Re Oil red staining

From:RichardWHorobin@aol.com


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Jaclynn Lett
wrote:

> Has anyone used Oil Red O to stain lipids in tissues embedded in plastics 
> (Epon or Epon-Araldite)?  If so, has this been done by staining en bloc or 
> by staining the sections.  Sections would range from 1-4 microns in 
> thickness.  

A couple of people so far have commented on how hard this would be, and our 
experience agrees with theirs.
HOWEVER Jaclynn also said:


> We would also consider tissues embedded in glycol methacrylate.  We'd like 
> to avoid frozen sections because we'd prefer the higher level of detail 
> possible with plastic. 

In THAT case things look better! Would you settle for Sudan black, rather 
than Oil red staining of the lipid? If 'yes' then there is a method - which 
does indeed show even tiny droplets of lipid very clearly. This was worked 
out by the one-time king of GMA staining Peter Gerrits, and can be found in J 
Neurosci Methods, as follows:

      Gerrits PO, Brekelmans-Bartels M, Mast L, 's-GrAavenmade EJ, Horobin RW 
and Holstege G. (1992).. 
      Staining myelin and myelin-like degradation products in the spinal 
cords of chronic experimental 
      allergic encephalomyelitis (Cr-EAE) rats using Sudan Black B staining 
of glycol methacrylate-embedded 
      material. 
      J. Neuroscience Methods. 45, 99-105

Bye - Richard Horobin

Institute of Biomedical & Life Sciences, University of Glasgow
T direct 01796-474 480 --- E  RichardWHorobin@aol.com
"What should we expect? Everything."

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<HTML><FONT FACE=arial,helvetica><FONT  SIZE=2>Jaclynn Lett
<BR>wrote:
<BR>
<BR><BLOCKQUOTE TYPE=CITE style="BORDER-LEFT: #0000ff 2px solid; MARGIN-LEFT: 5px; MARGIN-RIGHT: 0px; PADDING-LEFT: 5px">Has anyone used Oil Red O to stain lipids in tissues embedded in plastics 
<BR>(Epon or Epon-Araldite)?  If so, has this been done by staining en bloc or 
<BR>by staining the sections.  Sections would range from 1-4 microns in 
<BR>thickness.  </BLOCKQUOTE>
<BR>
<BR>A couple of people so far have commented on how hard this would be, and our 
<BR>experience agrees with theirs.
<BR>HOWEVER Jaclynn also said:
<BR>
<BR>
<BR><BLOCKQUOTE TYPE=CITE style="BORDER-LEFT: #0000ff 2px solid; MARGIN-LEFT: 5px; MARGIN-RIGHT: 0px; PADDING-LEFT: 5px">We would also consider tissues embedded in glycol methacrylate.  We'd like 
<BR>to avoid frozen sections because we'd prefer the higher level of detail 
<BR>possible with plastic. </FONT><FONT  COLOR="#000000" SIZE=3 FAMILY="SANSSERIF" FACE="Arial" LANG="0"></BLOCKQUOTE>
<BR></FONT><FONT  COLOR="#000000" SIZE=2 FAMILY="SANSSERIF" FACE="Arial" LANG="0">
<BR>In THAT case things look better! Would you settle for Sudan black, rather 
<BR>than Oil red staining of the lipid? If 'yes' then there is a method - which 
<BR>does indeed show even tiny droplets of lipid very clearly. This was worked 
<BR>out by the one-time king of GMA staining Peter Gerrits, and can be found in J 
<BR>Neurosci Methods, as follows:
<BR>
<BR>      Gerrits PO, Brekelmans-Bartels M, Mast L, 's-GrAavenmade EJ, Horobin RW 
<BR>and Holstege G. (1992).. 
<BR>      Staining myelin and myelin-like degradation products in the spinal 
<BR>cords of chronic experimental 
<BR>      allergic encephalomyelitis (Cr-EAE) rats using Sudan Black B staining 
<BR>of glycol methacrylate-embedded 
<BR>      material. 
<BR>      J. Neuroscience Methods. 45, 99-105
<BR>
<BR>Bye - Richard Horobin
<BR>
<BR>Institute of Biomedical & Life Sciences, University of Glasgow
<BR><B>T direct 01796-474 480 --- E  RichardWHorobin@aol.com</B>
<BR><I>"What should we expect? Everything."</I></FONT></HTML>

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