Dear Histonetters,

I'm in search of processing protocols for calcified bone.   The samples for
this particular study are calcified sheep bone with tendon attached.  The
average sample size is 1cm X 1cm X 0.3mm.  Also, these samples have been
labeled with tetracycline dye.  

We are currently trying three different fixatives (NBF, Alcohol Formalin,
and 70% ETOH).   We plan on dehydrating in 70%, 95%, and 100% ETOH, for
about 4 days each; followed by xylene for 24hours.  

Then we have a dilemma?  What embedding media should we infiltrate and embed
with?  PMMA?  MMA?  or  JB-4?   And for how long?  I am having difficulty
finding protocols, especially for the PMMA method.  

Once embedded, we hope to be able to obtain 5 µ sections on a Leica RM 2155
microtome with a D-profile Tungsten Carbide Knife.  

Are we dreaming?  Or does this sound somewhat reasonable?  Your thoughts,
advise, prayers, and protocols are greatly appreciate.  

Thanks in advance,
Jackie Garfield