[Histonet] RE: mouse brain fixation Attn: Annette Featherstone

[gayle callis]

When you say zinc fixative, which one are you talking about?  Becksteads
Zinc TRIS buffer made up with zinc salts and formalin free?  Or zinc
formalin.  The Becksteads zinc salt fixative does work, as cited in =
several
papers in the literature, and we tried it but preferred to use frozen
sections, just faster overall.   And if you overexpose the tissues to
dehydrant, clearing agents and heat of paraffin, the sectioning can be =
worse
with the Beckstead fixative.  It makes the tissue dry and crunchy.  IF =
you
do not fix the tissues properly and totally, then the alcohols used in
processing  will finish the fixation, and that isn't going to contribute =
to
good sectioning either.  How you make it up is critical, and should be =
done
according to the original publication or Nitta et al paper. =20

=20

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Gayle M. Callis  =20

=20

From: TF [mailto:tifei <@t> foxmail.com]=20
Sent: Tuesday, June 30, 2009 11:33 PM
To: gayle callis; 'Histonet'
Subject: Re: [Histonet] RE: mouse brain fixation Attn: Annette =
Featherstone

=20

I tried Zinc fixative I made myself.

Does not work at all? And, the tissue quality is very bad.

Any comments?

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2009-07-01=20

  _____ =20

TF=20

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=B7=A2=BC=FE=C8=CB=A3=BA gayle callis=20

=B7=A2=CB=CD=CA=B1=BC=E4=A3=BA 2009-06-30  23:57:35=20

=CA=D5=BC=FE=C8=CB=A3=BA 'Histonet'=20

=B3=AD=CB=CD=A3=BA=20

=D6=F7=CC=E2=A3=BA [Histonet] RE: mouse brain fixation Attn: Annette =
Featherstone=20

Annette,=20

=20

You did not say what immunos e.g. antigens you are trying to stain?   =
Nor

how you are doing the actual staining method?  More information would =
help

please.=20

=20

If you are trying to stain for murine CD markers or some other cellular

antigens, there are not many that work after FFPE.  If you need to know =
this

information, go to SEROTEC and look at murine CD or other cellular =
markers,

and see if the antibody will work with FFPE paraffin applications.   BD

Bioscience also has applications that work, tested, reported, IHC or =
frozen

sections and found in their Technical Data Sheets for any given =
antibody. =20

There are some other fixatives to try for CD and other cellular markers =
-

SEROTEC often refers to PLP (paraformaldehyde-lysine-periodate with =
recipe

found on web through IHCWorld or Immunoportal, Google the keywords) =
while BD

Bioscience refers to Becksteads Formalin free Zn TRIS buffer fixative. =
If

you need the recipe for that, I will be happy to send.  Otherwise BD

Bioscience would be happy to sell that to you but it is cheaper and easy =
to

make up in the lab.=20

=20

Gayle M. Callis

HTL(ASCP)HT,MT

Bozeman MT=20

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 =20

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You wrote: =20

=20

We have been working with mouse brains that were fixed in 10% formalin =
for

one month. We are currently getting "no" staining with our immunos. Can =
this

really be the problem in light of antigen retrieval methods?

=20

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=20

Annette Featherstone

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=20

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