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Abstract
Pulmonary fibrosis, characterized by excessive collagen deposition in the lungs, comprises a key and debilitating component of chronic lung diseases. Methods are lacking for the direct visualization of fibrillar collagen throughout the whole murine lung, a capability that would aid the understanding of lung fibrosis. We combined an optimized organ-level optical clearing (OC) approach with large-scale, label-free multiphoton microscopy (MPM) and second harmonic generation microscopy (SHGM) to reveal the complete network of fibrillar collagen in whole murine lungs. An innate inflammation-driven model based on repetitive poly(I:C) challenge was evaluated. Following OC, mosaic MPM/SHGM imaging with 3D reconstruction and whole organ quantitative analysis revealed significant differences in collagen deposition between PBS and poly(I:C) treated lungs. Airway specific analysis in whole lung acquisitions revealed significant sub-epithelial fibrosis evident throughout the proximal conductive and distal airways with higher collagen deposition in the poly(I:C) group vs PBS group. This study establishes a new, powerful approach based on OC and MPM/SHGM imaging for 3D analysis of lung fibrosis with macroscopic views of lung pathology based on microscopy and providing a new way to analyze the whole lung while avoiding regional sampling bias.
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1 Center for Biomedical Engineering, The University of Texas Medical Branch, Galveston, USA; Department of Neuroscience, Cell Biology and Anatomy, The University of Texas Medical Branch, Galveston, USA
2 Center for Biomedical Engineering, The University of Texas Medical Branch, Galveston, USA
3 Department of Internal Medicine, The University of Texas Medical Branch, Galveston, USA; Sealy Center for Molecular Medicine, The University of Texas Medical Branch, Galveston, USA
4 Department of Pathology, The University of Texas Medical Branch, Galveston, USA
5 Institute for Clinical and Translational Research, University of Wisconsin-Madison School of Medicine and Public Health, Madison, USA
6 Center for Biomedical Engineering, The University of Texas Medical Branch, Galveston, USA; Department of Ophthalmology, The University of Texas Medical Branch, Galveston, USA