Quantifying three-dimensional rodent retina vascular development using optical tissue clearing and light-sheet microscopy.
J Biomed Opt
; 22(7): 76011, 2017 07 01.
Article
em En
| MEDLINE
| ID: mdl-28717817
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.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Retina
/
Vasos Retinianos
/
Microscopia de Fluorescência
Limite:
Animals
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article