Unsupervised determination of lung tumor margin with widefield polarimetric second-harmonic generation microscopy

Abstract

The extracellular matrix (ECM) is amongst many tissue components affected by cancer, however, morphological changes of the ECM are not well-understood and thus, often omitted from diagnostic considerations. Polarimetric second-harmonic generation (P-SHG) microscopy allows for visualization and characterization of collagen ultrastructure in the ECM, aiding in better understanding of the changes induced by cancer throughout the tissue. In this paper, a large region of hematoxylin and eosin-stained human lung section, encompassing a tumor margin, connecting a significant tumor portion to normal tissue was imaged with P-SHG microscopy. The resulting polarimetric parameters were utilized in principal components analysis and unsupervised k-means clustering to separate normal- and tumor-like tissue. Consequently, a pseudo-color map of the clustered tissue regions is generated to highlight the irregularity of the ECM collagen structure throughout the region of interest and to identify the tumor margin.

Polarimetric response of second harmonic generation in microscopy of chiral fibrillar structures

“…These organizations can be decomposed into several basic fiber configurations affecting polarimetric SHG microscopy results, namely parallel/antiparallel fibers that can be tilted out of the image plane and crossed in 2D or 3D configuration. The ultrastructural information about the fibrillar structures obtained from the polarimetric SHG imaging can be used to study, for example, changes in collagenous tissue due to tumor development, wound healing, fibrosis, and aging (28,(54)(55)(56)(57)(58)(59).…”