Vascular retinopathy is a leading cause of preventable blindness in diabetes. While most studies focus on individual neural or vascular structures, there remains a knowledge gap in understanding the retinal neurovascular interactions due to injury and repair. We seek to demonstrate 3-D neurovascular interactions with a multi-view light-sheet imaging system. Intact 3-D retinas from 2-month-old mice were fixed and permeabilized followed by the optical clearance. Retinal vessels and neuron cells were stained with fluorescent labels. After refractive index matching, multi-view light-sheet imaging was performed at different angles followed by 3-D registration and reconstruction. This enabled 3-D co-registration of the vasculature in relation to the neuron cells for virtualization. Therefore, the 3-D neuro-microvascular network could be revealed with both deep tissue penetration and high spatial resolution for investigating hyperoxia and diabetes-associated vascular retinopathy.
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