Manual for the Preparation and Staining of Embedded Natural Dry Bone Tissue Sections for Microscopy

Abstract

In the last decade, the use of light microscopy has been firmly established for the investigation of exhumated human bone tissue. As a rule, these remains cannot be decalcified; thus, they are most commonly prepared for microscopic analysis as ground thin sections. These ground sections are of great value in diagnosing disease, in estimating age or in assessing taphonomic alteration. As bone is sometimes fragile and can be damaged by the grinding process, the specimen is occasionally supported by an embedding medium. In contrast to the vast amount of research conducted on embedded and unembedded unstained bone material, the use of histological stains on undecalcified dry bone tissue has been long neglected. In this article, a new method for embedding, sawing and grinding dry bone tissue is presented. The produced sections are subsequently stained with haematoxylin. The results show that even ground sections of fragile bone can be made in a quick and easy manner. Staining these sections enhances the envisioning of microarchitecture and taphonomical processes. In addition, the sections stay open for inspection under polarized light. The results were consistent throughout the used bone material. To keep the method as accessible and comprehensive as possible, a step-wise manual is provided. An additional troubleshooting paragraph discusses the most often encountered problems and provides solutions.

Healed antler fracture from a giant deer (Megaloceros giganteus) from the Pleistocene in Poland

“…Based on radiographic images, three antler foci were selected for histological analysis: (a) a radiographically normal site ( (2003), with the modifications of de Boer et al (2013). Unstained slides (a crosssection of the antler, 20-30 μm in thickness) were examined histologically using light microscopy.…”

Paleohistology and the study of human remains: past, present and future approaches

“…Beauchesne and Saunders, 2006;Martiniaková et al, 2006). Recently, a new method for embedding, sawing, grinding, and staining was proposed by De Boer et al (2012 and2013a) in order to study undecalcified archaeological bone samples, especially when lesions were present. In addition to the improvements in the preparation of samples, other sophisticated microscope techniques were introduced to the study of archaeological remains-namely, the atomic force microscope (Thalhammer et al, 2001), the epifluorescence microscope, microscopic computerized tomography, and the confocal laser scanning microscope Rühli et al, 2007;Maggiano et al, 2009;Šefčáková et al, 2001).…”

Ancient pathogens in museal dry bone specimens: analysis of paleocytology and aDNA

“…In contrast to clinical practice, the use of histological stains in the field of anthropology is relatively rare [20,21]. Histological methods detecting past pathogens are applied to mummified specimens and old paraffin blocks of soft tissue [41].…”