RE: bone IHC/large plus slides
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From: | "P. Emry" <emry@u.washington.edu> |
To: | Patsy.Ruegg@UCHSC.edu |
Reply-To: | |
Date: | Thu, 22 Jul 1999 16:55:20 -0700 (PDT) |
Content-Type: | TEXT/PLAIN; charset=US-ASCII |
Hi Patsy,
I saved this to get back to with questions.
I think I filled out one of the forms and sent it in. I'm
Patricia K. Emry. I could use some guidance in ihc and bone.
Would you see if I am registered and then send along the site for
ihc/bone? Thanks for your time.
I sectioned large specimens did H & E on one set and a lab guy did Brdu on
the other set. 3 out of 10 fell off the slides of the Brdu set, but all
my H&E's were fine. I washed and coated the slides with silane (?) as I
was instructed to do, but still lost a lot of tissue.
Is there another coating that can be tried or vendors that sell large
slide already coated?
My last question. What is HIER? I have seen other references to it, but
don't know what it is.
Thanks for your help.
Trisha
U of Washington, Seattle
On Thu, 3 Jun 1999 Patsy.Ruegg@UCHSC.edu wrote:
> Deb,
> Not much published specifically for bone, that I am aware of, we are
> pioneers in this area. We have been doing some ISH on frozen bone sections
> using the Instrumedics Tape Transfer method, just about impossible without
> it in my experience. Formic acid decalcified paraffin embedded zinc formal
> fixed is how I do IHC on bone samples, and if any pretreatments are required
> I use enzyme digestions rather than HIER. If you would like to join the NSH
> IHC Resource Group send me a fax number and I will fax you an application.
> The group has a general IHC reference list available to our members.
> Patsy Ruegg
>
> -----Original Message-----
> From: ebert.dc@pg.com [mailto:ebert.dc@pg.com]
> Sent: Wednesday, June 02, 1999 12:25 PM
> To: patsy.ruegg@uchsc.edu
> Subject: Re: bone IHC
>
> Patsy,
>
> I am just beginning to set up our lab for IHC and in situ on
> bone - frozen and
> paraffin. Do you have any recommendations for reading
> materials and any helpful
> hints? I'm sure there are many but a few to get me started
> would be helpful. I
> have never fixed, processed or cut bone frozen or in
> paraffin - only when
> embedded in MMA. This is quite a stressful thing to start
> when I don't have any
> experience. Thanks for any suggestions you may have.
>
> Deb Ebert
> Procter & Gamble Pharmaceuticals
> Mason, OH
>
>
>
> From: patsy.ruegg@uchsc.edu on 06/02/99 12:53 PM
>
> To: histonet@pathology.swmed.edu
> cc: (bcc: Deb Ebert-DC/PGI)
> Subject: bone IHC
>
>
>
>
> I missed the original thread on Bone IHC, so now I would
> like to add my two
> cents worth.
>
> I too have gone to fixing with Zinc formal which requires
> much less antigen
> retrieval, but when I do require pretreatment to access the
> antigen site, I
> use enzyme digestions rather than hier. We have found that
> we can control
> the process much better with enzyme digestion and it causes
> less damage and
> dislodging of the sections. I also use silane coated slides
> (plus slides).
> There are certain components in bony tissues that require
> enzyme digestion
> anyway and you must completely decalcify or the hydroxy
> appetite will block
> access to the antigen site, or take on everything because it
> is so porous
> and absorbent. Before IHC I always treat the deparaffinzed
> slides with 0.4
> M EDTA to assure complete decalcification. There are so
> many idiocyncrities
> about bone IHC, it is a whole different science from soft
> tissue IHC.
> Patsy Ruegg
>
>
>
>
>
>
>
>
Trisha
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