Jasmine Singh, Taylor Nowlin, Gregory Seedorf, Steven Abman, Douglas Shepherd
Journal of Biomedical Optics, Vol. 22, Issue 07, 076011, (July 2017) https://doi.org/10.1117/1.JBO.22.7.076011
TOPICS: Retina, Microscopy, Tissue optics, Tissue clearing, 3D modeling, Oxygen, Luminescence, 3D image processing, 3D displays
Retinal vasculature develops in a highly orchestrated three-dimensional (3-D) sequence. The stages of retinal vascularization are highly susceptible to oxygen perturbations. We demonstrate that optical tissue clearing of intact rat retinas and light-sheet microscopy provides rapid 3-D characterization of vascular complexity during retinal development. Compared with flat mount preparations that dissect the retina and primarily image the outermost vascular layers, intact cleared retinas imaged using light-sheet fluorescence microscopy display changes in the 3-D retinal vasculature rapidly without the need for point scanning techniques. Using a severe model of retinal vascular disruption, we demonstrate that a simple metric based on Sholl analysis captures the vascular changes observed during retinal development in 3-D. Taken together, these results provide a methodology for rapidly quantifying the 3-D development of the entire rodent retinal vasculature.