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A.D. Marmorstein, A.M. Floyd; Fundus Autofluorescence Imaging in Post–Mortem, Intact Human and Pig Eyes . Invest. Ophthalmol. Vis. Sci. 2005;46(13):4286.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To test the hypothesis that fundus fluorescence ratiometry might be a useful diagnostic tool for retinal disease by distinguishing autofluorescence associated with lipofuscin from autofluorescence associated with sub–RPE deposits. Methods: Human and pig eyes were imaged using a modified fundus camera after scraping of the corneal epithelium and phacoemulsification of the lens and lens capsule or after dissection of the anterior segments and vitreous. Reflected light images were captured and used as an index. Fluorescence was excited using using 405,nm, 430nm, 450nm, or 470nm light. Emissions bandwidth was varied using different combinations of barrier filters. Two emissions bandwidths were simultaneously collected. A ratiometric map of the fundus was then created using Metamorph software. After imaging the eyes were fixed and processed for paraffin embedding and stained using hematoxylin and eosin. Results: We have successfully generated both reflected light and fluorescence images from human and pig eyes. Initial data suggest that significant differences in Ratiometric comparisons indicate significant differences in emissions from the wavelengths used that may correlate with the development of sub–RPE deposits. Conclusions: This study provides the basis for continuing studies to determine whether features identified using fundus fluorescence ratiometry correspond to pathologic deposits associated with diseases such as age–related macular degeneration.
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