Label-Free Characterization of Collagen Crosslinking in Bone-Engineered Materials Using Nonlinear Optical Microscopy

Chao Wei Hung, Nirmal Mazumder, Dan Jae Lin, Wei Liang Chen, Shih Ting Lin, Ming Che Chan, Guan Yu Zhuo

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)


Engineered biomaterials provide unique functions to overcome the bottlenecks seen in biomedicine. Hence, a technique for rapid and routine tests of collagen is required, in which the test items commonly include molecular weight, crosslinking degree, purity, and sterilization induced structural change. Among them, the crosslinking degree mainly influences collagen properties. In this study, second harmonic generation (SHG) and coherent anti-Stokes Raman scattering (CARS) microscopy are used in combination to explore the collagen structure at molecular and macromolecular scales. These measured parameters are applied for the classification and quantification among the different collagen scaffolds, which were verified by other conventional methods. It is demonstrated that the crosslinking status can be analyzed from SHG images and presented as the coherency of collagen organization that is correlated with the mechanical properties. Also, the comparative analyses of SHG signal and relative CARS signal of amide III band at 1,240 cm-1to CH2 band at 1,450 cm-1of these samples provide information regarding the variation of the molecular structure during a crosslinking process, thus serving as nonlinear optical signatures to indicate a successful crosslinking.

Original languageEnglish
Pages (from-to)1-11
Number of pages11
JournalMicroscopy and Microanalysis
Issue number3
Publication statusPublished - 06-2021

All Science Journal Classification (ASJC) codes

  • Instrumentation


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