[Histonet] Re: bone implants

[Patsy Ruegg]

This is an age old problem, bone comes off easily especially during HIER,
which is why I have adapted all my bone IHC abs to use EIER instead of HIER,
usually with pepsin or proteinase K.  Also, it helps to heat the slides flat
on a slide warmer at 45dc overnight.  If all else fails I use elmer glue
coated slides but those can be problematic because the glue may react with
some ab reagents.

Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pruegg <@t> ihctech.net
web site www.ihctech.net
 

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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Reynolds,Donna M
Sent: Wednesday, February 15, 2012 9:49 AM
To: 'histonet <@t> lists.utsouthwestern.edu'
Subject: [Histonet] Re: bone implants

I don't know about implants but we were having a terrible time with bone
marrows coming off. Usually they came off during the HEIR using the steamer.
At that point we were placing our slides lying flat in a 55-60º oven
overnight which usually helps tremendously with difficult tissue. This was
followed with routine deparaffinization and hydration. It was suggested to
us to try a 70º water bath for 2-4 hours for HEIR. I was doubtful that my
antibodies would work but they did. In fact the staining was better with 2
hr HEIR than with 4. 

 Donna Reynolds Chief Histology laboratory, Research
Department Cancer Biology
U.T. M.D. Anderson Cancer Center, Houston, TX.
713-792-8106

Message: 4
Date: Mon, 13 Feb 2012 09:53:15 -0600
From: "del phillips" <dphillips <@t> vetmed.lsu.edu>
Subject: [Histonet] IHC and bone implants
To: <Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000a01ccea67$9958da90$cc0a8fb0$@vetmed.lsu.edu>
Content-Type: text/plain;	charset="us-ascii"

Does anyone have any suggestions for staining bone implants? I am having
trouble with wash offs. I use charged slides and they are still washing off.

 

Thanks

Del

 






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