Label-free photoacoustic microscopy for in-vivo tendon imaging using a fiber-based pulse laser

Tendons are tough, flexible, and ubiquitous tissues that connect muscle to bone. Tendon injuries are a common musculoskeletal injury, which affect 7% of all patients and are involved in up to 50% of sports-related injuries in the United States. Various imaging modalities are used to evaluate tendons, and both magnetic resonance imaging and sonography are used clinically to evaluate tendons with non-invasive and non-ionizing radiation. However, these modalities cannot provide 3-dimensional (3D) structural images and are limited by angle dependency. In addition, anisotropy is an artifact that is unique to the musculoskeletal system. Thus, great care should be taken during tendon imaging. The present study evaluated a functional photoacoustic microscopy system for in-vivo tendon imaging without labeling. Tendons have a higher density of type 1 collagen in a cross-linked triple-helical formation (65-80% dry-weight collagen and 1-2% elastin in a proteoglycan-water matrix) than other tissues, which provides clear endogenous absorption contrast in the near-infrared spectrum. Therefore, photoacoustic imaging with a high sensitivity to absorption contrast is a powerful tool for label-free imaging of tendons. A pulsed near-infrared fiber-based laser with a centered wavelength of 780 nm was used for the imaging, and this system successfully provided a 3D image of mouse tendons with a wide field of view (5 × 5 mm²).