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Related Experiment Video

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3D Visualization of Retinal Vascular Pericytes in Mice by Immunostaining
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Quantifying three-dimensional rodent retina vascular development using optical tissue clearing and light-sheet

Jasmine N Singh1, Taylor M Nowlin2, Gregory J Seedorf2

  • 1University of Colorado Denver, Department of Physics, Denver, Colorado, United StatesbUniversity of Colorado Anschutz Medical Campus, Pediatric Heart Lung Center, Department of Pediatrics, Aurora, Colorado, United States.

Journal of Biomedical Optics
|July 19, 2017
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Summary

This study introduces a rapid 3-D imaging method for retinal vascular development using optical clearing and light-sheet microscopy. This technique accurately quantifies vascular complexity, even in disrupted retinal vasculature models.

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Area of Science:

  • Developmental Biology
  • Ophthalmology
  • Biomedical Imaging

Background:

  • Retinal vasculature development is a complex 3-D process.
  • Vascular development is sensitive to oxygen levels.
  • Current imaging methods for retinal vasculature have limitations.

Purpose of the Study:

  • To develop a rapid method for 3-D characterization of retinal vascular development.
  • To assess vascular complexity in intact retinas.
  • To provide a quantitative analysis of retinal vascular changes.

Main Methods:

  • Optical tissue clearing of intact rat retinas.
  • Light-sheet fluorescence microscopy for 3-D imaging.
  • Sholl analysis for quantifying vascular complexity.

Main Results:

  • Intact cleared retinas enable rapid 3-D imaging of retinal vasculature.
  • Light-sheet microscopy visualizes developmental changes without point scanning.
  • Sholl analysis effectively captures vascular changes in 3-D, including in disrupted models.

Conclusions:

  • This methodology offers rapid quantification of entire rodent retinal vasculature development in 3-D.
  • The technique is valuable for studying retinal vascular development and disruption.
  • It provides a significant advancement over traditional flat-mount preparations.