Postembedding Decalcification of Mineralized Tissue Sections Preserves the Integrity of Implanted Biomaterials and Minimizes Number of Experimental Animals

Figure 2

Specific osteoclast staining after postembedding decalcification of rat femur. Comparison of undecalcified bone sections with decalcified bone reflects no alteration of bone integrity. (a–c) Preembedding decalcified and paraffin embedded bone samples show TRAP positive cells. (d–f) Undecalcified PMMA section shows the specificity of TRAP staining. (g–i) Postembedding decalcification result is comparable to both decalcified and undecalcified sections. (c, f, and i) Magnified images show the multinucleated osteoclasts on bone surface. Images (a), (d), and (g) were acquired by 5x magnification objective; then individual tiles were stitched together by Leica application suite (LASX) software. Images (b), (e), and (h) were acquired by 40x magnification objective. Images (c), (f), and (i) were acquired by 100x magnification objective.

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