Advanced numerical simulations of the mechanical behavior of human skin require thorough calibration of the material's constitutive models based on experimental ex vivo mechanical tests along with images of tissue microstructure for a variety of biomedical applications. In this work, a total of 14 human healthy skin samples and 4 additional scarred skin samples were experimentally analyzed to gain deep insights into the biomechanics of human skin. In particular, second harmonic generation (SHG) microscopy was used to extract detailed images of the distribution of collagen fibers, which were subsequently processed using a three-dimensional Fourier transform-based method recently proposed by the authors to quantify the distribution of fiber orientations. Mechanical tests under both biaxial and uniaxial loading were performed to calibrate the relevant mechanical parameters of two widely used constitutive models of soft fiber-reinforced biological tissues that account for non-symmetrical fiber dispersion. The calibration of the models allowed us to identify correlations between the mechanical parameters of the constitutive models considered. STATEMENT OF SIGNIFICANCE: Constitutive models for soft collagenous tissues can accurately reproduce the complex nonlinear and anisotropic mechanical behavior of skin. However, a comprehensive analysis of both microstructural and mechanical parameters is still missing for human skin. In this study, these parameters are determined by combining biaxial mechanical tests and SHG stacks of collagen fibers on ex vivo healthy human skin samples. The constitutive parameters are provided for two widely used hyperelastic models and enable accurate characterization of skin mechanical behavior for advanced numerical simulations.