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M. C. Butler, J. M. Sullivan; A Novel Real-Time in-vivo Mouse Retinal Imaging System. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3103.
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To develop an efficient low cost instrument for robust illumination and imaging of the live mouse retina through dilated pupils with constant broad band or band-limited optical stimuli. Use this instrument to quantify the extent of subretinal vector delivery.
Optical constraints for imaging of the retina of the small eye during or after subretinal injections in preclinical gene therapy trials have not been systematically evaluated. A number of devices were reported to support successful imaging, but these have never been compared, and their design underpinnings are not well established. We evaluated the optical properties of a number of illumination systems used for retinal imaging in rodents. Gauging the limitations of these devices for our purposes, we designed a parafocal, paraxial illumination system for Greenough-type stereo dissecting microscopes. The optical launch design is optimized in that the selected lamp has an optical output matched to the numerical aperture of the fiberoptic delivery system, and the variable-focus optical system at the stereo microscope is optimized to the constraints of the small dilated pupil (~1mm). Filters are used to control spectral bandwidth. Field-of-view is constrained by the long working distance (needed for surgical manipulations), the small pupil aperture, and the high optical power of the mouse eye, but can be compensated by eye positioning in order to observe the entire retina. A color CCD camera is coupled to the microscope for image capture.
The retinal imaging system delivers an adjustable narrow beam to the dilated pupil to fill this aperture with minimal vignetting. The optic nerve, vasculature, and posterior pole are crisply visualized and the entire anterior edge of the retina can be observed through eye positioning. Normal and degenerative retinal phenotypes can be followed over time. Subretinal injection procedures are followed in real time.
a novel device is established for real time viewing and image capture of the retina during subretinal injections for preclinical gene therapy studies.
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