An understanding of whole-cell mechanical behavior can provide insight into cellular responses to mechanical loading and diseases in which such responses are altered. However, this aspect of cellular mechanical behavior has received limited attention. In this study, we used the atomic force microscope (AFM) in conjunction with several mechanical characterization methods (Hertz contact theory, an exponential equation, and a parallel-spring recruitment model) to establish a mechanically rigorous method for measuring and characterizing whole-cell mechanical behavior in the deformation range 0-500 nm. Using MC3T3-E1 osteoblasts, measurement repeatability was assessed by performing multiple loading cycles on individual cells. Despite variability in measurements, repeatability of the measurement technique was statistically confirmed. The measurement technique also proved acceptable since only 5% of the total variance across all measurements was due to variations within measurements for a single cell. The parallel-spring recruitment model, a single-parameter model, accurately described the measured nonlinear force-deformation response (R2 > 0.99) while providing a mechanistic explanation of whole-cell mechanical behavior. Taken together, the results should improve the capabilities of the AFM to probe whole-cell mechanical behavior. In addition, the success of the parallel-spring recruitment model provides insight into the micromechanical basis of whole-cell behavior.