In this study, the two methods of stiffness measurement(Spring constant) of cantilever were proposed for quantitative measurement in Atomic Force Microscopy(AFM). As the 1st method for the measurement of stiffness, the probe method, which is used in the measurement of the semiconductor mechanical and electrical properties, was applied to the measurement of the cantilever. Experiments by the probe method were performed finding the resistance value of cantilever. As the results, the resistance was measured differently along with the dimension and the thickness of cantilever that determined the stiffness(spring constant) of the lever. As the 2nd method, the vibration characteristics(Dunkerley expression) is used to obtain the stiffness of the complex structure which is combined by AFM cantilever and the standard cantilever. We measured the resonant frequency from the complex structure using the micro stages and stereo microscope. As the results, we confirmed that the vibration characteristics(Dunkerley expression) is effected the micro complex structure of AFM cantilever.