CONFOCALMICROSCOPY Archives

March 2000

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Subject:
From:
Robert Palmer <[log in to unmask]>
Reply To:
Confocal Microscopy List <[log in to unmask]>
Date:
Fri, 10 Mar 2000 16:24:21 +0000
Content-Type:
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Well we are certainly into the realm of voodoo now.  I am now using Almay
Creme (hypo-allergenic - maybe that's the secret!).  The shade is Canyon.
this deos not kill GFP in Streptococcus gordonii or Mycobacterium
smegmatis.  I actually prefer Bloodorange however.  It goes better with my
accesories...
Rob

>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 <[log in to unmask]> 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: [log in to unmask]  Web:  http://www.life.uiuc.edu/belmont/dietzel
>__________________________________________________________________
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