[Histonet] Oil Red O despair

[Andrea Grantham]

I get many projects here that require frozen liver sections and Oil 
Red O staining. Some years ago I was having problems with the 
staining and someone on histonet suggested the PolyScientific R & D 
Oil Red O Stain Kit. I got the kit and have been using it ever since. 
The stain works great - usually I have livers loaded with lipid - and 
the slides come out clean. The kit uses proplyene glycol and glycerin 
jelly to mount the coverslips. I do use one part of the ORO protocol 
from Freida Carson's book - I fix the slides in 37-40% (page 152, 
second edition).

Andi Grantham




At 04:49 PM 11/17/2008, Amos Brooks wrote:
>Michele,
>    You could try 0.5% Oil Red O in 60% triethylphosphate. That works really
>well here. I've also used a formulation I found in Humanson, the same amount
>of Oil Red O in 99% ethanol and it worked fairly well. Triethylphosphate is
>much cleaner than the ethanol. (There was a precipitate on the slide.) I
>can't avouch for isopropyl as I used ethanol, but it is good to know there's
>an alternative in case we run out of triethylphosphate.
>
>Amos Brooks
>
>Message: 10
>Date: Mon, 17 Nov 2008 11:56:45 -0600
>From: "Michele Wich" <mwich <@t> 7thwavelabs.com>
>Subject: [Histonet] Oil Red O despair
>To: <histonet <@t> lists.utsouthwestern.edu>
>Message-ID:
>        <62A8156F8071C8439080D626DF8C33A602E554 <@t> wave-mail.7thwave.local>
>Content-Type: text/plain;       charset="us-ascii"
>
>I'm wondering what are the pros (or cons) of using the propylene glycol
>version of Oil Red O over the isopropanol method. Is one more suited to
>a specific application?
>
>I'm trying to stain a frozen section of liver, which one would suspect
>would be loaded with fat, and have thus far been unsuccessful using the
>propylene glycol Oil Red O.
>
>Is there something obvious that I'm missing here? I'm new to
>cryosectioning...perhaps I did something wrong in the cryostat. I fixed
>my sections in NBF before staining.
>
>Please help. I'm feeling like a pathetic excuse for a histotech. Any
>advice is greatly appreciated.
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.....................................................................
: Andrea Grantham, HT(ASCP)     Dept. of Cell Biology & Anatomy     :
: Sr. Research Specialist       University of Arizona               :
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