Re: [Histonet] Bone adhesion problems after HIER

From:Rene J Buesa

  I am somewhat confused about your e-mail, because it is titled "bone adhesion problems" and at the end it seems that you solved that problem, but are having problems with the IHC reaction.
  1-for the adhesion: I always used 1 mL of Elmer's glue in a regular 2L water bath and "fished" the sections with superfrost + slides (for both nails and bones). Did not lose sections during HIER.
  2-about the failure with IHC I cannot try to help you if I do not know what epitope your are looking for.
  René J.

"Leung, Patrick"  wrote:

I am having problems with cortical bone detaching from the slide during
the antigen retrieval step. The samples I am staining are paraffin
embedded mouse tibia.

I am currently using Superfrost+ slides. The AR is with citrate buffer
(pH6.1) from Dako performed in a waterbath for 15 minutes. The slides
are baked overnight at 59C prior to deparaffinizing. 

I have tried using a steamer and microwave, however I still ended up
with the same results. In addition Superfrost GOLD slides did not help
in keeping the bone on. I found that performing HIER at 70C for three
hrs as suggested in the archives to be ineffective in exposing my
antigen (although the bone did stay attached). 

Do you have any suggestions?


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