Two Separated Protocols with the Most Important Comments for Skeletal Staining in Embryonic and Adulthood Period in Laboratory Animals

introduction: skeletal staining is an important part of anatomical studies which can evaluatemacroscopic disorders in bones and cartilages development. the aim of the present article is toillustrate two distinct protocols by all details in order to stain skeletal system of the lab animalsin all ages of their lives so it becomes easy for future studies in this field.methods: in this paper, staining method was based on sample’s ages. so, all the steps of doubleskeletal staining described in 2 protocols, one for embryos and newborns and another for adults.fixing by ethanol, alcian blue staining, dehydration by ethanol, alizarin red staining, washingby potassium hydroxide and glycerol, storage in pure glycerol were the stages of embryos andnewborns skeletal staining. fixing by neutral formalin buffer, placing the samples in ddh2o,fixing again by ethanol, skinning and eviscerating, alcian blue staining, trypsin digestion,alizarin red staining, potassium hydroxide clearing, koh and glycerol clearing and finallystoring the samples in pure glycerol were the steps of adults staining.results: outcomes of a good and reliable procedure for skeletal staining with alizarin red andalcian blue are the samples whose cartilages and bones are stained blue and red respectively.they are completely transparent so that the skeletal parts can be seen through them.conclusion: it is shown that skeletal staining procedures differ in embryos and adults althoughin the most surveys were tried to use a same protocol in diferent ages of laboratory animals.