Human dentin, as an important calcified tissue in the body, plays significant roles in withstanding masticatory forces and has a complex hierarchical organization. Understanding the composition and ultrastructure of dentin is critical for elucidating mechanisms of biomineralization under healthy and pathological states. Here, atomic force microscope infrared spectroscopy (AFM-IR) and AFM-based amplitude modulation-frequency modulation (AM-FM) techniques were utilized to detect the heterogeneity in chemical composition and mechanical properties between peritubular and intertubular dentin at the nanoscale. AFM-IR spectra collected from peritubular and intertubular dentin contained similar vibrational bands in the amide regions (I, II and III), suggesting that collagen may exist in both structures. | Nanoscale chemical and mechanical heterogeneity of human dentin characterized by AFM-IR and bimodal AFM