In Situ Hybridization of Bone and Cartilage

Abstract

In situ hybridization is a technique that combines histology and molecular biology. In situ hybridization provides cell-type specific expression of genes along with their histological characteristics. This chapter is intended for beginners in either molecular biology or histology who want to begin in situ hybridization histochemistry. The chapter examines equipment and reagents that are specific for in situ hybridization; preparation of specimens (dissection, fixation, decalcification, embedding and sectioning); preparation of probes (design of probe, labeling, storage); hybridization (pretreatment of sections, hybridization, washing); and detection of the hybridized probe. In addition to the introduction of a general method used in the authors’ laboratory, several optional or alternative protocols are described (see Table 1).