Skeletal Examination by Alizarin Staining

Skeletal Examination by Alizarin Staining 213 Pair of arches of I st sacral vertebra... 

Pregnant females were sacrificed by carbon dioxide anesthesia on day 21 of gestation the uterine horns were exteriorized through a midline incision in the abdominal wall, and the number and position of live, dead, and resorbed fetuses were noted. After being weighed and sexed, the fetuses were examined for external anomalies the crown-rump length was measured with a vernier caliper. Half of each litter was preserved in Bouin s solution and later examined for soft tissue anomalies (16) the other half was preserved in alcohol, cleared and stained with Alizarin red-S, and examined for skeletal abnormalities (2). 

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