光纤激光生物传感技术及其肿瘤标志物检测应用研究

肿瘤标志物的有效检测对于肿瘤早期筛查与诊断、治疗方案指导以及治疗效果与预后评估都具有不可或缺的作用。在光学传感器领域，光纤生物传感器在肿瘤标志物检测方面的应用已成为重要的研究方向。鉴于肿瘤标志物在样本中的稀缺性及其所处环境的复杂性，对于传感器的灵敏度、解调分辨率及特异性都提出了更高的要求。而光纤激光，以其独特的窄线宽和高信噪比特性，为光纤检测技术光谱分辨能力的提升开辟了新路径。本论文深入挖掘了光纤激光的优势与潜能，提出了光纤激光生物传感器的一系列设计构想，旨在实现对肿瘤标志物的精准特异性检测。主要研究内容如下：

（1）提升光谱分辨能力方面：提出一种基于光纤套索结构的光纤激光生物传感方案，利用激光信号解调干涉光谱信息，结合高分辨率光谱分析，将传感器的光谱分辨能力（相比于传统干涉光谱分析）提升了4个数量级，实现对人血清中肿瘤标志物CEACAM5的高精度检测。通过理论分析套索结构的模式干涉、谐振腔中的模式传输特性，结合公式推导、RSOFT光波导模式传输仿真和实验验证，实现了激光波长与套索结构干涉光谱信息的精确对应。在对肿瘤标志物CEACAM5的特异性检测中，传感器的检测下限为9.6 ng/mL，与文献中灵敏度高出10倍的生物传感器性能相当，证明了激光检测技术在高精度生物检测方面的优势。

（2）增强传感器响应灵敏度方面：提出一种磁珠增强的微纳套索结构光纤激光传感方案，通过结构设计增敏与磁珠信号放大的共同作用，实现对人血清中肿瘤标志物CEACAM5的超高灵敏检测。通过利用多物理场软件COMSOL仿真、理论分析和实验验证，证实了所引入的花生级联结构、微纳拉锥和弯曲套索结构设计对增强灵敏度的有效性。同时，采用“抗体-抗原-抗体-磁珠”夹心结构，进一步放大了信号响应，实现了对肿瘤标志物CEACAM5的超灵敏检测，检测下限低至0.11 ng/mL，远低于医学筛查阈值（5 ng/mL）。与医学临床数据对比，对人血清样本中标志物含量的检测结果偏差在1.9%~9.8%之间，展现出良好的临床应用潜力。

（3）提升检测分辨率方面：借鉴交叉学科思路，提出一种基于微波光子解调的双波长激光生物检测方案，将光谱波长变化转换为频谱频率变化，将检测分辨率提升了2个数量级。通过理论研究和公式推导，分析了双波长激光的波长差值与频域信号的对应关系，并优化了系统的频谱解调灵敏度响应。实验结果表明：该方案的折射率检测分辨率达到5.26◊10^-8 RIU，对肿瘤标志物CEACAM5的特异性检测下限低至0.076 ng/mL，显著优于传统的光谱波长分析方法。

（4）低成本化的高分辨率检测：设计并构建了微纳拉锥D型光纤激光器，深入研究激光腔内的多纵模拍频与偏振模拍频信号，用低成本的电域解调方案实现高分辨率频域检测。结合多物理场软件COMSOL仿真，对其双折射特性、灵敏度响应机制进行了理论分析与实验验证。实验结果显示，该系统的折射率检测分辨率为7.40×10^-6 RIU，优于已报道的基于光谱波长分析或散斑分析的众多光纤传感器。该方法在生化分析、肿瘤标志物检测等领域具有巨大的应用潜力。用3D打印模具和PDMS制作了微流控芯片，对光纤传感器做集成封装，进一步增强了传感器性能的稳定性。

综上所述，本论文针对肿瘤标志物检测的难点，结合光纤激光技术的优势，对光纤生物传感器的检测性能进行了系统的提升研究。具体而言，实验验证了基于激光信号分析比传统干涉光谱分析的光谱分辨能力（FOM参量）4 个数量级的提升，传感器响应灵敏度的增强，以及检测分辨率2个数量级的提高。还探索了低成本、高分辨率的频域解调方法，使光纤激光生物传感技术在肿瘤标志物检测方面更具实用性和可行性。

