Development of a multiphoton fluorescence lifetime imaging microscopy system using a streak camera
Development of a multiphoton fluorescence lifetime imaging microscopy system using a streak camera
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DOI：
10.1063/1.1569410
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发表时间：
2003-05-01
影响因子：
1.6
通讯作者：
Herman, B
中科院分区：
文献类型：
--
作者：
Krishnan, RV;Saitoh, H;Herman, B
我们报告的多光子荧光寿命成像系统（FLIM）使用条纹相机的发展和详细的校准。本系统是通用的高空间（类似于0.2妈妈）和时间（类似于50 ps）的分辨率，并允许快速的数据采集和可靠的和可重复的寿命测定。用标准荧光染料校准该系统，得到的寿命值与文献中报道的这些染料的值非常一致。我们还证明了该系统的适用性FLIM研究细胞标本，包括染色的花粉粒和成纤维细胞表达绿色荧光蛋白。所获得的寿命值与其他小组先前报告的这些相同样本的寿命值相匹配。本系统的潜在应用包括细胞内生理学和荧光共振能量转移成像的测量，其在活细胞成像的上下文中讨论。(C)2003年，美国物理学会。
We report the development and detailed calibration of a multiphoton fluorescence lifetime imaging system (FLIM) using a streak camera. The present system is versatile with high spatial (similar to0.2 mum) and temporal (similar to50 ps) resolution and allows rapid data acquisition and reliable and reproducible lifetime determinations. The system was calibrated with standard fluorescent dyes and the lifetime values obtained were in very good agreement with values reported in the literature for these dyes. We also demonstrate the applicability of the system to FLIM studies in cellular specimens including stained pollen grains and fibroblast cells expressing green florescent protein. The lifetime values obtained matched well with those reported earlier by other groups for these same specimens. Potential applications of the present system include the measurement of intracellular physiology and fluorescence resonance energy transfer imaging, which are discussed in the context of live cell imaging. (C) 2003 American Institute of Physics.