Fourier transform-second harmonic generation (FT-SHG) imaging is used as a technique for evaluating collagenase-induced injury in horse tendons. The differences in collagen fiber organization between normal and injured tendon are quantified. Results indicate that the organization of collagen fibers is regularly oriented in normal tendons and randomly organized in injured tendons. This is further supported through the use of additional metrics, in particular, the number of dark (no/minimal signal) and isotropic (no preferred fiber orientation) regions in the images, and the ratio of forward-to-backward second-harmonic intensity. FT-SHG microscopy is also compared with the conventional polarized light microscopy and is shown to be more sensitive to assessing injured tendons than the latter. Moreover, sample preparation artifacts that affect the quantitative evaluation of collagen fiber organization can be circumvented by using FT-SHG microscopy. The technique has potential as an assessment tool for evaluating the impact of various injuries that affect collagen fiber organization.

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We present three-dimensional Fourier transform-second-harmonic generation (3D FT-SHG) imaging, a generalization of the previously reported two-dimensional FT-SHG, to quantify collagen fiber organization from 3D image stacks of biological tissues. The current implementation calculates 3D preferred orientation of a region of interest, and classifies regions of interest based on orientation anisotropy and average voxel intensity. Presented are some example applications of the technique which reveal the layered structure of collagen fibers in porcine sclera, and estimates the cut angle of porcine tendon tissues. This technique shows promising potential for studying biological tissues that contain fibrillar structures in 3D.

A new class of optical oxygen sensor that can be photopatternable by traditional UV lithography is presented. They are fabricated using photopatternable spin-on silicone (polydimethyl-siloxane, PDMS) with oxygen sensitive luminescent dyes. It has a good adhesion property and can be applied on glass or on photopolymer (SU-8) without any additional surface treatments. The optimum mixture composition for patternable oxygen sensitive membranes is investigated and its optical properties are characterized. Proof-of-concepts for two applications, intensity-based oxygen sensing with SU-8 based structure and self-calibration fluidic oxygen sensor, are described. These photopatternable optical membranes will find many applications wherever small patterns of oxygen sensitive membranes are required.