Re: [Histonet] immunostaining of VEGF and CD31 in mouse decalcified bone tissue

From:Gayle Callis

Publication on bone using EDTA on fresh bone before snap freezing and
fixation of a frozen section.   

Mori S et al.  A new decalcifying technique for immunohistochenical studies
of calcified tissue especially applicable to cell surface marker
demonstration, J Histochem Cytochem 36:111-114, 1988. 
Abstract: We have developed a new decalcifying technique for use in light
and electron microscopy studies utilizing immunohistochemical staining of
calcified tissues. Specimens containing calcified tissues can be adequately
decalcified at a pH of 7.1-7.4 and temperature of -5 degrees C, without
freezing, by use of a solution containing EDTA, sodium hydroxide, and
glycerol. In this study, Leu-2a, Leu-3a, Leu-4, Leu-7, Leu-12, Leu-14,
Leu-M1, Leu-M2, Leu-M3, and HLA-DR-positive cells in destructive lesions of
bone tissues from patients with rheumatoid arthritis and osteomyelitis were
successfully detected immunohistochemically. Furthermore, the presence of
HLA-DR antigen on the surface of the infiltrating cells in the same lesions
could be demonstrated using the immunoelectron microscopic technique. The
results reported here have not previously been obtainable using
conventional decalcifying techniques. 

Be sure to search in this journal, there have been some excellent, more
recent publications on decalcificatin of bone to do IHC on CD markers, etc.  

Gayle Callis
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)

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