Korczyński Jarosław, Włodarczyk Jakub
Pracownia Mikroskopii Konfokalnej, Instytut Biologii Doświadczalnej im. M. Nenckiego, Warszawa, Poland.
Postepy Biochem. 2009;55(4):434-40.
Fluorescence lifetime imaging microscopy (FLIM) is a powerful tool for producing an image based on the differences in the exponential decay rate of the fluorescence from a fluorescent sample. This technique can provide information, not only concerning the localization of specific fluorophores, but also about the local fluorophore environment. It can be used in scanning confocal, multi-photon microscopes, or in wide-field microscopes and endoscopes. FLIM systems can be implemented both in the frequency domain, using sinusoidally modulated excitation light and in the time domain, using pulsed excitation sources. The power of this technique lies in the fact that the measured fluorescent lifetime of a fluorophore is sensitive to the molecular environment of that fluorophore. Due to this phenomenon FLIM has recently found use in several applications: in the analysis of protein-protein interactions with high spatial and temporal specificity, in ion concentration imaging as well as in measuring of oxygen concentration and in medical applications.
荧光寿命成像显微镜(FLIM)是一种强大的工具,可根据荧光样品荧光指数衰减率的差异生成图像。该技术不仅可以提供有关特定荧光团定位的信息,还能提供有关局部荧光团环境的信息。它可用于扫描共聚焦显微镜、多光子显微镜,或宽视场显微镜和内窥镜。FLIM系统既可以在频域中实现,使用正弦调制激发光,也可以在时域中实现,使用脉冲激发源。该技术的强大之处在于,所测量的荧光团荧光寿命对该荧光团的分子环境敏感。由于这种现象,FLIM最近已应用于多个领域:在具有高空间和时间特异性的蛋白质 - 蛋白质相互作用分析、离子浓度成像、氧浓度测量以及医学应用中。