Quantifying three-dimensional rodent retina vascular development using optical tissue clearing and light-sheet microscopy

J Biomed Opt. 2017 Jul 1;22(7):76011. doi: 10.1117/1.JBO.22.7.076011.

Abstract

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.

MeSH terms

  • Animals
  • Microscopy, Fluorescence*
  • Rats
  • Retina / embryology*
  • Retinal Vessels / embryology*