[Histonet] oil red in MMA

[Charles Scouten]

 I am not familiar with diamond wire cutting.  Can you give a reference to learn more about that? 

Another way to cut soft tissue is with a Vibratome.  Superglue the tissue to a pedestal, and cut under water(or media).  50 micron sections is fine, they need to be at least that thick.  Mount on slides, or react further as free floating sections with our hisotdipper apparatus.  Get back to me if this sounds good but you have further questions.


Cordially,
Charles W.  Scouten, Ph.D. 
myNeuroLab.com 
5918 Evergreen Blvd. 
St. Louis, MO 63134 
Ph: 314 522 0300  
FAX  314 522 0377 
cwscouten <@t> myneurolab.com 
http://www.myneurolab.com 


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg
Sent: Friday, January 07, 2005 12:37 PM
To: 'Gayle Callis'; Myri37 <@t> aol.com; Histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] oil red in MMA

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......
Patsy

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Gayle Callis
Sent: Friday, January 07, 2005 10:08 AM
To: Myri37 <@t> aol.com; Histonet <@t> lists.utsouthwestern.edu
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
>Myriam
>
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Gayle Callis
MT,HT,HTL(ASCP)
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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