Hier and tissue architecture

[alexandrabetts]

I'm carrying out immunohistochemistry (several antibodies) on fibrous lesions. When using HIER I'm getting almost total loss of tissue architecture and tissue detachment from the slides both when using pressure cooker and microwave (citrate buffer pH6). Doubling the concentration of APES adhesive helped reduce loss of adhesion but there was still loss of architecture.
I've tried using citrate buffer pH 3 at 37C but that didn't work.
Any ideas?

Sandra

[richard03]

Sandra,

I am suggesting to use a little bit of gelatin in your water bath to solve the problem. If it didn't work, try to coat slides with gelatin solution

Recipe for Chrome Alum Gelatin Solution:
Gelatin, type A, 275 Bloom -------------------- 3 g
Chromium potassium sulfate ------------------ 0.5 g
Distilled water ----------------------------------- 1000 ml

Heat to 60 C to dissolve gelatin and then add chromium potassium sulfate.

Hope this helps.

Richard