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The hematoxylin/eosin (H&E) staining and Masson’s trichrome staining

RZ Ruotian Zhang
MY Min Yang
YL Yang Li
HL Hedong Liu
MR Maoxian Ren
ZT Zhou-Shan Tao
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Samples were immersed in fresh 10% neutral formalin for 72 h at ambient temperature. Femur specimens were decalcified in 15% neutral EDTA buffer for 3 months. After the decalcification was completed, the decalcified specimens were sequentially placed in different concentrations of ethanol for a gradient dehydration. The bones were then embedded in methyl methacrylate and cut into 5-mm sections for hematoxylin and eosin (H&E) and Masson’s staining [36]. All the samples were prepared and examined in a blind manner to reduce experimenter bias. New bone formation area (%) = new bone formation area/bone defect area × 100%.

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