Using FTIR Imaging to Investigate Silk Fibroin-Based Materials

The secondary structures of silk fibroin (SF) are critical in the determination of the mechanical properties of the animal silks. Different characterization techniques, such as X-ray diffraction, nuclear magnetic resonance, Raman spectroscopy, and Fourier transform infrared (FTIR) technique, have been applied to study the secondary structure of animal silks. Among these techniques, FTIR is most widely used as it is sensitive to all secondary structures of proteins. Especially with the development of FTIR imaging, it is now possible to image the secondary structures of proteins at the micrometer scale, so as to understand the spatial distribution of proteins and the interaction of proteins with other materials at specific locations of interest. In this chapter, we present the methods and protocols of FTIR imaging to silk protein-based materials. We primarily introduce how to set up the instruments and accessories, as well as how to choose the appropriate imaging methods and sample preparation methods according to sample morphologies. The critical protocols for data analysis are also introduced in the last section.