In this study, mechanical properties of guinea pig outer hair cells (OHCs) were measured by atomic force microscopy (AFM). First, in order to confirm the availability of AFM for measurement of the mechanical properties of the OHC, Young's moduli of the OHCs measured in this study were converted into stiffnesses using a one-dimensional model of the cell and then compared with the values reported in the literature. Next, the difference in local mechanical properties of the OHC along the cell axis was measured. Finally, the relationship between Young's modulus in the middle region of the OHC and the cell length was evaluated. The results were as follows. (1) AFM is an adequate tool for the measurement of mechanical properties of the OHC. (2) Mechanical properties in the apical region of the OHC are a maximum of three times larger than those in the basal and middle regions of the cell. (3) Young's modulus in the middle region of a long OHC obtained from the apical turn of the cochlea and that of a short OHC obtained from the basal turn or the second turn are 2.0+/-0.81 kPa (n=10) and 3.7+/-0.96 kPa (n=10), respectively. In addition, it was found that Young's modulus decreases with an increase in the cell length.