“…Injections can be made or operations performed on a specific portion of the embryo, and resultant changes in heart rate, behavior, regional growth, or other parameters can be observed continuously without having to sacrifice the embryo (B. E. Dunn, Fitzharris, et al, 1981). Additionally, agents which affect angiogenesis (Ligresti et al, 2012) or vasoreactivity (L. K. Dunn et al, 2005; Harland et al, 2012) can be placed onto the chorioallantoic membrane (CAM), and the effects can be followed for multiple days without having to sacrifice either host or graft tissue. Since in ovo approximately 75% of the calcium in the newly hatched chick is derived from the eggshell (B. E. Dunn & Boone, 1977; Ono & Wakasugi, 1984), shell‐less culture provides a unique opportunity to study the effects of calcium metabolism in the developing embryo.…”