Hi, I'm doing a project on the embryonic development of bearded dragons, and am currently decalcifying a few specimens in preparation for serial sectioning. So far I've been having calcium detected in surprising places - for instance a tail snip from a 12 day old embryo was found to have calcium, even though these apical vertebrae shouldn't form until much later one.
Does anyone have any experience with this? Namely, could the formalin in which they were preserved for quite some time have caused calcium leeching?
Thanks!
Dear Sarah,
1. Soluble calcium salts move during the formalin fixation following the concentration gradient.
2. Insoluble calcium salts may be dissolved in the acid components present in the fixatives, unbuffered formalin fixative has low pH, it contains acids such as formic acid.
3. The decalcification solves most insoluble calcium salts and depending on the decalcifying solution composition, procedure duration, and washing, some calcium compounds may be redistributed in the object.
Best regards
Pavel
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