Biomaterials characterization is a crucial first step in the biomaterial evaluation process towards practical applications. The characterization procedure is designed to evaluate many different properties of biomaterials aimed for utilization in medical devices. The extent to which a biomaterial needs to be characterized depends upon the biomaterial type, the end use of the device, and the function of the biomaterial within the device. Different advanced methods for biomaterials characterization will be discussed: physico-chemical methods (e.g. Fourier transform infrared (FTIR) spectroscopy, Raman spectroscopy, nuclear magnetic resonance (NMR) spectroscopy, X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD) analysis, field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), energy-dispersive X-ray spectroscopy (EDXS), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), differential thermal analysis (DTA)), mechanical testing methods (e.g. tensile and impact strength, elastic modulus, hardness, tension, compression, shear and flexure) and biological testing (e.g. biocompatibility, cytotoxicity, antibacterial activity, immunomodulation, blood-biomaterial interactions, functionality).