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Decalcification using EDTA

EDTA Solution:

0.5M EDTA pH 8.0 (GIBCO) 30ml

ddH₂O 70ml

6N HCl 1.2ml

pH 6.4-6.8

1. Wash fixed tissure with PBS 20min X3.
2. Wash tissure with ddH₂O 20minX3.
3. Change tissure to EDTA solution with agitation at RT for 2-3 days depend on tissure size.
4. Change to fresh EDTA solution if need further decalcification.
5. Rinse with ddH₂O several times.
6. Wash with ddH₂O 20minX3
7. Dehydration and embedding ( See histology protocol)

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