

Dear Steffen,

I use clear Wet 'n Wild.  It is toluene/formaldehyde free, I wonder whether this is what makes a difference.

Clonetech recommends rubber cement or agarose for sealing, but we found no reason to not use nailpolish, our standard sealant.



Anda Cornea, Ph.D.
Oregon Regional Primate Research Center
505 NW 185th Avenue
Beaverton, OR 97006
ph: (503) 690-5293
fax:(503) 690-5384


>>> Jacques Paysan <paysan@UNI-HOHENHEIM.DE> 03/10 2:42 PM >>>
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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