Staining and histochemistry of undecalcified bone embedded in a water-miscible plastic.
Undecalcified bone fixed in a variety of fixatives and embedded in a new formulation of 2-hydroxypropyl methacrylate at 4 c has been sectioned at 1 to 5 microns. The embedding mixture contains 2-butoxyethanol as plasticizer and triethyleneglycol dimethacrylate as cross-linker. The accelerator was benzoyl peroxide and the catalyst was N,N-dimethylaniline. With proper embedding and care in sectioning it is possible to obtain sections with relatively little bone compression, excellent preservation of cellular elements, and a minimum of wrinkling. A wide variety of stains have been used for these sections and those reported here are Gill's hematoxylin-eosin, Nocht's azure-eosin, Feulgen, Hoechst 33258 (bisbenzimid H 33258), methyl green-pyronin, PAS, alizarin red, and von Kossa silver stain. There was excellent preservation of acid and alkaline phosphatase activities. A new method of prestaining immunofluorescent labeling was also applied to bone and examples of staining with anticollagen I and antifibronectin are presented.
D000818:Animals; D001842:Bone and Bones / Q000032:analysis* / Q000033:anatomy & histology; D002642:Chick Embryo; D002645:Chickens; D003094:Collagen / Q000032:analysis; D005353:Fibronectins / Q000032:analysis; D006651:Histocytochemistry; D006652:Histological Techniques; D008867:Microtomy; D010744:Phosphoric Monoester Hydrolases / Q000032:analysis; D013194:Staining and Labeling / Q000379:methods*
D005353:Fibronectins; D003094:Collagen; D010744:Phosphoric Monoester Hydrolases