荧光寿命取决于荧光分子所处的微环境,通过对样品荧光寿命的测量和成像可以定量获取样品的功能信息.介绍荧光寿命成像(fluorescence lifetime imaging, FLIM)技术原理、实现途径、发展现状及其在生物医学中的应用.提出一种基于皮秒扫描相机、梯度折射率微透镜阵列、棱镜分光光谱仪、飞秒钛宝石激光器和荧光显微物镜的新型五维荧光显微成像技术,该系统具有多光谱分辨功能以及很高的时间和空间分辨率,能够实现三维多光子荧光强度、光谱分辨强度和寿命测量,将在高通量生化分析、组织鉴别、胞内生理学和荧光共振能量转移(fluorescence resonance energy transfer, FRET)等领域获得广泛应用.
Fluorescence lifetime is dependent on the microenvironment of the fluorophore and its measurement and imaging can provide functional information about the sample. The principle, methods, current developments and biomedical applications of fluorescence lifetime imaging (FLIM) were discussed. A novel 5-D fluorescence microscopy that was the combination of a picosecond streak camera, a gradient-index microlens array, a prism spectrophotometer, a femtosecond Ti: Sapphire laser and a fluorescence microscope is presented. The system is versatile with multispectral capability, high temporal and spatial resolution and can be used to make 3-dimensitional multiphoton fluorescence intensity, spectrally resolved intensity and lifetime measurements with a single detector. Potential applications of this system include multiwell imaging, tissue discrimination, intracellular physiology and fluorescence resonance energy transfer imaging.
Journal of Shenzhen University(Science &engineering)