應用相異波長偏振二倍頻顯微術於膠原蛋白與肌肉纖維

Wavelength Dependence of Polarization Second Harmonic Generation Microscopy of Collagen and Muscle Fiber

黃君平(Chun-Ping Huang)

指導教授 ： 董成淵

國立臺灣大學/理學院/物理研究所/碩士(2013年)

https://doi.org/10.6342/NTU.2013.00614

摘要

近年來，偏振二倍頻顯維術逐漸受到重視，利用不同的線偏振光產生二倍頻訊號來分析材料結構，成為非線性光學上的重要研究方法。而材料的二階極化率與其結構密切相關，研究不同材料的二階極化率可用來增加彼此的影像對比程度。
實驗上我們旋轉半波片與四分之一波片，取得線偏振光，因為分光鏡也會影響雷射光的偏振特性，造成雷射功率改變，實驗前先測量螢光劑的螢光強度當成校正功率的工具。
針對大鼠尾巴肌腱、大鼠氣管軟管、大鼠腿部肌肉，其分子理論模型都是圓柱對稱，二階極化率只有三個獨立變數，且上述樣品的自然共振波長約在200nm∼300nm左右，實驗的雷射波長在700nm∼900nm，故我們引入了Kleinman Symmetry條件，當材料的自然共振頻率遠高於入射光頻率時，其二階極化率張量不隨入射光頻率而改變。本論文從廣義的非中心對稱材料的位能出發，得到電子在三維系統的自然共振頻率、入射光頻率及二階極化率張量的關係，並測量上述三種材料在不同波長下的二階極化率比值趨近一定值，驗證了在700nm∼900nm的波長範圍下，這三種生物材料使用圓柱對稱及Kleinman Symmetry假設條件的理論模型值得相信。

關鍵字

偏振二倍頻 ； Kleinman Symmetry ； 膠原蛋白 ； 肌肉纖維 ； 二階極化率

並列摘要

Recently, the polarization-resolved Second Harmonic Generation (SHG) microscopy has gained much attention from researchers in the field of non-linear optics and becoming an important researching method. The second order susceptibility(χ(2)) is closely related to material structure. The χ(2) in different materials can used to enhance the contrast of SHG images.
The linear polarization light is usually be done by rotating the half-wave plate and quarter-wave plate. However, the dichroic mirror also alters the power of laser. Therefore, in this study, we measured the two-photon fluorescence of a fluorescent agent(0.5mg/ml,sulforhodamine b,s1420-5g) to calibrate the change of SHG signal.
The structure of rat-tail tendons(RTT), rat-trachea cartilages(RTC), and rat-leg muscles(RLM) are based on the cylindrical symmetry theory. There have only three independent variables of χ(2). The natural resonance frequency of RTT,RTC,RLM are about 200nm ~ 300nm, and the excitation laser wavelength is about 700nm ~ 900nm. The Kleinman Symmetry assumption was adopted in this study to hold χ(2) as a constant. By introducing the general potential of non-centrosymmetric material, it was found that the χ(2) ratio of RTT, RTC, and RLC with different wavelengths are approaching constants in the three dimension system.These results suggested that applying the hypothesis of the cylindrical model and Kleinman Symmetry to such materials is suitable.

並列關鍵字

Second Harmonic Generation ； Kleinman Symmetry ； Collagen ； Muscle Fiber ； Second Order Susceptibility

參考文獻

[6] J. Zhang, R. Chen, W. Liu, Z. Chen, L. Shu, Y. Liu, "The spectra character of photodegraded the pyridinoline cross-links by Hypocrellin B" The 9th International Conference on Photonics and Imaging in Biology and Medicine, Journal of Physics: Conference Series 277,012016 (2011)

[1] Paul J Campagnola and Leslie M Loew " Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms " Nature Biotechnology 21, 1356 - 1360 (2003)

[2] W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, W. W. Webb "Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation " Jun 10;100(12):7075-7080.(2003)

[3] Shoulders MD, Raines RT. "Collagen structure and stability". Annu Rev Biochem

78:929-58.( 2009)