There are many hard calcium tissue in pathological experiments. The density between calcium and paraffin in tissue are different, and tissue containing calcium can’t be sliced directly. Bone tissue contains the most calcium and calcification may occur in other tissues forming calcified areas in the tissue, it needs to decalcify. Decalcification is an important step in making bone slices, especially the bone tissue to immunohistochemical stained.
EDTA can combine with calcium in the outer layer of hydroxyapatite crystal to form soluble non-ionic compounds, and promote the outward transfer of bound calcium in the inner layer of the crystal. With the continuity, the hydroxyapatite crystal is gradually melted, and it can play a chelating role when the pH is neutral. This method is characterized by long decalcification time, less damage to bone tissue, better preservation of enzyme activity (alkaline phosphatase) and cell antigenicity. The slices can be used for histochemical and immunohistochemical analysis.
Decalcification rate will be slower but preservation of cellular morphology is excellent and viability of staining for enzymes, immunohistochemistry antigenicity and electron microscopy is maintained.
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