Laboratory of Histology and Embryology, Medical School, Athens University, Athens, Greece.
Histopathology. 2012 Dec;61(6):1174-82. doi: 10.1111/j.1365-2559.2012.04335.x. Epub 2012 Sep 7.
The aim of this study was the development of a simplified technique for bone mineral histomorphology on large undecalcified bone samples. Established techniques, such as undecalcified bone thin sectioning, ultrathin grinding, surface-stained block grinding and micro-computerized tomography (CT), are expensive, time-consuming and put very high demands on equipment, safety standards, personnel and laboratory facilities.
The method is based on the surface-stained block-grinding principle; however, its novelty lies in the selection of user-friendly, safe and low-cost materials, equipment and digitization techniques. We describe in detail the relevant steps, as well as many practical tips for their successful implementation: accurate bone cutting in thin sections with a customized arrangement on a commercial bandsaw, defatting with sodium hypochlorite, embedding in epoxy resin blocks at room temperature, silicon carbide paper grinding, von Kossa staining, flatbed scanner digitization and image processing.
We believe that the proposed methodology could contribute to the expansion of the study of bone tissue, as it enables the rapid examination of bone specimens on a large scale with minimal laboratory requirements and consumables costs.
本研究旨在开发一种简化的大型非脱钙骨样本骨组织形态计量学技术。传统技术,如非脱钙骨薄片切割、超薄磨片、表面染色块研磨和微计算机断层扫描(CT),既昂贵又耗时,对设备、安全标准、人员和实验室设施要求很高。
该方法基于表面染色块研磨原理;然而,其新颖之处在于选择了用户友好、安全且低成本的材料、设备和数字化技术。我们详细描述了相关步骤以及许多成功实施的实用技巧:在商用带锯上定制排列以进行精确的薄片骨切割、次氯酸钠脱脂、室温下包埋在环氧树脂块中、碳化硅砂纸研磨、von Kossa 染色、平板扫描仪数字化和图像处理。
我们相信,所提出的方法可以为骨组织研究的扩展做出贡献,因为它可以在最小的实验室要求和耗材成本下快速大规模检查骨标本。