Ocular chemical damage may induce limbal vessel ischemia and neovascularization, but the pathophysiology of the disease is not completely known. To observe changes in blood vessels after alkaline burn, we monitored the anterior segment and choroidal vasculature using a photoacoustic microscope (OR-PAM). We were able to observe not only the iris blood vessels but also the choroidal vessels under the sclera, which were difficult to be observed with conventional photographs. After alkali burning, we observed neovascularization and limbal ischemia and successfully tracked changes in vasculature during the 7-day healing process. We also used the RANdom SAmple Consensus (RANSAC) method to segment the abnormally generated blood vessels in the cornea by detecting the eyeball surface and successfully visualize the distance from each PA signal to the center of the eye. We believe that photoacoustic imaging has an important potential to reveal the pathophysiology of limb ischemia and neovascularization.
Photoacoustic microscopy (PAM) provides high resolution and large penetration depth by utilizing the high optical sensitivity and low scattering of ultrasound. Hybrid PAM systems can be classified into two categories: opticalresolution photoacoustic microscopy (OR-PAM) and acoustic-resolution photoacoustic microscopy (AR-PAM). ORPAM provides a very high lateral resolution with a strong optical focus, but the penetration depth is limited to one optical transport mean free path. AR-PAM provides a relatively greater penetration depth using diffused light in biological tissues. The resolution of AR-PAM is determined by its ultrasonic parameters. In this study, we performed an in vivo testing of a switchable OR-/AR-PAM system. In this system, two modes can be switched by changing its collimator lens and optical fiber. The lateral resolution of OR-PAM was measured using a resolution test target, and the full width at half maximum (FWHM) of the edge spread function was 2.5 μm. To calculate the lateral resolution of ARPAM, a 6-μm-diameter carbon fiber was used, and the FWHM of the line spread function was 80.2 μm. We successfully demonstrated the multiscale imaging capability of the switchable OR-/AR-PAM system by visualizing microvascular networks in mouse ears, brain, legs, skin, and eyes.