Protocol for decalcification.

[Clas Johansson]

I have for some time labelled mitotic cells in the CNS of rodents 
without any problems. Now I would like to do the same thing with 
adult Zebrafish. I have administrated BrdU to the fish and fixed 
them with 4% PFA. Put them in 30% sucrose before cutting on a 
cryostat. My problem is: since the spinal cord is so small it was 
difficult to get it out (so I decided to cut the fish in one piece) 
however, this was very difficult because the bone around the spinal 
cord tear the spinal cord apart. So, does anyone have a protocol for 
decalcification or anything else that can help me out?  

Best regards Clas
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Clas B. Johansson, DDS, Leg. Tandl=E4kare
Karolinska Institute
Medical Nobel Institute
Department of Cell and
Molecular Biology
von Eulers v=E4g 3
SE-171 77  STOCKHOLM
SWEDEN
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Phone +46 8 728 74 96, 75 13
Fax: +46-8-348135
Email: clas.johansson at cmb.ki.se