New-generation biomaterials are information-rich and incorporate biologically active components derived from Nature. The chapter reviews current approaches for the realization of biomimetic coatings for tissue engineering applications, via functionalization with bioactive molecules such as native long chains of extracellular matrix (ECM) proteins as well as short peptide sequences derived from intact ECM proteins. Biomimetic materials provide cues for cell-matrix interactions, favouring cell adhesion, proliferation, spreading, and differentiation. The recently developed scanning probe techniques for optical and spectroscopic characterization of surfaces are also described, providing advanced topographical imaging and sensitive force measurements. The spectroscopic imaging of surfaces at the nanoscale provides insight into the function and structure of biomimetic molecules and represents a tool for tailoring the surface design.