Diffuse spectral fundus reflectance measured using subretinally placed spectralon

The diffuse fundus reflectance and the spectral transmittance of the swine sensory retina was measured in vivo using intravitreal illumination. Pars plana vitrectomy and intravitreal manipulations were performed on a female American Yorkshire domestic swine. Light from a scanning monochromator was coupled into a fiber optic intraocular illuminator inserted into the vitreous. A 1.93-mm ² region of the illuminated fundus was imaged from an oblique illumination angle. Multispectral retinal images were acquired for four experimental conditions: the eye (1) prior to vitrectomy, (2) after vitrectomy, (3) after insertion of a Spectralon disk super-retinally, and (4) after subretinal insertion of the disk. The absorption of melanin and hemoglobin in the red wavelengths was used to convert relative spectral reflectance to absolute reflectance. The flux scattered from the super-retinal Spectralon was used to correct for scattering in the globe. The transmittance of the sensory retina was measured in vivo using the scatter corrected subretinal Spectralon disk reflectance. The hemoglobin and melanin components of the spectrum due to scattered light were removed from the retinal transmission spectrum. The in vivo spectral transmittance of the sensory retina in this swine was essentially flat across the visible spectrum, with an average transmittance >90%.