Programm                 "Degeneration und Regeneration– Grundlagen, Diagnostik und Therapie"

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Visualization of Retinal Pigment Epithelial Cells in vivo Using Digital High Resolution Confocal Scanning Laser Ophthalmoscopy

Bindewald A., Jorzik J. J., Schütt F., Lösch A., Holz F. G.
Department of Ophthalmology, University of Heidelberg

Purpose: The retinal pigment epithelium (RPE) plays a central pathophysiological role in various retinal diseases. Conventional imaging methods do not allow resolution of the RPE on a single cell level in vivo. Our purpose is to describe the delineation and visualization of RPE cells in vivo using a new confocal scanning laser ophthalmoscope (cSLO) with enhanced resolution.
Method: Digital fundus autofluorescence (AF) imaging was performed with a cSLO (Heidelberg Retina Angiograph 2, Heidelberg Engineering) after excitation with an optically pumped solid state laser (488 nm) and detection of emitted light above 500 nm. Comparable in vitro images of human donor eyes were obtained after removal of the neurosensory retina with the cSLO and in flatmount preparations by fluorescence microscopy.
Results: In presence of clear optical media, sufficient age-related accumulation of RPE lipofuscin and peripheral to absorbing macular pigment the hexagonal RPE pattern was visualized and single cells identified. Intensity of autofluorescence varied considerably between single cells. The AF pattern was in accordance with in vitro images regarding size and shape after removal of overlying retina. Identification in the AF-imaging mode is based on apical and central distribution of absorbing melanin g

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