Using a water-immersion optical objective in conjunction with a miniature 40-MHz ultrasonic transducer, we developed reflection-mode photoacoustic microscopy with a transverse resolution as high as 320 nm. Here, we further integrated two-photon microscopy capability into the system to enable multimodality in vivo biomedical imaging at submicron resolution. As a result, the system is capable of tri-modality label-free imaging of microvasculature, collagen, and cell morphology, based on the contrast of optical absorption, second-harmonic generation, and autofluorescence, respectively. In addition, we demonstrated simultaneous microscopic imaging of neuron and microvasculature in the brain cortex of a living mouse, which may offer new opportunities for studying the mechanisms of neurovascular coupling.
Liang Song, Wei Song, Yang Zhang, and Wei Zheng, "Fully integrated reflection-mode photoacoustic/two-photon microscopy in vivo
(Conference Presentation)," Proc. SPIE 9708, Photons Plus Ultrasound: Imaging and Sensing 2016, 97082H (Presented at SPIE BiOS: February 17, 2016; Published: 27 April 2016); https://doi.org/10.1117/12.2214685.4828179397001.
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