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

Sheldon J. J. Kwok; Ivan A. Kuznetsov; Moonseok Kim; Myunghwan Choi; Giuliano Scarcelli; Seok-Hyun Yun

doi:10.1117/12.2251565

24 April 2017 Selective two-photon collagen crosslinking in situ measured by Brillouin microscopy (Conference Presentation)

Sheldon J. J. Kwok, Ivan A. Kuznetsov, Moonseok Kim, Myunghwan Choi, Giuliano Scarcelli, Seok-Hyun Yun

Author Affiliations +

Proceedings Volume 10067, Optical Elastography and Tissue Biomechanics IV; 100670M (2017) https://doi.org/10.1117/12.2251565
Event: SPIE BiOS, 2017, San Francisco, California, United States

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.

Conference Presentation

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Sheldon J. J. Kwok, Ivan A. Kuznetsov, Moonseok Kim, Myunghwan Choi, Giuliano Scarcelli, and Seok-Hyun Yun "Selective two-photon collagen crosslinking in situ measured by Brillouin microscopy (Conference Presentation)", Proc. SPIE 10067, Optical Elastography and Tissue Biomechanics IV, 100670M (24 April 2017); https://doi.org/10.1117/12.2251565

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KEYWORDS

Tissues

Collagen

Microscopy

Confocal microscopy

In situ metrology

Cornea

Femtosecond phenomena

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