[Histonet] Bone IHC

[Rene J Buesa]

If you have a bone that will require IHC, you will have to make sure that the Gross section is no more than 1.5 mm thick and decalcify it in EDTA. The you have to make sure that the infiltration is complete. These are the 2 initial factors.
Later you will have to section it as thin as you can, place the sections in (+) slides let them drain completely before going into the oven. Extra time in the oven is not really required if the aforementioned steps are done.
Then add enough dishwasher soap to the HIER solution to get a solution of 2% and you will dewax and retrieve the antigen simultaneously. The sections will survive to complete the IHC procedure.René J.

--- On Thu, 10/7/10, Vanessa J. Phelan <vjp2105 <@t> columbia.edu> wrote:



From: Vanessa J. Phelan <vjp2105 <@t> columbia.edu>
Subject: [Histonet] Bone IHC
To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu>
Date: Thursday, October 7, 2010, 3:13 PM


Hi everyone,

I was wondering if anyone has any tricks on how to get bone sections to stop lifting off the slide through the IHC process? I leave them in the oven for quite a while to make sure they are baked on, however after antigen retrieval  (pressure cooker for 20mins) most of the boney part of the tissue comes off and the marrow and muscle stays put! The sections are cut onto superfrost plus slides.

Any help would be much appreciated, thanks.

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