An Improved Hard Tissue Slicing Technique：With Potential in the Fast Clinicopathological Diagnosis of Bone Diseases

Abstract

The specimens of bone are often embedded in paraffin for pathological diagnosis in the clinic. To obtain sections with complete biological signals, bone tissue needs to be decalcified before embedding. However, decalcification takes a long time, which could easily lead to protein denaturation, enzyme activity reduction, and loss of immune reactivity. The plastic-embedded sections of non-decalcified bone revealed a better tissue morphology and a more robust positive immunoreaction than paraffin-embedded sections. However, the traditional hard tissue slicing method is complicated and difficult to promote. In this study, osteosarcoma, fibrous dysplasia, and femoral head necrosis samples were treated with an improved resin-embedded sectioning method compared with the same sections embedded in paraffin. The production process of the improved method is simple. The immunohistochemical markers, such as CD34, Ki67, and SATB2, exhibited more accurate intracellular localization after anti-stripping treatment. Furthermore, MDM2 detected by fluorescence in situ hybridization (FISH) and GNAS of mutational analysis, treated with the improved resin-embedded method, met the pathological diagnosis requirements. The fixation, dehydration, embedding, sectioning, and staining of rigid tissues were all standardized. The improved hard tissue slicing method can be applied in the laboratory and has the potential for clinicopathological diagnosis.

Note:

Funding Information: This work was supported by the National Natural Science Foundation of China (grant no. 81802247, 81702181, 81972500) and the Projects of International Cooperation and Exchanges of National Natural Science Foundation of China (grant no. 81820108020).

Declaration of Interests: The authors declare no conflicts of interest.

Ethics Approval Statement: The study protocol complied with the Declaration of Helsinki and was approved by our hospital’s Human Research Ethics committee (Shanghai Jiao Tong University School of Medicine Affiliated Sixth People’s Hospital).

Keywords: hard tissue slicing techniqueplastic-embeddedbone tumorfemoral head necrosis