Processing Calcified Bone
|From:||"Jacqueline D. Garfield" |
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,
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