Department of Anatomy and Cell Biology, Indiana University School of Medicine, Indianapolis, Indiana 46202, USA.
Microsc Res Tech. 2010 Mar;73(3):182-6. doi: 10.1002/jemt.20772.
Osteocytes are the most abundant of the bone cells. Each osteocyte is contained within its own lacuna and connected to adjacent osteocytes via fillipodial processes, which form an intricate network of canaliculi within the matrix. Studying this intricate network of cells and their processes is difficult, because it exists embedded within a densely mineralized matrix. Scanning electron microscopy (SEM) has been shown to be a useful tool for visualizing this cellular network, yet the techniques involved for preparing specimens has not been systematically explored. The goal of this study was to investigate how variations in acid-etching, both etching media and etching duration, affect SEM-based visualization of the osteocyte lacunar-canalicular network. Bone samples were embedded in plastic and then acid etched in either 9% (10, 20, 40, and 60 s durations) or 37% (5, 10, and 15 s) phosphoric acid. Specimens were imaged using SEM, and qualitative evaluation of the lacunar-canalicular network was undertaken. Our findings show acid etchingwith a 9% phosphoric acid solution for 20 s provided the most favorable visualization of the osteocyte lacunar-canalicular network.
成骨细胞是最丰富的骨细胞。每个成骨细胞都包含在自己的陷窝内，并通过丝状伪足与相邻的成骨细胞相连，这些丝状伪足在基质内形成一个错综复杂的小管网络。研究这个错综复杂的细胞网络及其过程是很困难的，因为它存在于一个高度矿化的基质中。扫描电子显微镜（SEM）已被证明是一种用于可视化这个细胞网络的有用工具，但用于制备标本的技术尚未得到系统的探索。本研究的目的是研究酸蚀刻（蚀刻介质和蚀刻时间）的变化如何影响基于 SEM 的骨细胞陷窝小管网络的可视化。将骨样本嵌入塑料中，然后用 9%（10、20、40 和 60 秒）或 37%（5、10 和 15 秒）的磷酸进行酸蚀刻。使用 SEM 对标本进行成像，并对陷窝小管网络进行定性评估。我们的发现表明，用 9%的磷酸溶液蚀刻 20 秒可以最有效地观察骨细胞陷窝小管网络。
