Re: bone marrow trephines

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From:Mick Rentsch <ausbio@nex.com.au> (by way of histonet)
To:histonet <histonet@magicnet.net>
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 Dear Marjorie,
we've been using FormalSaline/Formic Acid for more years than I can remember
on all our trephines. At the time of Aspiration we put the trephine directly
into this Fixative/Decal for a minumum of four hours, but more often then
not it's about 8 -10 hours as we normally do our own trephines at about
0800hrs. At four hours decalcifying is close to complete, and even though
sometimes a bit "crispy" you can still get good sections. If you leave the
trephines overnight in this Fixative/Decal, there doesn't appear to be much
deterioration and detail is still excellent.
It may well be that any acid decalcifying agent will be adverse for your
studies including Trichloracetic Acid, while I've only used EDTA on rare
occasions and then only to ensure the best possible detail, I only did so
for PSA and AcP, which were excellent but required two days to decalcify the
trephines.
The recipe I use for  5 LitresFormal Saline/Formic Acid is as follows:-
Sodium Chloride Analar 42.5g
Formaldehyde 38/40% Analar 500mls
Formic Acid 90-95% Analar 1600mls
Make up to 5 litres with RO Water >4MOhm.
Label as UN2810 Corrosive Liquid N.O.S. Decal Reagent. Give a two year
expiry and pack in 1 Litre DGA approv'd bottles.
Decant 35mls/ 70ml specimen jar labelled expressly for Bone Marrow
Trephines.
Hope this is of some help, Regards
Mike Rentsch (Downunder)

-----Original Message-----
From: Hagerty, Marjorie A. <mhagerty@emc.org>
To: Histonet <histonet@pathology.swmed.edu>
Date: Friday, 13 November 1998 8:29
Subject: bone marrow trephines


>
>Histonetters,
>We are currently using RDO for one hour to decal our BM core bxs. This
works
>perfectly for us, they are easy to cut, and the nuclear detail is
excellent.
>I think, however, that the HCL in the decalcifier may be interfering with
>our Kappy/Lambda immunoperoxidase staining. I tried an antigen retrieval
>solution specifically for decalcified tissue but that did not seem to
>correct the problem. Doing this on one case does not count as any kind of
>"study" so this may be an excellent product. We would like to try a non-HCL
>decalcifier. Alex metioned formal citrate but our clinicians would not
>tolerate the length of time it takes. Does anyone know of a decalcifier
that
>takes no more than an hour for very small core bxs and does not use HCL?
>Thanks
>Marg
>EMC, Rancho Mirage, CA
>




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