Staining and histochemistry of undecalcified bone embedded in a water-miscible plastic

Stain Technol. 1980 Sep;55(5):313-21. doi: 10.3109/10520298009067260.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone and Bones / analysis*
  • Bone and Bones / anatomy & histology
  • Chick Embryo
  • Chickens
  • Collagen / analysis
  • Fibronectins / analysis
  • Histocytochemistry
  • Histological Techniques*
  • Microtomy
  • Phosphoric Monoester Hydrolases / analysis
  • Staining and Labeling / methods*

Substances

  • Fibronectins
  • Collagen
  • Phosphoric Monoester Hydrolases