[Histonet] Problem with liver fixation

[Rene J Buesa]

Bouin's will not do better than what you describe. Are you using antigen retrieval?
René J.

--- On Sun, 9/5/10, Itai Moshe <itai.moshe <@t> mail.huji.ac.il> wrote:


From: Itai Moshe <itai.moshe <@t> mail.huji.ac.il>
Subject: [Histonet] Problem with liver fixation
To: histonet <@t> lists.utsouthwestern.edu
Date: Sunday, September 5, 2010, 8:37 AM


Dear All,
I'm trying to use liver sections in paraffin blocks for IHC, but get no
signal at all, doesn't matter what antibody i'm using.

I'm using section about 3mm thick (the natural thickness of mouse liver) and
about 5-7mm Width and Length.

My fixation protocol is like this:
1) immediately after killing the mouse i'm putting the sections in a
fixation solution that is made from: 10ml formaldehyde 37%, 5ml PBSx20, 85ml
DDW  - pH 7, 24Hr at 4C.
2) ETOH 70%, ETOH 80%, ETOH 96%, ETOH 100% x2 - each for 1Hr at RT
3) Xylen x2 - each for 1Hr at RT.
4) Paraffin x3 - each at 60C for 1Hr.

Before doing IHC:
1) Xylen x2 - each for 10Min at RT
2) ETOH 100%x2, ETOH 96%, ETOH 80%, ETOH 70% - each for 2Min at RT.
3) IHC protocol...

Will Bouin's solution be better ?

Thank you all very much in advance

Itai M
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