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27 February 2019Combined speckle variance optical coherence tomography and multiphoton microscopy for in vivo chick CAM imaging
Combining optical coherence tomography (OCT) and multiphoton microscopy (MPM) can provide multimodal imaging of the microstructure of biological tissues. As a functional extension of conventional OCT, speckle variance OCT (SVOCT) can be applied to image microvasculature to improve the blood vessel visualization. In this paper, a combined SVOCT and MPM system is developed to visualize the chorioallantoic membrane (CAM) of a chick embryo, which contains extensive blood vessel network. Based on the different temporal decorrelation characteristics of the fluid flow and the surrounding stationary structure, SV-OCT enables enhanced contrast of fluid flow from the surrounding structure. As a result, SV-OCT can achieve detailed mapping of the CAM microvasculature at the tissue level. Meanwhile, MPM enables vascular imaging at the cellular level, where two-photon excitation fluorescence (TPEF) images fluorescein dye injected into the blood stream, and second harmonic generation (SHG) visualizes the collagen fiber structures in the vessel wall and the surrounding tissues. Therefore, the combined SV-OCT and MPM system provides complementary information about the microvasculature structures in the chick CAM. The combined system is shown to be a powerful tool for interpreting the microvasculature, by allowing the visualization of the blood vessel network in a relatively large field of view at the tissue level with SV-OCT, and by providing cellular-level information in local regions of interest with MPM.