利用FCLA特征荧光实现单态氧和超氧的高灵敏检测  

Detection of Singlet Oxygen and Superoxide Anion with the Fluorescence of FCLA

作　　者：魏言春[1] 罗世明[1] 徐未[1] 

机构地区：[1]华南师范大学激光生命科学研究所,暨激光生命科学教育部重点实验室,广东广州510631

出　　处：《光谱学与光谱分析》2008年第11期2633-2635,共3页Spectroscopy and Spectral Analysis

基　　金：国家自然科学基金项目(30470494,30627003,60678050);广东省自然科学基金项目(15012)资助

摘　　要：活性氧(ROS)的检测对生物学和医学研究很多领域方面具有重要意义,如光动力治疗肿瘤的治疗检测、植物应激反应检测、种子活力检测等。实现活性氧中的单态氧、超氧阴离子高灵敏度的检测,是人们追求的目标。研究发现单态氧、超氧的选择性化学发光探针FCLA(fluoresceinyl cypridina luciferin analog)氧化后,在515 nm处荧光会大大增强。利用这个性质并结合非常成熟的荧光检测方法,可以实现活性氧的高灵敏度检测。尤其在微观检测方面,如单细胞,FCLA的化学发光很微弱而难以检测到,但是却可以灵敏地检测到其荧光变化。FCLA荧光检测不仅灵敏度高而且选择性好,其无毒和易于透膜的性质更适于在细胞和生物体上应用。另外如果同时结合FCLA的化学发光,对单态氧和超氧阴离子的检测将更准确、更具实时性。Reactive oxygen species（ROS） plays a major role in various life systems and is critical in understanding the mechanisms of photodynamic therapy（PDT）,seed vigour and plant stress,and so on.Researchers have dedicated much effort for detecting singlet oxygen and suqeroxide anion.FCLA（the fluoresceinyl cypridina luciferin analog） is a singlet oxygen and superoxide anion specific chemiluminescence probe.Typical chemiluminescence signal is relatively weak and its detection at single cell level is rather difficult.In the present study,the fluorescence of FCLA at 515 nm was investigated and was found to increase significantly during the ROS oxidative reaction.With fluorescence imaging technique,ROS can be detected by evaluating the spatial distribution and the dynamic changes in FCLA fluorescence. The technique is proved to have high sensitivity and excellent signal-to-noise ratio compared to direct CL measurement. With minimal cytotoxicity and excellent ROS selectivity,the FCLA fluorescence imaging technique can be a powerful new means in real-time in vivo and in situ single oxygen detection at the single cell level.

关 键 词：FCLA 单态氧 超氧阴离子 荧光 

分 类 号：O657.3[理学—分析化学]