Comparative study on neutralization methods of decalcified bone specimen: between rinsing with running tap water and immersing in saturated lithium carbonate solution

Abstract

Background : Neutralization of decalcified bone specimens is required for removing excess acid, resulting in better hematoxylin and eosin (H&E) staining.

objective : The aim of this study is to compare between the conventional method and chemical neutralization method using saturated lithium carbonate solution.

Methods : Two groups of decalcified bone specimens were treated with two different neutralization methods: i) rinsing with running tap water; and ii) immersing into a saturated lithium carbonate solution. Both groups were subsequently subjected to H&E. The quality of staining of the two groups was evaluated by two board certified pathologists. 

Results : Comparing to the conventional method, H&E-stained tissues from the lithium carbonate method tended to yield better results in terms of: i) staining quality with strong contrast and ease of visualization; ii) clarity of cellular details and stained portions of the cells; and iii) clarity of nuclear morphology.  

Conclusion : Neutralization of decalcified bone specimens by immersing into saturated lithium carbonate solution resulted in better H&E staining than the existing method. Moreover, employing lithium carbonate to neutralize bone tissues can lower water bills and assist with water conservation in our area. 

Keywords : Neutralization, post-decalcification, saturated lithium carbonate solution, Bone tissues