[Histonet] cryostat sections of undecalcified tissue implants

[Gayle Callis]

Patsy et al, 

RA Dodds did not embed his bone samples, he coated them with 4% polyvinyl
alcohol (70,000 MW water soluble) then dropped them into a dry ice/Hexane
mixture to snap freeze.  After freezing, he placed bone on dry ice to
evaporate the nasty hexane fumes, and mounted the bones on a metal chuck
with the 4% PVA.  Personally, I do not like this mixture as an embedding
media, OCT works just as well. OCT does have PVA in it, but which one is
proprietary, MW unknown, this is combined with Polyethylene glycol (MW
unknown) and is softer than Dodds method, but with tape transfer, we have
no problems with OCT.   

You can use OCT, and we dilute it 1:1 with water to do the coating for that
method. It works just as well as the 4% PVA.  Most of the time, we embed
bone directly in OCT, which helps surround the sample for support during
sectioning.  

Dodds has a publication in J O Histotechnology  some years back, and John
Tarpley has a wonderful publication last year on sectioning bone with the
Cryojane tape transfer.  The methods are worth reading. 



At 12:45 PM 1/26/2004 -0700, you wrote:
>Liz,
>It depends on how big and crunchy it is.  Dabbs method for cutting fs of
>bone marrow biopsies undecalcified worked pretty well on small samples.  He
>embedded in (I can't think of the name of it off the top of my head) but it
>set up harder than OCT.
>I have done whole rat femurs without decal using the tape transfer system
>and D profile tungsten carbide knives.  I have also done fs on osicles of
>collagen sponges treated with growth factors to stimulate bone and cartilage
>formation.  Some of them at early stages could be cut in the cryostat
>without using the tape and the more developed one's could not.
>Hopes this helps.
>Patsy
>
>-----Original Message-----
>From: histonet-bounces <@t> lists.utsouthwestern.edu
>[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Elizabeth
>Chlipala
>Sent: Monday, January 26, 2004 12:18 PM
>To: histonet <@t> lists.utsouthwestern.edu
>Subject: [Histonet] cryostat sections of undecalcified tissue implants
>
>
>
>Hello all
>
>Is it possible to section undecalcified cryostat sections from implants
>(no plastic) from a rat subcutaneous implant animal model.  The implants
>will have possibly new bone and cartilage, not very dense bone, but some
>bone.  Would I need to use a tape transfer system or could I use a
>tungsten knife and be o.k.  Any suggestions would be appreciated.
>
>Thanks in advance
>
>Liz
>
>Elizabeth A. Chlipala, BS, HTL(ASCP)
>Premier Histology Laboratory, LLC
>P.O. Box 18592
>Boulder, Colorado 80308
>Office: (303) 735-5001
>Fax: (303) 735-3540
>lizchlipala <@t> premierhistology.com
>www.premierhistology.com
>
>Ship to Address:
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>
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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)