Dental Physical Sciences, Oral Growth and Development, Dental Institute, Barts' and The London School of Medicine and Dentistry, Queen Mary University of London, London E1 4NS, UK.
Eur Cell Mater. 2012 Jul 24;24:154-60; discussion 160-1. doi: 10.22203/ecm.v024a11.
Backscattered electron scanning electron microscopy (BSE SEM) is an invaluable method for studying the histology of the hard, mineralised components of poly-methyl methacrylate (PMMA) or other resin embedded skeletal and dental tissues. Intact tissues are studied in micro-milled or polished block faces with an electron-optical section thickness of the order of a half to one micron and with the area of the section as big as a whole--large or small--bone organ. However, BSE SEM does not give information concerning the distribution of uncalcified, 'soft', cellular and extracellular matrix components. This can be obtained by confocal microscopy of the same block and the two sorts of images merged but the blocks have to be studied in two microscope systems. The present work shows a new, simple and economic approach to visualising both components by using the triiodide ion in Lugol's iodine solution to stain the block surface prior to the application of any conductive coating--and the latter can be omitted if charging is suppressed by use of poor vacuum conditions in the SEM sample chamber. The method permits the use of archival tissue, and it will be valuable in studies of both normal growth and development and pathological changes in bones and joints, including osteoporosis and osteoarthritis, and tissue adaptation to implants.
背散射电子扫描电子显微镜（BSE-SEM）是一种非常有价值的方法，可用于研究聚甲基丙烯酸甲酯（PMMA）或其他树脂包埋的骨骼和牙齿组织的硬而矿化成分的组织学。完整的组织通过微铣或抛光块面进行研究，电子光学截面厚度约为半到一微米，截面面积与整个——大或小——骨骼器官一样大。然而，BSE-SEM 并不能提供关于未钙化的“软”细胞和细胞外基质成分分布的信息。这可以通过对同一块进行共聚焦显微镜检查并合并两种图像来获得，但块必须在两个显微镜系统中进行研究。本工作展示了一种新的、简单和经济的方法，可以通过在卢戈氏碘溶液中的三碘离子对块表面进行染色，来可视化这两种成分，而无需进行任何导电涂层——如果在 SEM 样品室中使用较差的真空条件来抑制电荷，则可以省略后者。该方法允许使用档案组织，它将在骨骼和关节的正常生长和发育以及病理变化的研究中非常有价值，包括骨质疏松症和骨关节炎，以及组织对植入物的适应。
