Macdonald J L, Kane A B
Lab Invest. 1986 Aug;55(2):177-85.
Identification of asbestos fibers and other mineral particles in tissues is important for the diagnosis of interstitial lung disease. Conventional procedures to identify mineral particles are applicable to tissue digests, homogenates, or thin sections prepared for transmission electron microscopy. Positive identification of mineral particles in these samples is achieved by energy dispersive x-ray analysis or crystalline diffraction patterns. These analytical techniques are difficult to use for identification of long, thin asbestos fibers within cells collected from effusions or by saline lavage. A new preparative procedure is presented which allows intracellular visualization of fibers in these samples. Mice were injected intraperitoneally with 100 micrograms of crocidolite asbestos. After 1 to 30 days, the free peritoneal cell population was collected by saline lavage and allowed to attach to Formvar/carbon coated grids in vitro. Cell spreading was induced by exposure to phorbol-12-myristate-13-acetate for an additional 4 hours. The flattened cells were fixed, dehydrated and air-dried before examination by transmission electron microscopy. This procedure allows direct visualization of intracellular fibers. The characteristic Fe and Si peaks of crocidolite asbestos were confirmed by energy dispersive x-ray analysis. This technique was used to study the kinetics of clearance of asbestos fibers from the free peritoneal macrophage population of mice.
鉴定组织中的石棉纤维和其他矿物颗粒对于间质性肺病的诊断很重要。鉴定矿物颗粒的传统方法适用于组织消化物、匀浆或为透射电子显微镜制备的薄片。通过能量色散X射线分析或晶体衍射图谱可对这些样品中的矿物颗粒进行阳性鉴定。这些分析技术难以用于鉴定从积液中收集的细胞内或通过盐水灌洗收集的细胞内的细长石棉纤维。本文介绍了一种新的制备方法，该方法可使这些样品中的纤维在细胞内可视化。给小鼠腹腔注射100微克青石棉。1至30天后，通过盐水灌洗收集游离腹膜细胞群，并使其在体外附着于福尔马弗/碳涂层网格上。通过暴露于佛波醇-12-肉豆蔻酸酯-13-乙酸酯4小时诱导细胞铺展。在通过透射电子显微镜检查之前，将扁平细胞固定、脱水并风干。该程序可直接观察细胞内纤维。通过能量色散X射线分析证实了青石棉的特征性铁和硅峰。该技术用于研究小鼠游离腹膜巨噬细胞群中石棉纤维清除的动力学。
