The distribution of anthocyanins in grape (Vitis vinifera L.) bunches from the Sangiovese cultivar was measured nondestructively by chlorophyll fluorescence imaging using two excitation light bands at 550 and 650 nm in sequence. The pixel intensity in the derived logarithm of the fluorescence excitation ratio image was directly related, by an exponential function (r2 = 0.93), to the anthocyanin concentration of berry extracts. The method will be useful for the assessment of the heterogeneity of anthocyanin accumulation in berries that is known to depend on physiologic and climatic factors. It can also represent a new, rapid and noninvasive technique for the assessment of grape ripening and the appropriate time of harvest.