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.

Details

Title
Imaging of Murine Whole Lung Fibrosis by Large Scale 3D Microscopy aided by Tissue Optical Clearing
Author
Ochoa, Lorenzo F 1 ; Kholodnykh, Alexander 2 ; Villarreal, Paula 2 ; Tian, Bing 3 ; Pal, Rahul 2 ; Freiberg, Alexander N 4 ; Brasier, Allan R 5 ; Motamedi, Massoud 6 ; Vargas, Gracie 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 
 Center for Biomedical Engineering, The University of Texas Medical Branch, Galveston, USA 
 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 
 Department of Pathology, The University of Texas Medical Branch, Galveston, USA 
 Institute for Clinical and Translational Research, University of Wisconsin-Madison School of Medicine and Public Health, Madison, USA 
 Center for Biomedical Engineering, The University of Texas Medical Branch, Galveston, USA; Department of Ophthalmology, The University of Texas Medical Branch, Galveston, USA 
Pages
1-14
Publication year
2018
Publication date
Sep 2018
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2100362255
Copyright
© 2018. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.