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K.L. Njoh, M.E. Boulton, M. Rozanowska; Correlation of RPE Lipofuscin Autofluorescence Intensity with Drusen in AMD . Invest. Ophthalmol. Vis. Sci. 2003;44(13):1701.
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Purpose: To determine the lipofuscin autofluorescent (AF) intensity within retinal pigment epithelium (RPE) cells associated with drusen in early and late stage age-related macular degeneration (AMD). Methods: Four eyes (56-92 years) with age-related maculopathy (ARM) and five eyes with AMD (75-92 years) were fixed in formalin. Digital images of deparaffinized, unstained 5µm sections from the macula region and a region temporal and adjacent to the macular region were prepared at 450nm excitation and 490nm emission. Fluorescence intensity was measured, using an image analysis system, relative to a fluorescent standard. Between 2 and 13 drusen, per donor, were measured for lipofuscin AF intensity of the RPE cells overlying and immediately adjacent to the druse. Measurements of non-drusen associated RPE fluorescence (background) was made from sections containing no drusen. Results: Two main profiles of drusen associated RPE lipofuscin AF were observed, Type I and II. Of the 61 drusen analysed, 50 displayed Type I profile in which lipofuscin AF intensity in cells overlying the drusen was >16% below background, while fluorescent intensity was highest (>26% above background) in an annulus of RPE cells surrounding each drusen. The remaining 11 drusen displayed a Type II profile in which lipofuscin AF intensity in the cells overlying the drusen was >30% above background, while fluorescent intensity was lowest (>25% below background) within RPE cells surrounding each drusen. Cells overlying Type I drusen from AMD donors were significantly less fluorescent (p<0.05) than for ARM donors, while cells overlying Type II drusen of AMD donors were significantly more fluorescent (p<0.05) than those in ARM donors. Conclusions: The specific lipofuscin AF profiles associated with drusen are more pronounced in AMD donors. The profiles may represent altered RPE function in cells surrounding and overlying drusen.
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