The purpose of this study was to investigate the in situ hybridization technique of paraffin-embedded skeletal tissues by non-radioactive labelling oligonucleotide, Five six-week-old male SD rats were selected and the proximal epiphyseal growth plates of bilateral tibiae were removed to be decalcified in 0.5 mol/L EDTA. The tissues were embedded by paraffin and the sections were treated and hybridized with RNase-free reagents. Control groups were set to testified the sensitivity and the specificity of the experiment. The results showed that this method preserved the integrate structures and effectively detected the mRNA of the tissues. These indicated that it was an efficient and practical approach to the biomolecular research of skeletal tissue.