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M. Friedlander, L. Scheppke, E. Aguilar, E. Banin, M. Ritter; In vivo imaging of mouse ocular vascular development using confocal microscopy. . Invest. Ophthalmol. Vis. Sci. 2004;45(13):2199.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: We have developed an in vivo imaging method for visualization of mouse ocular vasculature. This method was used to study choroidal, retinal, hyaloidal and iris vascular development from birth to young adulthood. Methods: Multiphoton scanning laser confocal microscopic imaging techniques were used. Anesthetized mice are mounted on an inverted microscope using a custom–modified stage. Vessels were visualized using intravenously–injected fluorescent–labeled dextrans or Tie–2 Green Fluorescent Protein transgenic mice. Confocal imaging was carried out on a Bio–Rad Radiance 2100 MP system equipped with a Spectra–Physics Tsunami Ti:sapphire laser. Results: With this imaging approach, we generated 3–dimensional reconstructions of the mouse ocular vasculature. High resolution images of the central retina, hyaloid, tunica vasculosa lentis, and iris were collected. These images were obtained from living animals under anesthesia which allows for serial imaging of the same eye for an indefinite period. Phototoxicity was minimized with the use of pulsed, low energy infrared light. In enucleated eyes, we have also obtained high resolution images of the choroidal vasculature. The vascular developmental pattern observed using this novel in vivo imaging technique is generally in accord with that generated from serial analysis of ocular tissue specimens obtained from fixed, post–mortem eyes. In addition, it allows enhanced visualization of the hyaloid system and permits temporal, as well as spatial, relationships to be observed in live tissue. Conclusions: In vivo confocal imaging permits the study of ocular vasculature development in living animals, providing new insights into the dynamics of vascular development in the eye.
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