Mühlfeld Christian, Knudsen Lars, Ochs Matthias
Institute of Functional and Applied Anatomy, Hannover Medical School, Hannover, Germany.
Methods Mol Biol. 2013;931:367-90. doi: 10.1007/978-1-62703-056-4_18.
This chapter deals with the stereological quantification of structural characteristics of the lung. The aim of design-based stereological methods is the unbiased and efficient estimation of structural features without making any assumptions on the underlying nature of the biological sample. The methods are based on rigorous sampling of location and orientation, the application of appropriate test systems, and the controlling of the precision of the estimates. Here, we describe the workflow from the fixation of the lung over the processing of the tissue samples to gaining estimates on the structural properties of the lung. Specifically, this chapter deals with methods for estimating the reference volume, sampling location, and sampling orientation, estimating volumes and surface areas of alveolar compartments, estimating total alveolar number, performing stereology at light and electron microscopic level, and dealing with technical problems such as tissue shrinkage. The procedures are illustrated using a worked example from the authors' own laboratory.
本章论述肺结构特征的体视学定量分析。基于设计的体视学方法的目的是在不对生物样本的潜在性质做任何假设的情况下,无偏且高效地估计结构特征。这些方法基于对位置和取向的严格抽样、合适测试系统的应用以及对估计精度的控制。在此,我们描述从肺的固定到组织样本处理,再到获得肺结构特性估计值的工作流程。具体而言,本章涉及估计参考体积、抽样位置和抽样取向的方法,估计肺泡腔的体积和表面积,估计肺泡总数,在光镜和电镜水平进行体视学分析,以及处理诸如组织收缩等技术问题。文中使用作者自己实验室的一个实例对这些程序进行了说明。