Iontophoresis of horseradish peroxidase was used to label single auditory nerve fibers after determination of threshold tuning curves and rates of spontaneous discharge. The relation between characteristic frequency (CF) and cochlear longitudinal location is reconstructed from 52 labeled neurons in 16 cochleas. The length of the organ of Corti allotted to an octave of stimulus frequency increases steadily from low to high frequencies. Thus there is not a simple linear-distance-to-log frequency conversion. When comparing cochleas of different total length, the best predictor of CF at a given location is the distance from base or apex expressed as a percentage of the total length. The cochlear frequency map derived from these single-neuron labeling experiments is compared to maps derived by a number of different physiological and psychophysical techniques, and the significance of the similarities and differences is discussed.