The structures and mechanical properties of human tissues are significantly influenced by water. The functionality of the human cornea can be linked to the hydrated collagen fibers. By applying quantitative dynamic atomic force microscopy to investigate morphological and mechanical property variations of corneal stroma under different hydration levels, we found that the collagen fibers in the stromal tissue show the specific periodicities and the stiffness of giga-Pa magnitude at 40% humidity. However, under increasing hydration, the collagen fibers clearly show nanoparticle structures along the fibers with the stiffness in mega-Pa magnitude. By increasing the hydration time, the stroma regains the fiber structure but with larger diameter. The age-dependency in stiffness was further investigated. The interplay of structures and nanomechanical mapping may be applied for the future diagnosis and assessment or even pathologic analysis.