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Proceedings Paper

Hierarchical model of fibrillar collagen distribution for polarization-resolved SHG microscopy
Author(s): Adam E. Tuer; Margarete K. Akens; Serguei Krouglov; Daaf Sandkuijl; Brian C. Wilson; Cari M. Whyne; Virginijus Barzda
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Paper Abstract

A hierarchical model of the organization of fibrillar collagen is developed and its implications on polarization-resolved second harmonic generation (SHG) microscopy are investigated. A “ground-up” approach is employed to develop the theory for understanding of the origin of SHG from fibrillar collagen. The effects of fibril ultrastructure and fibril macroscopic organization on the second-order polarization properties of fibrillar collagen are presented in conjunction with recent ab initio results performed on a collagen triple-helix model (-GLY-PRO-HYP-)n. Various tissues containing fibrillar collagen are quantified using a polarization-resolved SHG technique, termed polarization-in, polarization-out (PIPO) and interpreted in light of the aforementioned theory. The method involves varying the incident laser polarization, while monitoring the SHG intensity through an analyzer. From the SHG polarization data the orientation of the fibers, in biological tissue, can be deduced. Unique PIPO signatures are observed for different rat tissues and interpreted in terms of the collagen composition, fibril ultrastructure, and macroscopic organization. Similarities and discrepancies in the second-order polarization properties of different collagen types and ultrastructures will be presented. PIPO SHG microscopy shows promise in its ability to quantify the organization of collagen in various tissues. The ability to characterize the structure of collagen in various tissue microenvironments will aid in the study of numerous collagen related biological process, including tissue diseases, wound repair, and tumor development and progression.

Paper Details

Date Published: 22 February 2013
PDF: 8 pages
Proc. SPIE 8588, Multiphoton Microscopy in the Biomedical Sciences XIII, 85881O (22 February 2013); doi: 10.1117/12.2000724
Show Author Affiliations
Adam E. Tuer, Univ. of Toronto (Canada)
Margarete K. Akens, Sunnybrook Health Sciences Ctr. (Canada)
Serguei Krouglov, Univ. of Toronto (Canada)
Daaf Sandkuijl, Univ. of Toronto (Canada)
Brian C. Wilson, Ontario Cancer Institute (Canada)
Cari M. Whyne, Sunnybrook Health Sciences Ctr. (Canada)
Virginijus Barzda, Univ. of Toronto (Canada)


Published in SPIE Proceedings Vol. 8588:
Multiphoton Microscopy in the Biomedical Sciences XIII
Ammasi Periasamy; Karsten König; Peter T. C. So, Editor(s)

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