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

[Jennifer MacDonald]

We use pre-fixed tissue for oil red o.  We rinse the section briefly to 
remove the formalin from the outside of the tissue.  We prepare our tissue 
in OCT or the equivalent.  We cut the frozen sections and mount them on 
charged slides.  We let dry the dry a minimum of 10 minute, rinse them in 
water to remove the OCT and then proceed with the stain.




kyoung LEE <east78 <@t> yahoo.co.kr> 
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
04/14/2011 08:04 AM

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[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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