摘要:
荧光探针作为研究生物系统必不可少的工具,借助双光子显微成像可以直观便捷地对生物活性化合物或生物功能客体成分进行实时动态三维观测与监控.近十年来发展的双光子激发荧光探针,较单光子荧光探针具有显著的优点,如高分辨率、高清晰度、高灵敏度、无光漂白、无光致毒、定靶激发、高横向与纵向分辨率、低的生物组织吸光系数及低的组织自发荧光干扰等.本文综述了近七年来的双光子阳离子探针、双光子阴离子探针、双光子SO2探针、双光子pH探针、双光子核酸探针、双光子半胱氨酸探针、双光子活性氧探针、双光子磷酸探针、双光子CO探针、双光子Alph-细胞探针、双光子CYP1A酶探针、双光子极性、黏度与温度探针的结构特性及其在生物成像方面的应用.双光子荧光探针已被广泛应用于临床诊断、疾病监测和药物筛选上,这推动了生物化学、医学和生命研究的发展.%Fluorescent probes are essential for the study of biological systems.Using two-photon excited fluorescence microscopy,real-time three-dimensional (3D) dynamic observation and monitoring of biologically active compounds and biological function of object composition can be intuitively and conveniently realized.In recent ten years,the developed two-photon excited fluorescence probes,compared with the one-photon fluorescence probes,have significant advantages,such as high resolution,high clarity,high sensitivity,no photobleaching and no photodamage,fixed target excitation,high transverse and longitudinal resolutions,small absorption coefficient of light in tissue,lower tissue auto-fluorescence,etc.The structures,characteristics and applications in biological imaging of two-photon fluorescence probes for cations,anions,SO2,pH,nucleic acid,cysteine,reactive oxygen species,phosphate,CO,alph-cells,CYP1A enzyme,and polarity,viscosity and temperature in recent seven years are reviewed.Two-photon fluorescent probes have been widely used in clinical diagnosis,disease monitoring and drug screening,which has promoted the development of biochemistry,medicine and life sciences.