Re: Cryosectioning embryonic mouse spinal cord

From:Gayle Callis

Your embryos (you did not give age in days) will require no
decalcification.  You can embed the whole verterbral column for midsaggital
(long axis) sectioning OR dissect out the column and place various
vertebral sections on end for transverse i.e. cross sections of cord.   

Just snap freeze appropriately, and if the cord is friable during
sectioning at -20C, warm up cryostat to -16C and cut.  

Use a good, sharp disposable blade for optimal sections, and let us know
what kind of results you get, embryos are always more difficult to work with. 

If you prefix in paraformaldehyde or NBF, be sure to cryoprotect in 20 to
30% sucrose overnight before snap freezing.  Snap freeze with
isopentane/dry ice slurry or liquid nitrogen cooled isopentane or a plastic
petri dish floating on liquid nitrogen, set mold in dish (do not put liq N2
INSIDE the dish), let it freeze for freezing artifact free tissues.  Tissue
Tek cryomolds (not any others please where the plastic is too thick) are
superb for flat, well oriented tissues. 

Have fun

 At 10:06 AM 7/2/2003 -0600, you wrote:
>Dear netters-
>I need to cryosection embryonic mouse Spinal cords.  I would like to embed
>them in TissueTek OTC.  Any suggestions, tips, warnings?
>Thanks in advance
>David Walker
>UM Missoula
>Missoula MT
Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
S. 19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367 (lab with voice mail)
406 994-4303 (FAX)


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