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Author:
Date:
2016-06-20
[Abstract] Although it is possible to use a tartrate-resistant acid phosphatase (TRAP) stain to assist in identifying osteoclasts, a separate method is needed to determine the bone resorption activity of osteoclasts. Since osteoclasts leave “pits” after bone matrix resorption (Charles et al., 2014), it is possible to stain pits as a method of measuring osteoclast bone resorption activity. The pit assay protocol enables researchers to stain bony slices that were co-cultured with osteoclasts with toluidine blue in order to allow the visualization, capture, and analysis of osteoclast resorptive activity based on the number, size and depth of pits (Zhou et al., 2015). The pit assay protocol is separated into three sequential stages: Preparation of bone slices (1); preparation of ...
[摘要] 尽管可以使用抗酒石酸盐的酸性磷酸酶（TRAP）染色来辅助鉴定破骨细胞，但是需要单独的方法来确定破骨细胞的骨吸收活性。 由于破骨细胞在骨基质吸收之后离开"凹陷"（Charles等人，2014），因此可以将斑点染色作为测量破骨细胞骨吸收活性的方法。 坑测定方案使研究人员能够将与破骨细胞与甲苯胺蓝共培养的骨切片染色，以便基于坑的数量，尺寸和深度可视化，捕获和分析破骨细胞再吸收活性（Zhou等人， et al。，2015）。 坑测定方案分为三个连续阶段：骨切片的制备（1）; 制备破骨细胞前体（Ross等人，2006; Teitelbaum等人，2000）（2）和骨吸收坑测定（3）。
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