[Histonet] histogel, mouse bile duct embed/freeze

[Gayle Callis]

You can use OCT and a Tissue Tek plastic cryomold.  

We freeze lengths of mouse spinal cord by laying them flat on the bottom of a 
Tissue Tek cryomold (whatever size works best). You can coat the tissue
with bit of OCT, lay tissue in bottom of mold, add more OCT and snap freeze
but you must hold the mold flat so the bottom remains uncurved, perfectly
flat.  After freezing, one can pop the block out, orient the frozen tissue
block ON END for  cross sections, and freeze onto metal chuck in that
orientation.  More OCT can be added at that time to build up block face, a
gradual process to not thaw block. It is more like a double embedding.  

Our flattest blocks are made by sitting the cryomold with OCT embedded
tissue in the bottom of a plastic petri dish that floats on liquid
nitrogen. Do not allow Liq N2 to go INSIDE the dish.  The dish can sit on a
platform immersed in liquid N2 so the dish cannot tip over or allow Liq N2
into the dish. 

WE get perfect cross sections of spinal cord with this type of embedding
and no freezing artifact. 



 At 01:36 PM 10/2/2003 -0600, you wrote:
>Has anyone ever tried using something like histogel to orient tissue for 
>frozen sectioning?  I am wondering about the cutting and freezing quality of 
>samples held in histogel til firm and then quick frozen in oct?  I need to 
>get cross sections of mouse bile ducts.  I do this very successfully using 
>histogel and paraffin processing, but I need to get frozen sections of the 
>same for IHC T subset studies.
>Please advise.
>Best regards,
>Patsy
>
>
>
>Patsy Ruegg, HT(ASCP)QIHC
>IHCtech, LLC
>Fitzsimmons BioScience Park
>12635 Montview Blvd. Suite 216
>Aurora, CO 80010
>P-720-859-4060
>F-720-859-4110
>HP-303-644-4538
>HF-303-644-3377
>hm email pruegg <@t> msn.com
>wk email pruegg <@t> colobio.com
>
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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)