Second Harmonic Generation analysis of the ECM in idiopathic pulmonary fibrosis Idiopathic pulmonary fibrosis (IPF) is a disorder characterized by unrelenting scarring and stiffening of the lungs that leads to the death of an estimated 34,000 individuals in the U.S. each year. Unfortunately, individuals with IPF have extremely limited treatment options, as no effective drugs have been identified to halt the progression of fibrosis. Despite the importance of collagens to the structural organization both normal and remodeled ECM, little is known about how collagen structure in IPF differs from that of normal tissue architecture. There is a clear need to develop highly specific/sensitive techniques to probe collagen structure and organization in IPF tissues. In this project we will implement new collagen specific analyses using the high resolution microscopy technique of Second Harmonic Generation (SHG). This method is sensitive to both the fibrillar organization and also sub-resolution aspects of macro and supramolecular assembly. Here we will utilize SHG microscopy to: 1) determine the how pathologic collagen organization (seen in IPF) differs from normal tissue; 2) identify and quantify areas of active fibrosis (enriched in collagen III) from old or mature fibrosis (high in collagen I) in IPF lung specimens; 3) assess changes in elastin and collagen distribution during disease progression; and 4) correlate areas of high collagen III/I signal in IPF histologic samples with clinical markers of disease activity. As part of the project, we will develop customized automated machine vision routines to automatically classify tissues in terms of severity. We will specifically focus all of our efforts on studying structure around fibroblastic foci, which will be identified by other microscope modalities. These foci are thought to be at the leading edge of ECM remodeling but the dynamics of their formation in relationship to the overall fibrotic process remain unclear. We hypothesize that these structural changes will serve as label- free biomarkers of IPF and further hypothesize that the collagen is altered specifically around foci in a manner which is associated with disease progression. The information gained may form the basis of future prognostic/diagnostic schemes. We propose 2 Aims:
Aim 1 Polarization resolved SHG to determine distribution of Col I/III and other ECM changes in different stages of IPF.
Aim 2. Develop classification system of morphological changes in IPF visualized by SHG.

Public Health Relevance

Second Harmonic Generation analysis of the ECM in idiopathic pulmonary fibrosis Narrative Idiopathic fibrosis (IPF) patients have poor survival rates and there is also a lack of diagnostic/prognostic tools that have sufficient sensitivity and specificity to evaluate changes in collagen in the extracellular matrix. The methods developed here will improve upon these limitations and may lay the groundwork for eventual non- invasive in vivo imaging.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Exploratory/Developmental Grants (R21)
Project #
Application #
Study Section
Enabling Bioanalytical and Imaging Technologies Study Section (EBIT)
Program Officer
Harabin, Andrea L
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Wisconsin Madison
Biomedical Engineering
Biomed Engr/Col Engr/Engr Sta
United States
Zip Code
Campbell, Kirby R; Wen, Bruce; Shelton, Emily M et al. (2017) 3D second harmonic generation imaging tomography by multi-view excitation. Optica 4:1171-1179
Wen, Bruce; Campbell, Kirby R; Tilbury, Karissa et al. (2016) 3D texture analysis for classification of second harmonic generation images of human ovarian cancer. Sci Rep 6:35734
Kim, Hyunwoo J; Xu, Jia; Vemuri, Baba C et al. (2015) Manifold-valued Dirichlet Processes. Proc Int Conf Mach Learn 2015:1199-1208
Kim, Hyunwoo J; Adluru, Nagesh; Banerjee, Monami et al. (2015) Interpolation on the manifold of K component GMMs. Proc IEEE Int Conf Comput Vis 2015:2884-2892