Prof. Luis Dorfmann Profile

Prof. Luis Dorfmann

at Tufts Univ

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Proceedings Article | 4 March 2019 Presentation

Two-photon excited fluorescence properties of glycation-associated collagen crosslinks that are correlated with tissue stiffness (Conference Presentation)

Xi Yu, Christopher Flynn, Isaac Lasko, Sauro Liberatore, Luis Dorfmann, Irene Georgakoudi

Proceedings Volume 10890, 108901E (2019) https://doi.org/10.1117/12.2507617

KEYWORDS: Collagen, Tissues, Luminescence, Second-harmonic generation, Environmental sensing, Harmonic generation, Image resolution, Mathematical modeling, Cancer

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Collagen is a main constituent of the extracellular matrix, and its content, organization, and crosslinking status affect significantly tissue mechanical properties. Cells continuously sense their local mechanical environment via interactions that impact significantly cell fate and the development and progression of numerous diseases. Non-linear imaging, especially through the combined use of second harmonic generation (SHG) and two-photon excited fluorescence (TPEF), has the potential to provide detailed information regarding collagen organization, content, and crosslinking with sub-micron level resolution in a non-invasive and label-free manner. However, the crosslink fluorescence properties that are correlated with tissue stiffness aren’t well understood. Thus, we characterized the TPEF emission over 680 to 920 nm excitation and 400 to 780 nm emission of three types of collagen gels: a) plain collagen gels, b) collagen gels with a 250 mM ribose solution for five days prior to gelation, and c) following gelation. Uniaxial tensile testing of gels created using the same protocols was performed by a custom-built testing system. We identified two components, which peak at 440 nm and 510 nm, that were significantly correlated with the exponential stiffening detected in collagen gels treated with ribose following gelation. Thus, TPEF information acquired at these emission ranges can be used in future studies and in combination with SHG measurements to develop detailed mathematical models that aim to predict tissue micromechanical properties. The latter will be useful in helping us to understand mechanosensitive cell behaviors that ultimately dictate the progress of diseases such as fibrosis and cancer.

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