Blindness is a major public health issue globally. It has been reported that retinal diseases are the most and second most cause of irreversible blindness in developed and developing countries. Although it has been proven that structural and functional alterations of retina are associated with a variety of ocular disorders, the pathogenesis is still unclear. In vivo longitudinal imaging of retina at subcellular resolution could provide a powerful tool in discovering the progression of retinal diseases and biological basis. Since mouse models of human diseases have been developed rapidly, in vivo imaging of the mouse retina is now considered an ideal approach to study the pathogenesis of retinal disorders. The mouse eye has unique optical property of larger (~0.5) numerical aperture (NA)...[ Read more ]

Blindness is a major public health issue globally. It has been reported that retinal diseases are the most and second most cause of irreversible blindness in developed and developing countries. Although it has been proven that structural and functional alterations of retina are associated with a variety of ocular disorders, the pathogenesis is still unclear. In vivo longitudinal imaging of retina at subcellular resolution could provide a powerful tool in discovering the progression of retinal diseases and biological basis. Since mouse models of human diseases have been developed rapidly, in vivo imaging of the mouse retina is now considered an ideal approach to study the pathogenesis of retinal disorders. The mouse eye has unique optical property of larger (~0.5) numerical aperture (NA) compared with that of human eye (~0.2), which provides two-photon fluorescence microscopy with high resolution of subcellular level. However, the major challenge is the large optical aberration of mouse eye which blurs the excitation laser focus and lowers the imaging resolution.

In this work, we advance adaptive optics (AO) in two-photon microscopy to correct the mouse eye aberration and use this technology to in vivo visualize the progression of retinal diseases in mouse model. Specifically, we firstly proposed a robust and faster method for calibration of the deformable mirror and the wavefront sensor based on the Michelson interferometer, which improves the calibration efficiency and accuracy. Next, we did in vivo morphological and functional imaging of mouse retina with adaptive optics two-photon microscopy and showed the dynamic changes of vascular structures and flow speed in response to NMDA. My thesis work examined and optimized the capability of this technology for the in vivo study of the dynamic progression of retinal disease in mouse models.