22 February 2018 Structured polarized light microscopy (SPLM) for mapping collagen fiber orientation of ocular tissues
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Glaucoma is a disease characterized by progressive and irreversible vision loss leading to blindness. This vision loss is believed to be largely determined by the biomechanics of the optic nerve head region. Optic nerve head biomechanics, in turn, is determined by the properties of the constituent collagen. However, it is challenging to visualize and quantify collagen morphology and orientation in situ, and therefore often studies of the region collagen have used histological sections. Here we describe SPLM, a novel imaging technique that combines structured light illumination and polarized light microscopy (PLM) to enable collagen fiber visualization and fiber orientation mapping without requiring tissue sectioning.

We developed a custom automated SPLM imaging system based on an upright microscope and a digital micromirror device (DMD) projector. The high spatial frequency patterns were used to achieve effective background suppression. Enhanced scattering sensitivity with SPLM resulted in images with highly improved visibility of collagen structures, even of tissues covered by pigment. SPLM produced improved fiber orientation maps from superficial layers compared to depth-averaged orientation from regular PLM. SPLM imaging provides valuable information of collagen fiber morphology and orientation in situ thus strengthening the study of ocular collagen fiber biomechanics and glaucoma.
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Bin Yang, Bin Yang, Bryn Brazile, Bryn Brazile, Ning-Jiun Jan, Ning-Jiun Jan, Andrew P. Voorhees, Andrew P. Voorhees, Ian A. Sigal, Ian A. Sigal, } "Structured polarized light microscopy (SPLM) for mapping collagen fiber orientation of ocular tissues ", Proc. SPIE 10546, Emerging Digital Micromirror Device Based Systems and Applications X, 105460I (22 February 2018); doi: 10.1117/12.2290133; https://doi.org/10.1117/12.2290133

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