18 July 2017 Quantifying three-dimensional rodent retina vascular development using optical tissue clearing and light-sheet microscopy
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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.
© 2017 Society of Photo-Optical Instrumentation Engineers (SPIE)
Jasmine N. Singh, Jasmine N. Singh, Taylor M. Nowlin, Taylor M. Nowlin, Gregory J. Seedorf, Gregory J. Seedorf, Steven H. Abman, Steven H. Abman, Douglas P. Shepherd, 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 . Submission: Received: 21 April 2017; Accepted: 23 June 2017
Received: 21 April 2017; Accepted: 23 June 2017; Published: 18 July 2017
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