Chemical method of endpoint decalcification procedure histopathology techniques

There are many methods, which used to check the decalcification of the bones. But the most commonly used method is end-point decalcification. 

It is used to check the calcium concentration in the sample (Bone).

Endpoint Decalcification chemical test Principle

Decalcified tissue (bone) is placed in the decalcifying agents. If tissue is partially calcified then a white precipitate of calcium appears. But if there is a lack of precipitate then the tissue is calcium-free.  

Bone Decalcification checking methods-Histopathology

There are three methods used to assess decalcification.

  1. Physical method
  2. X-ray method
  3. Chemical method

Note: Physical and x-ray methods are cumbersome, therefore the chemical method is the most commonly used method.


Why is it important to determine the endpoint of decalcification?

It is important to check the endpoint decalcification when the sample is partially decalcified. Following consequences will occur in the case of partial decalcification of hard tissue.

  • Poor fixation of that sample
  • Poor staining of that sample.

How do you find the extent of decalcification?

The de-calcification extent of bone or calcified tissue can be checked by the endpoint technique. If their white precipitate appeared after mixing ammonium hydro-oxide OR ammonium oxalate, it means the tissue is incompletely decalcified. We further need a decalcification process.

What is the most recommended test for the completeness of decalcification?

There are three methods for the checking decalcification of various tissues. 

  • Physical
  • X-ray 
  • Chemical

Physical and X-ray methods are time-consuming and expensive, therefore the chemical method is the most recommended test for checking the completeness of decalcification.

Importance of decalcification of hard tissue

When the tissue is partially or incomplete decalcified, it will cause poor staining and poor fixation of that tissue. Therefore if for better results, decalcification is necessary. 

Before further processing tissue, we check that tissue by endpoint technique.

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