Decalcifying Tissue for Histological Processing
What is Decalcification?
Decalcification is usually a routine procedure whose main intention is making calcified tissue compatible with embedding media for cutting a micro slide before staining. In short it is an essential procedure that allows for the removal of calcium deposits especially in bones and tissues.
What Does Decalcification Entail?
Decalcification is very essential for good embedding procedure and in histology it is usually carried out between the fixation and processing steps. The main reason decalcification is used in bones is, it is known to make bones more flexible and easy for pathological examination.
However, some tissues may also contain calcified areas and this is where
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Today a number of procedures have been developed to make decalcification a success. They include immersion in solutions containing mineral acids, organic acids, or EDTA and Electrolysis. These are the most used techniques.
While acids are the most used, strong mineral acids such as nitric and hydrochloric are used with dense cortical bone. These prevents the acids from removing large quantities of calcium at a rapid rate as that can easily damage cellular morphology. Mineral acid decalcifiers are however, not recommended for delicate tissues such as the bone marrow.
For more delicate tissues, organic acids are generally recommended. These include acetic and formic acid. They are usually less aggressive and will act more slowly than mineral acids. This is why formic acid in a 10% concentration has been used as a universal decalcifier.
EDTA is not as harsh as any other acid. Apart from being slow in penetrating body tissues, it is expensive in very large amounts. Finally electrolysis has been tried in experimental procedures but was found to be slow and therefore cannot be used on a daily routine (with huge numbers).
Why Must Fixation Come Before
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