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

doi:10.1117/1.JBO.22.7.076011

18 July 2017 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

Author Affiliations +

Journal of Biomedical Optics, Vol. 22, Issue 7, 076011 (July 2017). https://doi.org/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.

Citation Download Citation

Jasmine N. Singh, Taylor M. Nowlin, Gregory J. Seedorf, Steven H. Abman, and Douglas P. Shepherd "Quantifying three-dimensional rodent retina vascular development using optical tissue clearing and light-sheet microscopy," Journal of Biomedical Optics 22(7), 076011 (18 July 2017). https://doi.org/10.1117/1.JBO.22.7.076011

Received: 21 April 2017; Accepted: 23 June 2017; Published: 18 July 2017

ACCESS THE FULL ARTICLE

JOURNAL ARTICLE
10 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY