Large-scale double-staining of rat fetal skeletons using Alizarin Red S and alcian blue.
A critical component in the conduct of a prenatal developmental toxicity study is the evaluation of fetal skeletal development. As the developing rodent fetus is typically evaluated at gestation day 20, at a time when ossification of the skeleton is incomplete, a thorough assessment of skeletal development would include both ossified and cartilaginous structures. Current methods to double-stain the fetal skeleton using Alizarin Red S and Alcian Blue are typically described for small sample sizes or using time allotments for each processing step that are unsuitable for industry. In an industrial setting, there is a need for an effective means to double-stain fetal skeletons on a large scale (i.e., hundreds of fetuses simultaneously). This article describes a method used in our laboratory to stain both fetal bone and cartilage using solutions and procedures on an industrial scale.
Young
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Bayer Corporation, Agriculture Division, Toxicology Department, Stilwell, Kansas 66085, USA. angela.young.b@bayer.com
Phipps
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Astroff
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D000423:Alcian Blue; D000818:Animals; D000880:Anthraquinones; D001842:Bone and Bones / Q000166:cytology / Q000196:embryology*; D004396:Coloring Agents; D005314:Embryonic and Fetal Development; D005260:Female; D005865:Gestational Age; D008297:Male; D010012:Osteogenesis; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley
D000880:Anthraquinones; D004396:Coloring Agents; C004468:Alizarin Red S; D000423:Alcian Blue