Abstract
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.
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References
Dicko C, Knight D, Kenney J, Vollrath F (2004) Structural conformation of spidroin in solution: a synchrotron radiation circular dichroism study. Biomacromolecules 5:758–767
Li G, Shao Z, Xie X, Chen X, Wang H, Chunyu L, Yu T (2001) The natural silk spinning process. Eur J Biochem 268:6600–6606
Shao Z, Vollrath F, Sirichaisit J, Young RJ (1999) Analysis of spider silk in native and supercontracted states using Raman spectroscopy. Polymer 40:2493–2500
Monti P, Taddei P, Freddi G, Asakura T, Tsukada M (2001) Raman spectroscopic characterization of Bombyx mori silk fibroin: Raman spectrum of Silk I. J Raman Spectrosc 32:103–107
Rousseau M-E, Lefèvre T, Beaulieu L, Asakura T, Pézolet M (2004) Study of protein conformation and orientation in silkworm and spider silk fibers using Raman microspectroscopy. Biomacromolecules 5:2247–2257
Fang G, Huang Y, Tang Y, Qi Z, Yao J, Shao Z, Chen X (2016) Insights into silk formation process: correlation of mechanical properties and structural evolution during artificial spinning of silk fibers. ACS Biomater Sci Eng 2:1992–2000
Ling S, Qi Z, Knight DP, Shao Z, Chen X (2011) Synchrotron FTIR microspectroscopy of single natural silk fibers. Biomacromolecules 12:3344–3349
Ling S, Qi Z, Shao Z, Chen X (2015) Determination of phase behaviour in all protein blend materials with multivariate FTIR imaging technique. J Mater Chem B 3:834–839. https://doi.org/10.1039/C4TB01808G
Ling S, Qi Z, Watts B, Shao Z, Chen X (2014) Structural determination of protein-based polymer blends with a promising tool: combination of FTIR and STXM spectroscopic imaging. Phys Chem Chem Phys 16:7741–7748
Ling S, Qi Z, Knight D, Shao Z, Chen X (2013) FTIR imaging, a useful method for studying the compatibility of silk fibroin-based polymer blends. Polym Chem 4:5401–5406. https://doi.org/10.1039/C3PY00508A
Kazarian S, Chan KLA (2010) Micro and macro-attenuated total reflection fourier transform infrared spectroscopic imaging. Appl Spectrosc 64:135–152
Chan KLA, Kazarian S (2003) New opportunities in micro- and macro-attenuated total reflection infrared spectroscopic imaging: spatial resolution and sampling versatility. Appl Spectrosc 57:381–389
Zhong J, Liu Y, Ren J, Tang Y, Qi Z, Zhou X, Chen X, Shao Z, Chen M, Kaplan DL, Ling S (2019) Understanding secondary structures of silk materials via micro- and nano-infrared spectroscopies. ACS Biomater Sci Eng 5:3161–3183
Baker M, Trevisan J, Bassan P, Bhargava R, Butler H, Dorling K, Fielden P, Fogarty S, Fullwood N, Heys K, Hughes C, Lasch P, Martin-Hirsch P, Effiong B, Sockalingum G, Sulé-Suso J, Strong R, Walsh M, Wood B, Martin F (2014) Using Fourier transform IR spectroscopy to analyze biological materials. Nat Protoc 9:1771–1791
Bassan P, Byrne HJ, Bonnier F, Lee J, Dumas P, Gardner P (2009) Resonant Mie scattering in infrared spectroscopy of biological materials – understanding the ‘dispersion artefact’. Analyst 134:1586–1593
Bassan P, Kohler A, Martens H, Lee J, Byrne H, Dumas P, Gazi E, Brown M, Clarke N (2010) Resonant Mie Scattering (RMieS) correction of infrared spectra from highly scattering biological samples. Analyst 135:268–277
Bassan P, Kohler A, Martens H, Lee J, Jackson E, Lockyer N, Dumas P, Brown M, Clarke N (2010) RMieS-EMSC correction for infrared spectra of biological cells: extension using full Mie theory and GPU computing. J Biophotonics 3:609–620
Ling S, Qi Z, Knight DP, Huang Y, Huang L, Zhou H, Shao Z, Chen X (2013) Insight into the structure of single Antheraea pernyi silkworm fibers using synchrotron FTIR microspectroscopy. Biomacromolecules 14:1885–1892
Lasch P, Haensch W, Naumann D, Diem M (2004) Imaging of colorectal adenocarcinoma using FT-IR microspectroscopy and cluster analysis. Biochim Biophys Acta 1688:176–186
Lasch P, Diem M, Hänsch W, Naumann D (2006) Artificial neural networks as supervised techniques for FT-IR microspectroscopic imaging. J Chemom 20:209–220
Acknowledgements
We thank the BL01B beamline of National Center for Protein Science Shanghai (NCPSS) at Shanghai Synchrotron Radiation Facility.
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Zhong, J., Zhou, X., Ye, C., Yu, W., Tang, Y. (2021). Using FTIR Imaging to Investigate Silk Fibroin-Based Materials. In: Ling, S. (eds) Fibrous Proteins. Methods in Molecular Biology, vol 2347. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1574-4_18
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DOI: https://doi.org/10.1007/978-1-0716-1574-4_18
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