RE: [Histonet] Bone adhesion problems after HIER
This is a major problem in the land of mouse bone histology. The only
suggestion is to do as little HIER as humanly possible to get the
results you need. I have cut mine back to 10 minutes. Still get
beautiful immuno with minimal detachment.
>[mailto:email@example.com] On Behalf Of Leung,
>Sent: Thursday, January 11, 2007 12:18 PM
>Subject: [Histonet] Bone adhesion problems after HIER
>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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