RE: B5 fixative and immunos

From:"Drew Sally A." <>

I'm not sure what the innovative method was, but I can tell you what we do
with our B5-fixed materials.  For B5-fixed LN we would do 2.5 minutes in a
pressure cooker with 1mM EDTA(disodium) pH8.0.  For our B5-fixed decal'd
bone marrows we use DeCal Retrieval Solution(following BioGenex's protocol)
followed by 2.5 minutes in the pressure cooker with 1mM EDTA.  We run the
monoclonal CD3 from BioGenex  on our Ventana ES/genII with an amplification

Sally Ann Drew-MT(ASCP)
Univ. of Wis. Hospital and Clinics
IHC/ISH Clinical/Research Lab
Madison, WI 

> -----Original Message-----
> From:	Donna Sitrin []
> Sent:	Wednesday, October 11, 2000 6:33 PM
> To:
> Subject:	B5 fixative and immunos
> Hi all!
> Somewhere during the symposium I caught wind of an innovative retrieval
> method or treatment for B5 blocks to improve antigenicity. From time to
> time, this same problem surfaces in my lab. Once again, I am having a
> problem with CD3 staining on lymph nodes and bone marrow cores fixed in
> B5,
> and I know there was a lot of information out there in regard to this in
> particular.
> I was hoping to tap your memories and that they are functioning much
> better
> than mine.  If anyone has any information on CD3 staining on lymph nodes
> and
> decalcified bone marrow cores fixed in B5, please jog my memory.
> All help will be greatly appreciated!  (I can't talk the pathologist out
> of
> B5 unfortunately.
> Thanks in advance for all help,
> Donna Sitrin

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