Rapid Method of Preparation of Large Specimens of Undecalcified Bone and Teeth for Sectioning at a 4-Micron Thickness

¹ Radioisotope Service, Veterans Administration Center, Wood, Wisconsin

An improved, rapid method for imbedding teeth or large specimens of undecalcified bone for sectioning at a 4-µ thickness has been developed. The specimen is dehydrated with alcohol and ether. Then it is treated with a 1:1 mixture of monomers, ethyl, and n-butyl methacrylate, catalyzed with 2,4-dichlorobenzoyl peroxide, and again with ether. After the ether is removed under vacuum, the specimen is impregnated with the partial polymer of the above monomers under vacuum in a vacuum dessicator. A special pressure chamber has been developed that facilitates rapid, safe curing of the specimen under pressure and heat. Polymerization is carried out in the chamber for 72 hours at 60°- [see figure in the PDF file] 70° C. followed by 4 hours at 90° C. Sectioning is done on a Jung Model K Microtome at 4-µ thicknesses for both microscopic and autoradiographic study. Excellent structure is evident and can be easily discerned from non-calcified matter.