Xiao Rui, Goldklang Monica P, D'Armiento Jeanine M
Department of Anesthesiology, College of Physicians and Surgeons, Columbia University, New York, New York.
Am J Respir Cell Mol Biol. 2016 Nov;55(5):708-715. doi: 10.1165/rcmb.2016-0143OC.
Lung morphometry was introduced over 50 years ago to provide quantitative evaluation of the lung structure. The existing parameters, such as mean linear intercept and destructive index, suffer from simplistic data interpretation and a subjective data acquisition process. To overcome these existing shortcomings, parenchymal airspace profiling (PAP) was developed to provide a more detailed and unbiased quantitative method. Following the standard protocols of fixation, embedding, and sectioning, lung micrographs were: (1) marked with nonparenchymal area, preprocessed, and binarized under the researcher's supervision; (2) analyzed with a statistical learning method, Gaussian mixture model, to provide an unbiased categorization of parenchymal airspace compartments, corresponding to a single alveolus, alveolar sac, and ductal/destructive airspace; and (3) further quantified into morphometric parameters, including reference volume, alveolar count, and ductal/destructive fraction (DF) based on stereological principles. PAP was performed on hematoxylin and eosin-stained lung sections from mice and rabbits. Unbiased categorization revealed differences in alveolar size among several mouse strains (NZW/LacJ<AKR/J<A/J<C57BL/6J) and across species (mouse<rabbit). Further quantification indicates that parenchymal destruction, modeled in mouse lungs with 1-month smoke exposure, resulted in decreased alveolar count, increased DF, but no significant differences in mean linear intercept. DF also provides a robust measurement that is not biased by processing artifacts, magnification, or reference volume, which are common limitations in human lung biopsies or data obtained from different laboratories. PAP is a novel approach to lung morphometry that offers more detailed characterization of the lung structure, sensitivity, and robustness than presently used methods for evaluating parenchymal destruction.
肺形态测定法在50多年前就已引入,用于对肺结构进行定量评估。现有的参数,如平均线性截距和破坏指数,存在数据解释过于简单以及数据采集过程主观的问题。为克服这些现有缺点,开发了实质气腔剖析(PAP)法,以提供一种更详细且无偏倚的定量方法。按照固定、包埋和切片的标准方案,对肺显微图像进行如下处理:(1)在研究人员的监督下标记非实质区域、预处理并进行二值化处理;(2)采用统计学习方法高斯混合模型进行分析,以对实质气腔隔室进行无偏倚分类,分别对应单个肺泡、肺泡囊以及导管/破坏气腔;(3)根据体视学原理进一步量化为形态计量学参数,包括参考体积、肺泡计数以及导管/破坏分数(DF)。PAP法应用于小鼠和兔子的苏木精-伊红染色肺切片。无偏倚分类揭示了几种小鼠品系(NZW/LacJ<AKR/J<A/J<C57BL/6J)之间以及不同物种(小鼠<兔子)之间肺泡大小的差异。进一步的量化表明,在小鼠肺中模拟1个月的烟雾暴露导致实质破坏,表现为肺泡计数减少、DF增加,但平均线性截距无显著差异。DF还提供了一种可靠的测量方法,不受处理伪像、放大倍数或参考体积的影响,而这些是人类肺活检或从不同实验室获得的数据中常见的局限性。PAP是一种用于肺形态测定的新方法,与目前用于评估实质破坏的方法相比,它能更详细地表征肺结构、具有更高的敏感性和更强的稳健性。