Three-dimensional investigation and scoring of extracellular matrix remodeling during lung fibrosis using multiphoton microscopy
Three-dimensional investigation and scoring of extracellular matrix remodeling during lung fibrosis using multiphoton microscopy
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DOI：
10.1002/jemt.20400
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发表时间：
2007-02-01
影响因子：
2.5
通讯作者：
Schanne-Klein, Marie-Claire
中科院分区：
文献类型：
--
作者：
Pena, Ana-Maria;Fabre, Aurelie;Schanne-Klein, Marie-Claire
由于缺乏适当的方法，纤维化过程中胶原蛋白的组织特征很差。在这里，我们表明，多模态多光子显微镜提供了新的见解肺纤维化。我们在博莱霉素模型中表征了正常和纤维化肺组织，并表明纤维胶原的二次谐波产生揭示了纤维化的微米级三维空间分布。我们发现，未染色肺组织的双光子激发荧光和二次谐波成像的组合允许分离这种病理中的炎症和纤维化步骤，强调人特发性肺纤维化样本中成纤维细胞病灶的特征。最后，我们提出了肺纤维化的现象学评分，我们表明，他们明确地挑选出对照组和治疗组小鼠，在胸膜下区域具有更好的灵敏度和再现性。这些结果应该很容易推广到其他器官，作为一个准确的方法来评估纤维化过程中的细胞外基质重塑。
The organization of collagen during fibrotic processes is poorly characterized because of the lack of appropriate methodologies. Here we show that multimodal multiphoton microscopy provides novel insights into lung fibrosis. We characterize normal and fibrotic pulmonary tissue in the bleomycin model, and show that second-harmonic generation by fibrillar collagen reveals the micrometer-scale three-dimensional spatial distribution of the fibrosis. We find that combined two-photon excited fluorescence and second-harmonic imaging of unstained lung tissue allows separating the inflammatory and fibrotic steps in this pathology, underlining characteristic features of fibroblastic foci in human Idiopathic Pulmonary Fibrosis samples. Finally, we propose phenomenological scores of lung fibrosis and we show that they unambiguously sort out control and treated mice, with a better sensitivity and reproducibility in the subpleural region. These results should be readily generalized to other organs, as an accurate method to assess extracellular matrix remodeling during fibrosis.