Re: [Histonet] Supplies needed/protocols for

From:Gayle Callis

Methyl methacrylate is superior - first and foremost - you need fume hood
and careful handling to avoid toxic fumes. 

YOu will need a POWERFUL, hopefully automated microtome, Leica 2155 or
2165, Tungsten carbide knives (more than one from Delaware Diamond Knives,
d profile in a universal type knife holder that can be used for c profile
steel knives also. A vacuum system or pump with vacuum desiccators (must
work in a fume hood vented to outside is best, fumes are very toxic to
Central nervous system), refrigerator, final block should be shaped with a
saw or grinding to fit into block holder, MMA blocks are very hard and can
be cut to fit into a screw down block holder. An oven to cure to final
block )after polymerization. 

You cannot use conventional molds very easily, as evaporation is too great
and this creates problems with plastic. You need to place samples inside a
"mold", unusally invented by you! Round polypropylene containers to fit
legs into with a screw on lid. Containers that hold samples in formalin can
be used as they have lids that with screw or snap on,  plus you may need to
do prepolymerized layers - a nice way to have flat bottom blocks.    

A waterbath for polymerization for even heating at 37C for polymerization,
doing this in an oven will cause fumes and bubble formation. 

I suggest you start reading about methyl methacrylate, Journal of
Histotechnology has had some superb publications over the last 15 years, by
Sterchi, Mayton(Sanderson), Chappard, Hand, Jenkins, and others.  

Lots of patience is needed to work with MMA - there are some workshops this
year at National Society of Histotechnology, Toronto Canada, Ruth
Yaskovitch and another lady are presenting a very good one on just what you
need.  It would be a good investment to take a workshop like this, you get
so SEE and HEAR about MMA methods which helps you avoid problems. 

Good luck


 At 09:51 AM 6/7/2004 -0400, you wrote:
>Please tell me what supplies I need to begin undecalcified bone
processing. I have addressed the aquisition of knives, knife holder, and
block holder.
>    What I need now are;
>    1. Which plastic is best for bone?
>    2. What kind of embedding molds and what sizes do they come in?
>        (We are going to do rat legs)
>    3. What kind of instruments will I need?
>I'm new at this and feeling my way, please HELP!
>                        Thanks
>                             Thelma
>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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