Alterations to the en bloc basic fuchsin staining protocol for the demonstration of microdamage produced in vivo.

En bloc staining with basic fuchsin has become the method of choice to demonstrate bone microdamage produced in vivo. Several alterations have recently been made to the protocol. This technical note presents the new protocols, which include staining through a graded series of alcohols under vacuum, and eliminating the original evaporation stage in the staining protocol. Reasons for variations in staining quality can be related to inadequate dehydration, failure to stain under vacuum, the source of the basic fuchsin, or the solubility of basic fuchsin in water. The most common reasons for over- or understanding are the time left in stain, and the density of the bone. Limitations to the technique include the fact that probably not all cracks are stained by the technique, and the technique is not useful for studies that involve bone with highly variable densities in a single section.