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Pelvic organ prolapse (POP) is a gynecological disorder described by the descent of superior pelvic organs into or out of the vagina as a consequence of disrupted muscles and tissue. A thorough understanding of the etiology of POP is limited by the availability of clinically relevant samples, restricting longitudinal POP studies on soft-tissue biomechanics and structure to POP-induced models such as fibulin-5 knockout (FBLN5^-/-) mice. Despite being a principal constituent in the extracellular matrix, little is known about structural perturbations to collagen networks in theFBLN5^-/-mouse cervix. We identify significantly different collagen network populations in normal and prolapsed cervical cross-sections using two label-free, nonlinear microscopy techniques. Collagen in the prolapsed mouse cervix tends to be more isotropic, and displays reduced alignment persistence via 2-D Fourier transform analysis of images acquired using second harmonic generation microscopy. Furthermore, coherent Raman hyperspectral imaging revealed elevated disorder in the secondary structure of collagen in prolapsed tissues. Our results underscore the need forin situmultimodal monitoring of collagen organization to improve POP predictive capabilities.

Confocal microscopy provides optical sectioning that is invaluable for many applications, most notably imaging into thick samples. However, the high cost of commercial confocal microscopes limits uses to specialized research and clinical settings. We present a minimalistic line-scanning confocal microscope costing less than $6,500 with optical performance comparable to a commercial laser scanning system. The optical sectioning and imaging performance are shown through measurement of the axial line-spread function and imaging of biological samples of varying thickness. Comparison is made to commercial widefield and confocal microscopes. The low cost of goods and optical sectioning capability of this microscope will allow the use of confocal microscopy in additional research and educational settings.