undecalcified bone frozens

From:Gayle Callis <uvsgc@msu.oscs.montana.edu>

You need to access these authors  RA Dodds and CJF van Noorden 

RA Dodds et al, The growing osteophyte: a model system for the study of
human bone development and remodeling in situ  J of
Histotechnology,17(1):37-45, 1994  He has done enzyme histochemistry on
undecalcified bone, plus other technics. He is extensively published,
PUBMED will help. 

How to freeze bone (it has some special considerations, slower snap freeze,
cryoprotection  Aaron JE and Carter J Histochem Cytochem 35:361, 1987 (a

Van Noorden CJF has several publications on bone frozen sections, Cathepsin
B in J Histochem Cytochem 37:617, 1989.  He will cross reference his other

Van Noorden CJF, enzyme histochemical reactions on undecalcified cryostat
sections, Histochemistry 86:127-133, 1986

Hill and Elde J Histochem Cytochem, 38:443, 1990 on improved method for
cryostat sectioning undecalcified bone

You will need tungsten carbide tip insert in steel d profile knives, cold
temperatures, and probably will get the best sections using Instrumedics
CryoJane frozen section tape transfer device, www.instrumedics.com.  It
also depends on the cryostat you have, Hacker has special set up Dodds
used, but that may not be your situation. 

We do all bone undecalcifed frozens with Instrumedics tape transfer, too
difficult without this instrument.  Gave up using antiroll device, picking
up sections, etc long ago

Go to Histonet Archives, there is much discussion on bone frozen sections -
the difficulty, how to snap freeze the bone, hints on sectioning, etc, etc.
 Go to 


You will also need a great deal of patience to cryosection bone!

Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303

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