[Histonet] Re: Paraformaldehyde use question for geniuses
Gayle Callis
gayle.callis <@t> bresnan.net
Thu Apr 24 17:02:27 CDT 2008
Dear Teri and Fellow Histonetters,
Teri's words of wisdom on fixation and storage of samples are well taken.
Please provide the vendor for the 16% paraformaldehyde in ampules so our
FACS technician doesn't have to weigh out PFA powder anymore. She is going
to be delighted.
Gayle M. Callis
----- Original Message -----
From: "Johnson, Teri" <TJJ <@t> Stowers-Institute.org>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Thursday, April 24, 2008 2:55 PM
Subject: [Histonet] Re: Paraformaldehyde use question for geniuses
Geoff,
Have you performed experiments that show more of a benefit for storing
samples in formalin over 70% alcohol in your lab? I have one researcher who,
based on their experience in cultured cells, knows that anything over 10
minutes of fixation time causes their antibody staining to diminish using
one particular antibody. I'm well aware of the current trend towards longer
fixation times and generally believe that more is better. Because of
evidence to the contrary, I cannot bring myself to recommend as a matter of
routine, to store samples in formalin, especially for research purposes.
I much prefer to have the samples paraffin processed or cryo embedded after
a defined period of time in fixative, and store the tissues in those two
forms rather than anything aqueous or alcoholic.
Jackie - I'm not aware of any vendors who sell 4% formalin made from PFA in
large quantities, simply due to the problem of re-polymerization upon
storage. We get 16% solution in ampules, and then dilute in PBS to 4%.
Perhaps that might work for you?
Teri Johnson, HT(ASCP)QIHC
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, MO 64110
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