RE: [Histonet] oil red in MMA

From:"Patsy Ruegg"

I agree with Gayle on this.  One possible solution might be to try and cut
diamond wire sections without any embedding, but the wire does not cut soft
tissue unless it is completely supported by something hard.  You can
superglue the tissue to a wood block and use water in the wire cooling
troth, but then you would either have to float stain the sections (I have
cut 30 micron thick sections of bone this way) or figure out a way to keep
them on a slide for oil red o staining.  Frozen sectioning is usually the
prefered method for demonstrating fat but cutting frozens on titanium
implants even using the best tungsten carbide knife and the tape transfer
system would be too difficult, I would think. Oh the impossible things we
are asked to do......

-----Original Message-----
[] On Behalf Of Gayle Callis
Sent: Friday, January 07, 2005 10:08 AM
Subject: Re: [Histonet] oil red in MMA

I doubt this would work as the solvents (alcohols, clearants, monomers are
really extracting the lipid, an important factor for making MMA work and
polymerize anyway is to NOT have fat in bone.

The dye would not penetrate the plastic anyway, you would have to remove the
plastic completely and that step would continue to remove the fat from your
tissue plus your sections are very thick.

If you had post fixed your tissue with osmium tetroxide before processing,
you may have had more success in detecting/retaining lipids.

, At 05:42 AM 1/7/2005, you wrote:
>Hi dear histonnetters and happy new year 2005 !!
>does anyone have a protocol of oil red staining in tissu embedded in 
>MMA (mixture of butylmetacrylate and methylmetacrylate)?
>i would like to stain fats in soft tissu on titane. My sections are 
>50Um thikness.
>thank you in advance
>Histonet mailing list

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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