The detection of tumor markers plays a significant role in improving the early screening and diagnosis rate of tumors, guiding the selection of treatment options, and assessing treatment effectiveness and prognosis. The research on optical fiber biosensors and their applications in tumor marker detection represents a major branch in the field of optical biosensors. The low abundance of tumor markers in test samples and the complexity of the sample environment pose higher demands on the sensitivity, demodulation resolution, and specificity of sensors. The narrow linewidth and high signal-to-noise ratio of fiber lasers provide a potential pathway to enhance the spectral resolution of fiber-optic detection techniques. This dissertation leverages the advantages and characteristics of fiber lasers to propose a fiber laser biosensor for the specific detection of tumor markers. The main research contents are as follows:

(1) Enhancing spectral resolution: A lasso shaped fiber laser biosensor is proposed. By demodulating interference spectral information using laser signals, the spectral resolution of the sensor is improved by four orders of magnitude (compared to traditional interference spectrum analysis), enabling high-precision detection of the tumor marker CEACAM5 in human serum. Through theoretical analysis of mode interference in the lasso structure and mode transmission characteristics in the resonant cavity, combined with formula derivation, RSOFT optical waveguide mode transmission simulation, and experimental validation, precise correspondence between laser wavelength and interference spectral information of the lasso structure is achieved. In specific detection of CEACAM5, the detection limit of the sensor is 9.6 ng/mL, which is comparable to the performance of biosensors with 10 times higher sensitivity reported in the literature, demonstrating the advantage of laser detection technology in high-precision biological detection.

(2) Enhancing sensor response sensitivity: A magnetic-beads-enhanced peanut structure cascaded lasso shaped fiber laser biosensing scheme is proposed. Through the combined effect of structural design sensitization and signal amplification of magnetic bead, achieving ultra-sensitive detection of human serum tumor marker CEACAM5. By utilizing physics simulation of software COMSOL, theoretical analysis, and experimental validation, the effectiveness of structural innovation design to enhance sensitivity (introduced peanut cascade structure, tapered microfiber, and bent lasso design) is confirmed. Meanwhile, an "antibody-antigen-antibody-magnetic bead" sandwich structure is employed, further amplifying the signal response, and enabling ultrasensitive detection of the tumor marker CEACAM5 with a detection limit as low as 0.11 ng/mL, which is significantly lower than the medical screening threshold (5 ng/mL). Compared with clinical data, the deviation of the detection results for marker content in human serum samples is between 1.9% and 9.8%, demonstrating good clinical application potential.

(3) Improving detection resolution: A dual-wavelength laser biosensing scheme based on microwave photonic demodulation technology is proposed, which converts spectral wavelength changes into radio frequency spectrum variations， and representing a two-order-of-magnitude improvement compared to spectral wavelength demodulation resolution. Through theoretical research and formula derivation, the correspondence between the wavelength difference of the dual-wavelength laser and the frequency domain signal is analyzed, and the sensitivity response is optimized. Experimental results demonstrate that the refractive index detection resolution of this scheme is as high as 5.26×10^-8 RIU. When specifically detecting the tumor marker CEACAM5, the detection limit of this scheme is as low as 0.076 ng/mL, significantly superior to traditional spectral wavelength analysis methods.

(4) Low-cost high-resolution detection: A tapered micro-D-shaped fiber laser was designed and constructed, and an in-depth study of multi-longitudinal mode beating and polarization mode beating signals within the laser cavity was conducted. High-resolution frequency domain detection was achieved using low-cost electrical methods. Combined with physics simulation of software COMSOL, theoretical analysis and experimental validation were performed on its birefringence characteristics and sensitivity response mechanisms. Experimental results showed that the refractive index detection resolution of the system is 7.40×10^-6 RIU, which is superior to many reported optical fiber sensors based on spectral wavelength analysis or speckle analysis. This method has significant application potential in fields such as biochemical analysis and tumor marker detection. A microfluidic chip was fabricated using 3D printed molds and PDMS to integrate and encapsulate the fiber optic sensor, further enhancing the stability of sensor performance.

In summary, this paper systematically enhances the sensing performance of fiber optic biosensors by addressing the challenges of tumor marker detection and leveraging the advantages of fiber laser technology. Specifically, the experimental results verify that the spectral resolution (FOM parameter) of laser-based signal analysis is improved by 4 orders of magnitude, the sensor response sensitivity is enhanced, and a two-order-of-magnitude increase in detection resolution. Additionally, low-cost, high-resolution frequency domain demodulation method has been explored, making fiber laser biosensing technology more practical and feasible for tumor marker detection.

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