Fluorescence/Raman intensity ratio for monitoring the pathologic state of human lens

The authors have put quantitation of human lens fluorescence on a rational basis by using the accompanying Raman signal from lens protein as a normalization factor. The intensity ratio, Fluorescence/Raman (F/R), may be used to compare lenses of different ages when the exciting wavelength is long enough to give a measurable Raman signal. In younger lenses excited at 457.9 or 514.5 nm, the F/R shows a log increase with age. Older lenses, above 60 years of age, excited at 647.1 nm give a steeply rising sigmoid curve. In developing this procedure, the authors found that for each lens there is a characteristic wavelength that is called the critical wavelength (lambda critical). At wavelengths longer than lambda critical the Raman signal appears in the absence of a broad fluorescence peak; at shorter wavelengths the fluorescence intensity increases enough to overwhelm the Raman signal. For normal lenses, clear and not heavily pigmented, the lambda critical is age dependent, giving a curve that is a flattened sigmoid approximating a straight line.