Second harmonic generation microscopy analysis of extracellular matrix changes in human idiopathic pulmonary fibrosis

Karissa B. Tilbury; James D. Hocker; Bruce L. Wen; Nathan Sandbo; Vikas Singh; Paul J. Campagnola

doi:10.1117/1.JBO.19.8.086014

18 August 2014 Second harmonic generation microscopy analysis of extracellular matrix changes in human idiopathic pulmonary fibrosis

Karissa B. Tilbury, James D. Hocker, Bruce L. Wen, Nathan Sandbo, Vikas Singh, Paul J. Campagnola

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Journal of Biomedical Optics, Vol. 19, Issue 8, 086014 (August 2014). https://doi.org/10.1117/1.JBO.19.8.086014

Abstract

Patients with idiopathic fibrosis (IPF) have poor long-term survival as there are limited diagnostic/prognostic tools or successful therapies. Remodeling of the extracellular matrix (ECM) has been implicated in IPF progression; however, the structural consequences on the collagen architecture have not received considerable attention. Here, we demonstrate that second harmonic generation (SHG) and multiphoton fluorescence microscopy can quantitatively differentiate normal and IPF human tissues. For SHG analysis, we developed a classifier based on wavelet transforms, principle component analysis, and a K-nearest-neighbor algorithm to classify the specific alterations of the collagen structure observed in IPF tissues. The resulting ROC curves obtained by varying the numbers of principal components and nearest neighbors yielded accuracies of <95% . In contrast, simpler metrics based on SHG intensity and collagen coverage in the image provided little or no discrimination. We also characterized the change in the elastin/collagen balance by simultaneously measuring the elastin autofluorescence and SHG intensities and found that the IPF tissues were less elastic relative to collagen. This is consistent with known mechanical consequences of the disease. Understanding ECM remodeling in IPF via nonlinear optical microscopy may enhance our ability to differentiate patients with rapid and slow progression and, thus, provide better prognostic information.

CC BY: © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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Karissa B. Tilbury, James D. Hocker, Bruce L. Wen, Nathan Sandbo, Vikas Singh, and Paul J. Campagnola "Second harmonic generation microscopy analysis of extracellular matrix changes in human idiopathic pulmonary fibrosis," Journal of Biomedical Optics 19(8), 086014 (18 August 2014). https://doi.org/10.1117/1.JBO.19.8.086014

Published: 18 August 2014

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