[Histonet] Re: TRAP staining on formic acid decalcified bone reference

[Gayle Callis]

The reference within a reference from Ray is A Chimeric Form of
Osteoprotegerin Inhibits Hypercalcemia and Bone Resorption Induced by IL-1β,
TNF-α, PTH, PTHrP, and 1,25(OH)2D3 .   Sean Morony et al .  J Bone Mineral
Res V 14, pp 1478-1485.  

 

However, the formic acid decalcification method is not described in detail
and merely says "formic acid" but whether this is buffered formic acid or
just dilute formic acid in water only is not stated.     Ray might elaborate
on what specific formic acid recipe he used as many in research don't always
use buffered formic acid decalcifiying solutions.  

 

I would assume Morony et all used a buffered formic acid with either sodium
formate or sodium citrate and controlled so as to not overexpose TRAP to
acids longer than necessary.   One publication,  i.e.,  Eggert and Germain.
Stable Acid Phosphatase I. Demonstration and Distribution.  Histochem 66, pp
301-317, 1980) discussed in detail the  rapid demineralization in acidic
buffers i.e. buffered formic acid for staining of stable forms of acid
phosphatase.  

 

I have both of these publications on file and will forward privately.   

 

I would err on the side of using a buffered formic acid with either sodium
formate or sodium citrate for doing this and use decalcification endpoint
testing to avoid over exposure to acid i.e. over decalcification.    

 

Take care

 

Gayle M. Callis 

HTL/HT/MT(ASCP)