[Histonet] Canine bone sectioning trouble

[Schneider,Lynda S]

Hello out there...

We are having trouble sectioning canine bone.  The samples are bone marrow cubes approximately  2cm thick and 1in wide.  They were fixed for 15 hours in 10% NBF and decaled for 30 - 45 mins in RDO rapid decalcifier.  They were faced in and then surface decaled again for about 30 mins.  When sectioned, much of the marrow was missing and the bone was torn and shredded.  We thought that maybe the samples had not been fixed sufficiently so refixed overnight in 10% NBF again.  The samples were then reprocessed as they had been originally.  The processing schedule used was: 70% EtOH 30mins, 80% EtOH 30mins, 95% EtOH x 2, 30mins each, 100% EtOH x 2, 30 mins each, xylene x 2, 45 mins each, paraffin x 4, 40 mins each. Again they were faced in and decaled for another 30 mins or so.  This time as soon as the sections are placed on the water bath (38 degrees) they explode and/or come apart so severely sections can almost not be picked up.  If sections are even obtainable they are of horrible quality.  Does anyone have any suggestions? Thank you so much in advance!

Lynda