[Histonet] [Help] oil red O stain in fattty change liver

[Setlak, Lisa]

We do this stain on frozen sections in our lab with no formalin fixation. We stain in oil red o from American Mastertech for 30 minutes, rinse in water, counterstain in hematoxylin, coverslip with aqueous mounting media.
Lisa

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of kyoung LEE
Sent: Thursday, April 14, 2011 9:07 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] [Help] oil red O stain in fattty change liver

Dear sirs.

I don't know whether I send this question to you.

For several weeks, I'm setting oil red O stain in fattty change liver (mouse).

Could you review my protocol?

1. Murine tissue,livers, is fixed with 10% NBF(10% neutral-buffered formalin).
2. Make frozen block 
3. Blocks were cut 5um in crystat and dried for 1~2H at RT. 
4. store at -20 
5. let it dry for 30mins at RT
6. immerse it in cold 10% NBF for 10mins
7. staining

After staining, my section were fallen off and seperated.

I search many protocols. Some suggest additional fixation is necessary as soon 
as cutting.
Others suggest prefixed tissue is not necessary additional fixation.

Plz send me your comments.
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