Alcian Blue Staining of the Mouse Fetal Cartilaginous Skeleton
Adapted from Manipulating the Mouse Embryo, 3rd edition, by Andras Nagy, Marina Gertsenstein, Kristina Vintersten, and Richard Behringer. CSHL Press, Cold Spring Harbor, NY, USA, 2003.INTRODUCTION
The vertebrate skeleton forms by endochondral and intramembranous bone formation. During endochondral bone formation, mesenchyme condensations give rise to cartilages that are eventually replaced by bone. However, there are some permanent cartilages that do not ossify, such as the cartilage of the trachea and articular cartilage of the joints, and intramembranous bone formation occurs directly without a cartilage template. This protocol is ideal for revealing the cartilaginous skeleton of mouse embryos between 12.5 and 16.5 days post-coitum (dpc). The cartilaginous skeleton is stained dark blue by alcian blue, whereas the other tissues of the embryo are made transparent by a clearing solution, benzyl alcohol/benzyl benzoate (BABB).
