Utrecht Institute for Pharmaceutical Sciences, Utrecht Univ., The Netherlands.
Am J Physiol Lung Cell Mol Physiol. 2010 Dec;299(6):L843-51. doi: 10.1152/ajplung.00192.2010. Epub 2010 Oct 8.
Emphysema is characterized by enlargement of the alveoli, which is the most important parameter to assess the presence and severity of this disease. Alveolar enlargement is primarily defined on morphological criteria; therefore, characterization of this disease with morphological parameters is a prerequisite to study the pathogenesis. For this purpose, different methods of lung fixation were evaluated in a murine model of LPS-induced lung emphysema. Five different methods of lung fixation were evaluated: intratracheal instillation of fixatives, in situ fixation, fixed-volume fixation, vascular whole body perfusion, and vacuum inflation. In addition, the effects of three different fixatives (10% formalin, Carnoy's, and agarose/10% formalin solution) and two embedding methods (paraffin and plastic) were investigated on the murine lung morphology. Mice received intranasal administration of LPS to induce alveolar wall destruction. Quantification of air space enlargement was determined by mean linear intercept analysis, and the histological sections were analyzed for the most optimal fixation method. Additionally, routine immunohistological staining was performed on lung tissue of PBS-treated mice. Intratracheal instillation of formalin or agarose/formalin solution, in situ fixation, and fixed-volume fixation provided a normal lung architecture, in contrast to the lungs fixed via whole body perfusion and vacuum inflation. Formalin-fixed lungs resulted in the most optimal lung morphology for lung emphysema analysis when embedded in paraffin, while for Carnoy's fixed lungs, plastic embedding was preferred. The histological findings, the mean linear intercept measurement, and the immunohistochemistry data demonstrated that fixation by intratracheal instillation of 10% formalin or in situ fixation with 10% formalin are the two most optimal methods to fix lungs for alveolar enlargement analysis to study lung emphysema.
肺气肿的特征是肺泡扩大，这是评估这种疾病存在和严重程度的最重要参数。肺泡扩大主要根据形态学标准来定义；因此，用形态学参数来描述这种疾病是研究发病机制的前提。为此，在 LPS 诱导的肺肺气肿小鼠模型中评估了五种不同的肺固定方法。评估了五种不同的肺固定方法：气管内滴注固定剂、原位固定、固定体积固定、血管全身灌注和真空膨胀。此外，还研究了三种不同的固定剂（10%甲醛、卡诺氏液和琼脂糖/10%甲醛溶液）和两种包埋方法（石蜡和塑料）对小鼠肺形态的影响。小鼠接受鼻内 LPS 给药以诱导肺泡壁破坏。通过平均线性截距分析确定气腔扩大的定量，并用组织学切片分析最佳固定方法。此外，还对 PBS 处理的小鼠肺组织进行了常规免疫组织化学染色。与全身灌注和真空膨胀固定的肺相比，气管内滴注甲醛或琼脂糖/甲醛溶液、原位固定和固定体积固定提供了正常的肺结构。当石蜡包埋时，甲醛固定的肺为肺气肿分析提供了最佳的肺形态，而对于卡诺氏固定的肺，更倾向于使用塑料包埋。组织学发现、平均线性截距测量和免疫组织化学数据表明，气管内滴注 10%甲醛或原位固定 10%甲醛是固定肺以进行肺泡扩大分析以研究肺气肿的两种最佳方法。
