[Histonet] Re: Has anyone tried decalifying bone for frozen sections without fixing first?

[Gayle Callis]

It has been done and is published.   I have not done it.

1) Jonsson R et al.  A demineralization procedure for 
immunohistopathological use:  EDTA treatment preserves lymphoid cell 
surface antigens J Immunological Methods 88:109114,1986.

This group had very good staining after a LONG EDTA decalcification on 
murine hindlegs (14 days) , a rather tedious protocol but their CD markers 
were well stained and tissue was preserved.  After snap freezing EDTA 
decalcified bone, they cut frozen sections, and fixed them with acetone, 
did IHC for T and B lymphocytes, Ia antigens.

2)  Kim L. Kusser and Troy D. Randall
Simultaneous Detection of EGFP and Cell Surface Markers by Fluorescence 
Microscopy in Lymphoid Tissues
J. Histochem. Cytochem., Jan 2003; 51: 5 - 14.

Kusser also cites other publications in her article that may be helpful.






At 09:02 AM 1/28/2005, you wrote:
>Has anyone tried decalifying bone for frozen sections without fixing
>first?
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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)