RE: bone IHC/large plus slides

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From:"P. Emry" <>
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.

U of Washington, Seattle

On Thu, 3 Jun 1999 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: []
> 		Sent:	Wednesday, June 02, 1999 12:25 PM
> 		To:
> 		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: on 06/02/99 12:53 PM
> 		To:
> 		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


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