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J. J. McGarvey, J. R. Beattie, A. M. Pawlak, M. E. Boulton, A. W. Stitt; Raman Spectroscopic Evaluation of Age Related Changes in Bruch’s Membrane. Invest. Ophthalmol. Vis. Sci. 2010;51(13):2778.
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© ARVO (1962-2015); The Authors (2016-present)
Bruch’s membrane (BM) is a highly dynamic matrix that is integral to RPE and outer retinal function. Reactive intermediates of metabolism are elevated during ageing and the adducts formed can modify the collagens of BM and this has a critical role in the development of AMD. Analysing advanced glycation / lipoxidation endproducts (collectively termed AGEs) normally requires destructive, highly specific analyses. The purpose of this study is to investigate the potential of the non-invasive technique of Raman spectroscopy to multiplex age-related biochemical information.
BM from human post-mortem eyes were flat mounted and probed using a Raman microscope. A spectral database of a wide range of collagens, fats, porphyrins and metabolic adducts was recorded. Principal component analysis was used to extract the spectral signals of constituents which were compared to database for ID.
The Raman data yielded 60 unique biochemical constituents including collagens, heme, cytochrome, fatty acid lipids, cholesterol, oxidised lipids and a number of AGE adducts (CML,CEL,CEP,GH-1,HHE,GO,DHP-lys and pentosidine). AGEs in particular showed significant increasing trends against chronological age, especially between 70 and 80 years of age (Fig 1). In addition to AGE changes with age, the Raman signal simultaneously demonstrated a rise in heme and cholesterol in BM
Raman is able to simultaneously detect a large number of biochemical changes with chronological age, including AGE adducts that have been implicated in age-related retinal pathology.
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