Development of a multispectral multiphoton fluorescence lifetime imaging microscopy system using a streak camera
Development of a multispectral multiphoton fluorescence lifetime imaging microscopy system using a streak camera
复制标题
DOI：
10.1117/12.568736
复制
发表时间：
2005-01
期刊：
影响因子：
--
通讯作者：
J. Qu;Lixin Liu;B. Guo;Zi-yang Lin;T. Hu;Jindong Tian;Shuyan Wang;Jikang Zhang;H. Niu
中科院分区：
文献类型：
--
作者：
J. Qu;Lixin Liu;B. Guo;Zi-yang Lin;T. Hu;Jindong Tian;Shuyan Wang;Jikang Zhang;H. Niu
我们报告了多光谱多光子荧光寿命成像显微镜（MM-FLIM）系统的开发，该系统结合了条纹相机、棱镜分光光度计、飞秒钛宝石激光器和荧光显微镜。该系统用途广泛，具有多光谱功能、高时间 (10ps) 和空间 (0.36μm) 分辨率，可用于使用单个探测器进行 3 维 (3D) (x-y-z) 多光子荧光强度、光谱分辨强度和寿命测量。该系统使用 F-P 标准具和标准荧光染料进行校准，获得的寿命值与文献报道的值非常吻合。初步结果表明，该 MM-FLIM 系统在一个显微镜系统中集成了高时间、空间和光谱分辨率荧光检测。该系统的潜在应用包括多孔成像、组织辨别、细胞内生理学和荧光共振能量转移成像。
We report on the development of a multispectral multiphoton fluorescence lifetime imaging microscopy (MM-FLIM) system that is the combination a streak camera, a prism spectrophotometer, a femtosecond Ti: Sapphire laser and a fluorescence microscope. This system is versatile with multispectral capability, high temporal (10ps) and spatial (0.36μm) resolution and can be used to make 3-dimensional (3D) (x-y-z) multiphoton fluorescence intensity, spectrally resolved intensity and lifetime measurements with a single detector. The system was calibrated with a F-P etalon and a standard fluorescent dye and the lifetime value obtained was in good agreement with the value reported in the literature. Preliminary results suggest that this MM-FLIM system has integrated high temporal, spatial, and spectral resolution fluorescence detection in one microscopy system. Potential applications of this system include multiwell imaging, tissue discrimination, intracellular physiology and fluorescence resonance energy transfer imaging.