Re: Losing GFP Signal

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#46

From: Anda Cornea
Date: Fri, 10 Mar 2000 00:00

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


>>> "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.

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From: Anda Cornea <corneaan@OHSU.EDU>
Subject: Re: Losing GFP Signal
Date: 2000/03/10
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