[Histonet] paraformaldehyde autofluorescence
gayle.callis <@t> bresnan.net
Wed Mar 19 09:49:33 CDT 2008
It is not the heat used to dissolve paraformaldehyde that causes
autofluorescence, it is caused by aldehyde induced autofluorescence when the
PFA crosslinks proteins. There are places to buy PFA in liquid form to
avoid having to weigh and dissolve. We never bother to add NaOH if we heat,
and we also buy our paraformaldehyde from Sigma.
We always heat our PFA but never exceeding 60C. It will go into solution
around 56C, with stirring in approx 15 minutes or so.
I suggest you (and the researcher) read about on autofluorescence on the
IHCWorld website. There is a superb discussion on both fluorescence and
----- Original Message -----
From: "Helen Beard" <hbeard27 <@t> hotmail.com>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Tuesday, March 18, 2008 6:45 PM
Subject: [Histonet] paraformaldehyde autofluorescence
We have been stirring pfa overnight @RT to dissolve, avoiding heat, as one
of our researchers believes heating to 60-65C causes autofluorescence in
tissue. Has anyone had this problem with autofluorescence? I would prefer
to heat and add NaOH. Also what are thoughts on using fresh, frozen 4%pfa?
Thanks in advance
Adelaide South Australia
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