Visualization of Different Tissues Involved in Endochondral Ossification With Alcian Blue Hematoxylin and Orange G/Eosin Counterstain

Jessica R. Nowalk, Lisa M. Flick
2008 Journal of Histotechnology  
The histologic evaluation of endochondral ossification is critical to the study of fracture healing, developmental biology, and comparative histology. A modification of Sayers' alcian blue hematoxylin staining technique is described in which a different counterstain is applied. Paraffin sections are stained in alcian blue hematoxylin for 30 min, and then placed in orange G-eosin counterstain for 1 min, 30 s. This method can differentiate cartilage, mature bone, and immature bone found in
more » ... stages of endochondral ossification and fracture callus. Increasing the hematoxylin content to 0.5% produced a more striking contrast between the bone and cartilaginous aspects of the healing fracture callus. Inclusion of orange G (instead of acid fuchsin) in the counterstain provides better demarcation of mature and immature bone. Alcian blue hematoxylin/orange G-eosin consistently stains cartilage blue, mature bone orange, and immature bone mauve and is particularly suited to the study of endochondral ossification, fracture healing, and bone remodeling. (The J Histotechnol 31:
doi:10.1179/his.2008.31.1.19 fatcat:axtgg3co4bhwlk7xpgvsjvbltq