The optical density of macular pigment was measured at twelve retinal locations in ten subjects by minimum motion photometry, comparing 460 nm with 580 nm or 550 nm. Fundus autofluorescence images were obtained for the same subjects with a scanning laser ophthalmoscope. Optical density was computed from mean calibrated grey-scale values for a central circular field and for annular segments, identical to areas tested psychophysically, and for complete annuli. Psychophysical assessments of optical density were similar irrespective of whether 550 nm or 580 nm was used. Optical density values derived psychophysically showed a linear correlation with assessments based on identical sampled areas of annular segments (slope = 0.98, r2 = 0.97) or complete annuli (slope = 0.89, r2 = 0.96) in autofluorescence images.