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

Jasmine N. Singh, Taylor M. Nowlin, Gregory J. Seedorf, Steven H. Abman, Douglas P. Shepherd

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

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.

Original languageEnglish (US)
Article number076011
JournalJournal of biomedical optics
Volume22
Issue number7
DOIs
StatePublished - Jul 1 2017

Keywords

  • fluorescence
  • image acquisition/recording
  • image analysis
  • microscopy

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Atomic and Molecular Physics, and Optics
  • Biomedical Engineering

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