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目的:研究用于癌症诊断与治疗的光敏剂血卟啉(HPD)的超快光动力学过程;方法:采用超短脉冲激光光谱技术和皮秒时间相关单光子计数系统,测量经血卟啉培养的活体癌细胞与正常细胞的荧光光谱、荧光寿命特性及荧光峰值强度随时间变化曲线,并测量单一细胞内部不同位置的荧光寿命特性;结果:癌细胞样品在645nm处具有特有的光谱谱峰;癌细胞样品荧光寿命的快成分约150ps慢成分约1200ps,而正常细胞样品快成分约300ps慢成分约2500ps;癌细胞样品的荧光峰值强度经12小时衰减约10%,而正常细胞样品衰减约55%;在细胞内部荧光寿命300ps的快成分十分显著,且中心部位血卟啉浓度最高;结论:癌细胞与正常细胞的荧光光谱、荧光寿命特性及荧光峰值强度随时间变化曲线相差十分明显,反映了癌细胞与正常细胞对血卟啉亲和性有显著的差异;测量结果确认了荧光光谱技术诊断与治疗癌症的可行性,并对发展超短脉冲激光光谱技术早期诊断与治疗癌症具有重要的指导意义和临床应用价值。
OBJECTIVE: To study the ultrafast photodynamic process of photosensitizer hematoporphyrin (HPD) used in the diagnosis and treatment of cancer. Methods: Ultrashort pulse laser spectroscopy and picosecond time-dependent single photon counting system Fluorescence lifetime, Fluorescence lifetime and Fluorescence peak intensity of living cells were compared with those of normal cells. Fluorescence lifetime characteristics were measured at different locations within a single cell. Results: The samples of cancer cells had unique spectral peaks at 645 nm. The fast component of the fluorescence lifetime of the cancer cell sample is about 1200 ps for the slow component of about 150 ps, while the fast component of the normal cell sample is about 2500 ps for the slow component of about 300 ps; the fluorescence peak intensity of the cancer cell sample decays about 10% for 12 hours and the attenuation of the normal cell sample is about 55 %. The fast component of the fluorescence lifetime at 300 ps in the cell is very significant, and the hematoporphyrin concentration in the center is the highest. Conclusion: Fluorescence spectrum, fluorescence lifetime and fluorescence peak intensity of cancer cells and normal cells are obviously different with time curve, reflecting Cancer cells and normal cells have significant differences in hematoporphyrin affinity; the measurement confirms that the fluorescence spectroscopy diagnosis and Feasibility treatment of cancer, and the development of ultrashort pulse laser spectroscopy technique for early diagnosis and treatment of cancer has important significance and clinical application.