受激拉曼散射（SRS）顯微術是一種非常有價值的技術，它可以觀測特定的化學鍵，而無需使用任何標記，然而與螢光相比SRS的靈敏度較低，主要是由於其較低的拉曼截面。為了克服這一限制，人們提出了電子預共振（EPR）效應，通過電子共振來增強核振動從而提高拉曼截面，這種方法涉及調整激發波長，以緊密匹配目標樣品的吸收峰。為了證明EPR-SRS的有效性，需要同時調整pump和Stokes光束的波長，以使它們與目標樣品的吸收峰相吻合，同時保持拉曼位移（Ω）不變。 在這項研究中，我們使用具有極寬頻（600-1300奈米）的多重薄片展頻光源（MPC）來產生pump和Stokes光束，從而實現了“雙波長可調”的特性。我們利用Alexa 635螢光染料的C=C鍵結來展示整個EPR範圍的SRS訊號增強效果，而我們的實驗結果與Albrecht A-term pre-resonance模型非常吻合，顯示出SRS信號強度顯著增加了150倍。此外我們展示了 EPR-SRS 生物成像在 Alexa 635 染色之果蠅腦樣本上的應用，證實了 MPC-EPR-SRS 在生物成像中的可行性和潛力。

Stimulated Raman scattering (SRS) microscopy is a valuable technique for capturing specific chemical bonds without the need for labeling. However, SRS has lower sensitivity compared to fluorescence, mainly due to the challenge of low Raman cross-section. To overcome this limitation, electronic pre-resonance (EPR) effects, which leverage electronic resonance to enhance nuclear vibrations and thereby the Raman cross-section, are well-established. This approach involves tuning the excitation wavelength to closely match the absorption peak of the target sample. In order to demonstrate the effectiveness of EPR-SRS, it is necessary to simultaneously adjust the wavelengths of the pump and Stokes beams to align with the absorption peak of the target sample while maintaining the Raman shift (Ω).
In this study, we employ a multiple-plate continuum (MPC) light source with an extremely broad bandwidth (600-1300 nm) to generate both pump and Stokes beams, providing "dual-wavelength tunability." We showcase the enhancement of the entire EPR range using the C=C mode of the Alexa 635 fluorescent dye. Our experimental findings align closely with the Albrecht A-term pre-resonance model, revealing a remarkable 150-fold increase in SRS signal intensity. Furthermore, we demonstrate the application of EPR-SRS bioimaging on Drosophila brain samples stained with Alexa 635, affirming the feasibility and potential of MPC-EPR-SRS in bioimaging.

摘要.................................................................1
Abstract.............................................................2
Acknowledgements.....................................................4
Table of Contents....................................................5
List of Figures......................................................7
Chapter 1. Introduction.............................................11
Chapter 2. Theory...................................................14
Chapter 3. Materials and Methods....................................28
Chapter 4. Experimental Results.....................................42
Chapter 5. Discussion and Conclusion................................52
References..........................................................55

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