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Proceedings Paper

Selective two-photon collagen crosslinking in situ measured by Brillouin microscopy (Conference Presentation)

Paper Abstract

Two-photon polymerization and crosslinking are commonly used methods for microfabrication of three-dimensional structures with applications spanning from photonic microdevices, drug delivery systems, to cellular scaffolds. However, the use of two-photon processes for precise, internal modification of biological tissues has not yet been reported. One of the major challenges has been a lack of appropriate tools to monitor and characterize crosslinked regions nondestructively. Here, we demonstrate spatially selective two-photon collagen crosslinking (2P-CXL) in intact tissue for the first time. Using riboflavin photosensitizer and femtosecond laser irradiation, we crosslinked a small volume of tissue within animal corneas. Collagen fiber orientations and photobleaching were characterized by second harmonic generation and two-photon fluorescence imaging, respectively. Using confocal Brillouin microscopy, we measured local changes in longitudinal mechanical moduli and visualized the cross-linked pattern without perturbing surrounding non-irradiated regions. 2P-CXL-induced tissue stiffening was comparable to that achieved with conventional one-photon CXL. Our results demonstrate the ability to selectively stiffen biological tissue in situ at high spatial resolution, with broad implications in ophthalmology, laser surgery, and tissue engineering.

Paper Details

Date Published: 24 April 2017
PDF: 1 pages
Proc. SPIE 10067, Optical Elastography and Tissue Biomechanics IV, 100670M (24 April 2017); doi: 10.1117/12.2251565
Show Author Affiliations
Sheldon J. J. Kwok, Massachusetts General Hospital (United States)
Harvard Medical School (United States)
Massachusetts Institute of Technology (United States)
Ivan A. Kuznetsov, Massachusetts General Hospital (United States)
Johns Hopkins Univ. (United States)
Moonseok Kim, Massachusetts General Hospital (United States)
Myunghwan Choi, Massachusetts General Hospital (United States)
Sungkyunkwan Univ. (Korea, Republic of)
Giuliano Scarcelli, Massachusetts General Hospital (United States)
Univ. of Maryland, College Park (United States)
Seok-Hyun Yun, Massachusetts General Hospital (United States)
Harvard Medical School (United States)
Massachusetts Institute of Technology (United States)


Published in SPIE Proceedings Vol. 10067:
Optical Elastography and Tissue Biomechanics IV
Kirill V. Larin; David D. Sampson, Editor(s)

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