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Coherent anti-Stokes Raman microscopy and spectroscopy for biological and biomedical sciences

Xi Huang, University of Nebraska - Lincoln


Many major discoveries in materials science, cellular biology, and other bio-fields require advanced visualization methods, for example microscopic techniques. Current microscopic techniques, such as traditional optical microscopy, fluorescence microscopy, and Raman spectroscopy, have been extensively applied in biological and biomedical sciences. They do play an important role for many applications. However, they have their own issues and problems. Traditional microscopy techniques cannot provide information related to chemical specificity. Fluorescence-based microscopy is powerful in many cases, but exogenous labels used often distort the interests and limit applications in specific labs targeting at tissue biopsies. Spontaneous Raman spectroscopy also suffers weak scattering efficiency problem.^ New microscopic technique is desired to address these problems. As a label-free technique, coherent anti-Stokes Raman scattering (CARS) provides intrinsic molecular vibrational information for chemical selectivity. This feature enables in vivo observation and detection towards clinical applications. On the other hand, the high-order dependence of CARS makes its signals fiver order stronger than spontaneous Raman scattering. Therefore, the objective of the research in this dissertation is 1) to introduce and demonstrate CARS microscopy and spectroscopy on biological and biomedical applications, 2) to improve imaging contrast of CARS microscopy, and 3) to develop a CARS system towards clinical applications.^ The work in this dissertation includes: 1) a CARS and multifunctional imaging system was developed and several multifunctional imaging applications were demonstrated, including microalgae cells, porcine carotid arterial walls, individual mouse liver cells and tissues, human muscle tissues, mouse breast cancer tissues, and mouse brain tissues; 2) CARS imaging contrast was enhanced by silica microspheres and FDTD simulation was conducted to support the experimental results; 3) Degeneration levels of human muscles with peripheral artery disease were discriminated by both spontaneous Raman (overall accuracy is 85.6%) and CARS spectroscopy (overall accuracy is 78.7%); 4) Breast tumor discrimination from normal tissues was conducted by CARS spectroscopy in conjunction with PC-DFA method and the overall accuracies are 98% and 95% for cross-validation and external-validation methods, respectively; and 5) a flexible delivery system for CARS was developed and built towards clinical applications. Through the work in this dissertation, CARS microscopy and spectroscopy shows a significant role on biological and biomedical sciences, and is a potential candidate towards clinical applications.^

Subject Area

Biomedical engineering|Electrical engineering

Recommended Citation

Huang, Xi, "Coherent anti-Stokes Raman microscopy and spectroscopy for biological and biomedical sciences" (2016). ETD collection for University of Nebraska - Lincoln. AAI10247651.